Document ID: EPA-HQ-OPP-2002-0233-0018
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2002-09-13T04:00Z

[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Intraperitoneal
Infectivity
Study
/
1
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
3.
3
Reviewer:
Esther
Seto
,
Date
January,
29,
2002
Peer
Review:
Ibrahim
Barsoum,
PhD
Microbial
Pesticides
Branch
Biopesticides
and
Pollution
Prevention
Division
U.
S.
Environmental
Protection
Agency
_______________

STUDY
TYPE:
Intraperitoneal
Infectivity
Study
­
Rat
PMRA
DATA
CODE
M4.3.
3
/
USEPA
OPPTS
885.3200
TEST
MATERIAL
(PURITY):
Sporothrix
flocculosa
(pure
culture)
1.44
x
10
6
CFU/
mL
SYNONYMS:
Pseudozyma
flocculosa
[STF]

CITATION:
Harrington,
Kelly
A.
(June
19,
1997).
"Acute
Intraperitoneal
Infectivity
Testing
of
Sporothrix
flocculosa,
a
Fungal
Pesticide."
IIT
Research
Institute,
Life
Sciences
Department,
Chicago,
Illinois,
USA.
IITRI
Project
No.
L08641.
Study
No.
2.
In­
Life
Dates:
October
17,
1996
­
October
31,
1996.

SPONSOR:
Dr.
Richard
Bélanger
Université
Laval
Québec,
Canada
EXECUTIVE
SUMMARY:
In
an
acute
intraperiteonal
toxicity/
infectivity
study,
groups
of
young
adult
CD
rats
(4/
sex/
scheduled
sacrifice
date)
were
exposed
by
the
intraperitoneal
route
to
an
undiluted
suspension
of
Sporothrix
flocculosa
(TS)
at
a
dose
of
3.
5
x
10
7
cfu/
animal
(in
1.
0
mL).
Animals
were
then
observed
for
up
to
14
days.
An
equal
number
of
young
adult
CD
rats
were
similarly
injected
with
heatkilled
test
substance
(KTS).
An
undosed
naive
control
(NC)
group
consisting
of
4
rats/
sex
was
also
included
in
the
study.
Cage
side
observations
for
clinical
symptoms
was
performed
daily
and
animal
body
weights
and
food
consumption
were
monitored.

No
unscheduled
deaths
occurred.
Designated
animals
from
the
TS
and
KTS
groups
were
sacrificed
on
days
0,
7
and
14
and
gross
necropsies
were
performed.
The
NC
group
of
animals
was
sacrificed
and
necropsied
at
the
end
of
the
14
day
study.
Infectivity
and
clearance
were
assessed
by
quantitatively
recovering
the
MPCA
from
the
blood,
lungs
and
lymph
nodes,
spleen,
kidneys,
liver,
heart,
stomach
and
small
intestine,
peritoneal
fluid,
caecum
and
brain.

No
adverse
clinical
signs
were
observed
at
any
point
of
the
study
in
any
of
the
groups
of
rats.
Body
weight
gain
of
TS­
dosed
male
rats
was
significantly
decreased
while
this
group's
food
consumption
was
significantly
increased
compared
to
NC
animals.
There
was
no
significant
difference
between
KTS­
dosed
and
NC
animals
in
terms
of
body
weight,
body
weight
gain
or
food
consumption.
Upon
necropsy
of
TSand
KTS­
dosed
animals,
white
nodules
and
higher
relative
spleen
weights
were
observed
and
attributed
to
a
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Intraperitoneal
Infectivity
Study
/
2
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
3.
3
normal
immune
response
to
a
foreign
substance.
The
detection
of
S.
flocculosa
in
the
peritoneal
fluid
lavage
of
TS­
dosed
male
rats
was
consistent
with
the
method
of
administration.
Clearance
of
S.
flocculosa
from
all
other
tissues
and
fluids
occurred
by
day
7.
No
test
substance
was
detected
from
any
of
the
organs
of
the
KTS­
dosed
or
NC
animals.

At
the
dose
administered,
S.
flocculosa
is
slightly
toxic
but
not
pathogenic
to
male
and
female
CD
rats
when
introduced
by
the
intraperitoneal
route.

This
acute
intraperitoneal
toxicity/
infectivity
study
is
classified
as
acceptable.

COMPLIANCE:
Signed
and
dated
GLP
and
QA
statements
were
provided.
A
No
Data
Confidentiality
Claims
statement
was
included
but
was
not
signed
or
dated.

I.
MATERIALS
AND
METHODS
A.
MATERIALS:

1.
Test
Material:
Sporothrix
flocculosa
Description:
MPCA
Lot/
Batch
#:
UK34­
35
Purity:

CAS
#:
N/
A
Test
substance
homogeneity
was
determined
by
withdrawing
aliquots
from
three
locations
(top,
centre
and
bottom)
of
a
10
mL
tube
containing
test
substance
in
ASTM
type
1
water.
Samples
from
the
three
locations
were
plated
in
triplicate
on
YM
plates.
Colonies
were
counted
after
incubation
for
three
days
at
25

C.
The
average
plate
count
of
the
triplicate
samples
taken
from
each
location
was
comparable,
indicating
that
the
test
substance
was
homogeneous
throughout
the
volume
of
the
tube.

2.
Sample
Preparation:
Sporothrix
flocculosa
(batch
number
UK34­
35)
was
provided
to
the
test
facility
in
the
form
of
two
petri
dishes
(P1
and
P2)
of
yeast
malt
agar
with
white
mould.
IITRI
then
inoculated
two
malt
agar
plates
(P3
and
P4)
with
a
colony
from
P1.
One
week
prior
to
dosing
in
the
preliminary
challenge
assay
component
of
the
study,
300
mL
of
yeast
malt
broth
was
inoculated
with
test
substance
from
P4
and
incubated
at
room
temperature
for
seven
days.
On
the
day
of
dosing,
the
broth
culture
was
centrifuged
and
the
pellet
was
resuspended
in
5
mL
of
ASTM
Type
1
water.
The
titre
of
the
resuspended
solution
was
determined
to
be
1.
44
x
10
6
cfu/
mL
by
plate
count
(corresponding
to
9.
38
x
10
6
cells/
mL
by
hemacytometer
count).
This
solution
was
used
undiluted
for
dosing.
One
week
before
the
start
of
the
actual
infectivity
study,
300
mL
of
yeast
malt
broth
was
inoculated
with
test
substance
from
P4
and
incubated
at
room
temperature
for
seven
days.
On
the
day
of
dosing,
the
broth
culture
was
centrifuged
and
resuspended
in
7
mL
of
ASTM
Type
1
water.
The
titre
of
the
solution
was
adjusted
to
1
x
10
7
cells/
mL
(by
hemacytometer
count)
and
was
then
divided
into
two
aliquots.
One
aliquot
was
heat­
treated
for
20
minutes
at
approximately
82

C
to
render
the
active
ingredient
non­
viable
and
was
used
as
KTS
(killed
test
substance).
The
second
aliquot
was
used
as
TS
(test
substance).
Both
KTS
and
TS
were
plated
to
verify
titre
and
inactivation,
respectively.
The
titre
of
the
TS
solution
was
3.5
x
10
7
cfu/
mL
and
the
titre
of
the
KTS
solution
was
0
cfu/
mL.
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Intraperitoneal
Infectivity
Study
/
3
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
3.
3
3.
Test
animals:
Species:
Rat
Strain:
Outbred
Sprague­
Dawley
strain,
i.
e.,
CD
Age/
weight
at
dosing:
Preliminary
Challenge:
Males:
49
to
51
days
old,
weights
were
not
reported
Females:
49
to
51
days
old,
weights
were
not
reported
Actual
Study:
Males:
56
to
58
days
old,
244.88
­
288.30
g
Females:
56
to
58
days
old,
184.7
­
213.16
g
Source:
Charles
River
Laboratories,
Portage,
MI,
USA.

Housing:
Rats
were
housed
up
to
two
per
cage
in
polypropylene
cages
with
hardwood
chip
bedding.
Animals
in
the
naive
control
and
killed
test
substance­
dosed
groups
were
housed
in
one
room
while
test
substance­
dosed
groups
were
housed
in
a
second
room.
Diet:
Certified
Purina
Rodent
Chow
5002
(PMI
Feeds,
Inc.,
St.
Louis,
MI,
USA)
was
provided
ad
libitum.
Water:
City
of
Chicago
water
was
provided
ad
libitum.

Environmental
conditions:
Temperature:
Humidity:
Air
changes:
Photoperiod:
18
­
25

C
13
­
70%
Not
Reported
12
hrs
dark
/
12
hrs
light
Acclimation
period:
Preliminary
Challenge:
1
week
Actual
Study:
2
weeks
B.
STUDY
DESIGN
and
METHODS:

1.
In
life
dates
­
Start:
Preliminary
Challenge
­
October
9,
1996
End:
Preliminary
Challenge
­
October
14,
1996
Actual
Study
­October
17,
1996
Actual
Study
­
October
31,
1996
2.
Preliminary
Challenge
Assay
A
preliminary
assay
was
conducted
to
assess
the
toxicity
potential
of
the
test
substance.
The
preliminary
assay
was
initiated
by
administering
the
test
substance
to
three
male
and
three
female
CD
rats
by
intraperitoneal
injection
at
a
dose
of
1.
44
x
10
6
cfu
(9.
38
x
10
6
cells)
in
a
1
mL
dose
per
animal.
Following
treatment,
the
animals
were
observed
for
5
days.
All
animals
survived
and
no
clinical
symptoms
were
observed.

3.
Experimental
Design
Test
animals
were
weighed
and
assigned
to
treatment
groups
such
that
no
animal's
body
weight
varied
from
the
group
mean
body
weight
by
more
than
20%.
Treatment
groups
were
created
according
to
the
treatment
received;
i)
test
substance
(TS),
ii)
killed
test
substance
(KTS),
iii)
no
treatment
or
a
naive
control
group
(NC).
The
number
of
animals
and
the
sacrifice
time
for
each
of
the
test
groups
is
shown
in
Table
1.

TABLE
1.
Treatment,
mortality/
animals
treated
Treatment
Scheduled
Sacrifice
Mortality
(#
dead/
total)

Males
Females
Combined
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Intraperitoneal
Infectivity
Study
/
4
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
3.
3
TS
a
0
0/
4
0/
4
0/
8
KTS
b
0
0/
4
0/
4
0/
8
TS
a
7
0/
4
0/
4
0/
8
KTS
b
7
0/
4
0/
4
0/
8
TS
a
14
0/
4
0/
4
0/
8
KTS
b
14
0/
4
0/
4
0/
8
NC
c
14
0/
4
0/
4
0/
8
a
dosed
with
1
mL
of
TS
containing
3.
5
x
10
7
cfu
of
S.
flocculosa
b
dosed
with
1
mL
of
KTS
containing
the
equivalent
of
3.2
x
10
7
cfu
of
heat­
killed
S.
flocculosa
per
animal
c
naive
control
group
4.
Body
Weights
Animal
body
weights
were
determined
upon
receipt
(random
sample
of
10
animals/
sex),
prior
to
randomization,
at
the
time
of
dosing
(day
0),
and
on
days
7
and
14.

5.
Clinical
Observations
Clinical
observations
were
recorded
daily.

6.
Food
Consumption
Average
daily
food
consumption
per
rat
was
calculated.

7.
Necropsy
Test
animals
were
sacrificed
according
to
Table
1.
Necropsies
consisting
of
body
and
organ
weight
measurements
and
examinations
of
the
external
features,
internal
organs
and
cavities
were
performed.

8.
Microbial
Enumeration
Samples
of
blood,
brain,
caecum,
heart,
kidneys,
liver,
lungs
and
lymph
nodes,
mesenteric
lymph
nodes,
peritoneal
fluid,
spleen,
and
stomach
with
small
intestines
were
aseptically
removed
and
placed
in
sterile
blending
bags
containing
0.
1%
peptone.
The
tissues
were
blended
and
aliquots
were
serially
diluted
in
0.
1
%
peptone
and
plated
onto
duplicate
plates
of
YM
or
MA
(see
study
for
determining
sensitivity
of
detection).
The
plates
were
incubated
for
91
­
94
hours
at
approximately
25

C.
Mean
counts
from
the
duplicate
plates
were
reported
as
viable
cfu/
mL
or
cfu/
g.

9.
Statistics
a)
Body
Weights
and
Food
Consumption
The
mean
and
standard
deviation
in
body
weight
and
average
daily
food
consumption
were
calculated
for
each
treatment
group
by
sex.
The
data
were
log­
transformed
and
statistically
compared
for
treatment
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Intraperitoneal
Infectivity
Study
/
5
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
3.
3
group
by
analysis
of
variance
(ANOVA)
followed,
where
appropriate,
by
Dunnett's
test.
A
p

0.05
was
considered
significant.

b)
Organ
Weights
Group
mean
and
standard
deviation
of
relative
organ
weights
(%
of
total
weight)
were
calculated
for
day
14
data
only.
The
log­
transformed
data
were
statistically
compared
by
treatment
group
by
ANOVA
followed,
where
appropriate,
by
Dunnett's
test.
A
p

0.
05
was
considered
significant.

c)
Microbial
Enumeration
Since
the
test
substance
was
recovered
only
fromTS­
dosed
animals
and
only
on
day
0,
no
statistical
analyses
were
performed.

II.
RESULTS
AND
DISCUSSION:

A.
Mortality
No
deaths
occurred
during
the
study.

B.
Clinical
Observations
No
adverse
clinical
symptoms
were
observed
in
any
of
the
animals
during
the
course
of
the
study.

C.
Body
Weights
and
Body
Weight
Gains
Body
weights
and
body
weight
gains
are
shown
in
Tables
2
and
3,
respectively.
No
significant
differences
between
the
treated
groups
and
control,
with
respect
to
body
weight,
were
observed.
Total
body
weight
gain
and
body
weight
gain
in
male
TS­
dosed
animals
on
day
7,
however,
were
significantly
decreased
compared
to
controls.

Table
2.
Effect
of
Intraperitoneal
Administration
of
Sporothrix
flocculosa
on
Body
Weights
of
Rats
Group
Sex
Parameter
Body
Weight
(g)

Day
0
Day
7
Day
14
TS
M
Mean
StDev
No.
rats
263.65
14.220
12
301.74
19.625
8
309.64
21.810
4
KTS
M
Mean
StDev
No.
rats
265.64
9.9196
12
315.45
10.618
8
332.67
4.296
4
NC
M
Mean
StDev
No.
rats
259.86
12.142
4
310.92
17.580
4
324.54
14.226
4
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Intraperitoneal
Infectivity
Study
/
6
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
3.
3
Group
Sex
Parameter
Body
Weight
(g)

Day
0
Day
7
Day
14
TS
F
Mean
StDev
No.
rats
200.06
8.889
12
220.42
9.548
8
232.59
10.399
4
KTS
F
Mean
StDev
No.
rats
200.57
7.653
12
221.44
12.222
8
234.20
14.497
4
NC
F
Mean
StDev
No.
rats
198.77
7.550
4
218.49
11.458
4
230.61
12.375
4
TABLE
3.
Effect
of
Intraperitoneal
Administration
of
Sporothrix
flocculosa
on
Body
Weight
Gains
of
Rats
Group
Sex
Parameter
Body
Weight
Gain
(g)

Day
0
­7
Day
7
­14
TotalGain
TS
M
Mean
StDev
No.
rats
39.71*
7.081
8
6.58
7.345
4
44.77*
8.942
4
KTS
M
Mean
StDev
No.
rats
45.92
6.353
8
11.04
3.640
4
60.08
4.491
4
NC
M
Mean
StDev
No.
rats
51.06
6.562
4
13.62
10.993
4
64.68
9.753
4
TS
F
Mean
StDev
No.
rats
20.86
6.959
8
8.83
1.749
4
33.52
6.635
4
KTS
F
Mean
StDev
No.
rats
20.37
7.638
8
14.42
8.659
4
31.88
6.818
4
NC
F
Mean
StDev
No.
rats
19.72
16.926
4
12.12
23.703
4
31.84
8.975
4
*significantly
decreased
compared
to
naive
control
(NC)
group
(p

0.05)

D.
Food
Consumption
Average
daily
food
consumption
is
summarized
in
Table
5.
Food
consumption
was
significantly
increased
in
the
male
TS
group
compared
to
the
control.
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Intraperitoneal
Infectivity
Study
/
7
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
3.
3
TABLE
4.
Average
Daily
Food
Consumption
Treatment
Group
Sex
Parameter
Average
Daily
Consumption
(g/
animal)

TS
M
Mean
StDev
No.
rats
26.83*
2.361
8
KTS
M
Mean
StDev
No.
rats
24.97
1.411
8
NC
M
Mean
StDev
No.
rats
23.74
0.809
4
*
significantly
increased
compared
to
naive
control
(NC)
group
(Dunnett's
test,
p

0.05)

TABLE
4
contd.

Treatment
Group
Sex
Average
Daily
Consumption
(g)

TS
F
Mean
StDev
No.
rats
20.22
1.455
8
KTS
F
Mean
StDev
No.
rats
18.77
1.081
8
NC
F
Mean
StDev
No.
rats
18.45
0.745
4
E.
Necropsy
Results
Incidences
of
gross
necropsy
observations
are
summarized
in
Table
5.
No
gross
lesions
were
found
in
any
of
the
NC
animals.
Among
the
animals
which
were
sacrificed
on
day
7,
white
nodules
were
observed
on
the
stomach
of
one
TS
male
rat,
on
the
caecum
of
one
KTS
male
rat,
on
the
liver
of
one
KTS
male
rat,
and
on
the
small
intestine
of
one
TS
female
rat.
White
nodules
were
also
observed
on
the
liver
and
mesenteric
lymph
nodes
of
one
KTS
male
rat
sacrificed
on
day
14.

TABLE
5.
Gross
Necropsy
Observations
TS
KTS
NC
Observation
M
F
M
F
M
F
Number
of
animals
12
12
12
12
4
4
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Intraperitoneal
Infectivity
Study
/
8
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
3.
3
Stomach
­single
white
nodule
1
a
0
0
0
0
0
Caecum
­single
white
nodule
0
0
1
a
0
0
0
Liver
­single
white
nodule
0
0
2
a,
b
0
0
0
Mesenteric
Lymph
Node
­multiple
white
nodules
0
01
b
0
0
0
Small
Intestine
­single
white
nodule
0
1
a
0
0
0
0
No
observed
signs
11
11
9
12
4
4
a
observed
on
day
7
b
observed
on
day
14
F.
Organ
Weights
Relative
organ
weight
data
is
summarized
in
Tables
6
and
7.
Statistical
analysis
of
relative
organ
weights
was
conducted
for
day
14
data
only.
Relative
stomach
and
small
intestine
weights
of
TS­
dosed
female,
TSdosed
male
and
KTS­
dosed
male
rats
were
significantly
decreased
compared
to
naive
controls.
Relative
weights
of
the
lung
and
associated
lymph
nodes
of
KTS­
dosed
female
rats
was
significantly
increased
compared
to
controls.
The
mean
relative
spleen
weight
of
both
KTS­
and
TS­
dosed
female
rats
was
significantly
increased
compared
to
the
NC
group.

Table
6.
Effect
of
Intraperitoneal
Administration
of
S.
flocculosa
on
Relative
Organ/
Tissue
Weights
of
Male
Rats
Relative
Organ
Weight
a
Day
Group
Brain
Heart
Kidneys
Liver
Lung
&
Lymph
Nodes
Mesenteric
Lymph
Nodes
Spleen
Stomach
and
Small
Intestine
0
TS
Mean
b
StDev
0.741
0.0337
0.430
0.0364
0.977
0.0249
5.270
0.5077
0.858
0.0283
0.895
0.3721
0.249
0.0143
5.160
2.7196
KTS
Mean
StDev
0.785
0.0289
0.413
0.0243
1.009
0.0674
5.170
0.1280
0.986
0.3233
0.867
0.2985
0.259
0.0507
4.591
1.2544
7
TS
Mean
StDev
0.684
0.0396
0.460
0.0212
1.001
0.0325
4.475
0.1183
0.605
0.0505
0.519
0.0618
0.259
0.0458
6.821
1.2594
KTS
Mean
StDev
0.676
0.0396
0.406
0.0187
0.931
0.0395
4.876
0.3523
0.611
0.0909
0.667
0.1362
0.271
0.0206
5.795
1.1140
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Intraperitoneal
Infectivity
Study
/
9
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
3.
3
Relative
Organ
Weight
a
Day
Group
Brain
Heart
Kidneys
Liver
Lung
&
Lymph
Nodes
Mesenteric
Lymph
Nodes
Spleen
Stomach
and
Small
Intestine
14
TS
Mean
StDev
0.661
0.0591
0.415
0.0289
0.957
0.0860
3.808
0.4701
0.662
0.0913
0.406
0.1480
0.321
0.2596
2.767*
0.7141
KTS
Mean
StDev
0.639
0.0335
0.456
0.0253
0.868
0.0386
4.128
0.4142
0.622
0.0501
0.589
0.2014
0.220
0.0258
3.291*
0.4613
NC
Mean
StDev
0.630
0.0177
0.435
0.0496
0.956
0.0546
4.268
0.0460
0.543
0.0511
0.507
0.1138
0.218
0.0223
4.676
0.5195
a
Relative
Organ
Weight
=
[Absolute
Organ
Weight
(g)
/
Body
Weight
(g)]
x
100
b
N
=4
*
Statistically
significant
from
NC
(p

0.05)

Table
7.
Effect
of
Intraperitoneal
Administration
of
S.
flocculosa
on
Relative
Organ/
Tissue
Weights
of
Female
Rats.

Relative
Organ
Weight
a
Day
Group
Brain
Heart
Kidneys
Liver
Lung
&
Lymph
Nodes
Mesenteric
Lymph
Nodes
Spleen
Stomach
and
Small
Intestine
0
TS
Mean
b
StDev
0.903
0.0553
0.407
0.0627
1.026
0.0758
4.473
0.5318
0.911
0.3390
0.865
0.5278
0.268
0.0144
4.757
1.2177
KTS
Mean
StDev
0.968
0.0547
0.429
0.0336
0.979
0.0660
4.522
0.1894
1.002
0.2213
1.017
0.3033
0.260
0.0302
5.611
2.4295
7
TS
Mean
StDev
0.917
0.0378
0.515
0.0568
0.929
0.0616
4.392
0.2831
0.647
0.0540
0.595
0.0692
0.254
0.0310
5.577
0.1664
KTS
Mean
StDev
0.875
0.0184
0.444
0.0507
0.859
0.0699
4.392
0.2436
0.617
0.0309
0.523
0.1625
0.250
0.0762
5.275
0.3905
14
TS
Mean
StDev
0.830
0.0433
0.485
0.0768
0.899
0.0804
3.918
0.2160
0.682
0.0297
0.512
0.1119
0.261*
0.0241
2.742*
0.8329
KTS
Mean
StDev
0.806
0.0292
0.497
0.0202
0.892
0.0343
4.691
0.3159
0.827*
0.1596
0.580
0.1438
0.242*
0.0396
5.242
0.7454
NC
Mean
StDev
0.854
0.0665
0.465
0.0503
0.909
0.0512
4.637
0.6254
0.536
0.1133
0.536
0.0486
0.190
0.0164
5.025
0.8028
a
Relative
Organ
Weight
=
[Absolute
Organ
Weight
(g)
/
Body
Weight
(g)]
x
100
b
N
=3
*
Statistically
significant
from
NC
(p

0.05)

G.
Test
Substance
Enumeration
Tables
8
and
9
summarize
the
results
of
microbial
enumeration
from
various
organs,
blood
and
peritoneal
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Intraperitoneal
Infectivity
Study
/
10
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
3.
3
fluid.
Sporothrix
flocculosa
was
recovered
from
the
caecum,
kidneys,
liver,
lungs
and
associated
lymph
nodes,
mesenteric
lymph
nodes,
spleen
and
stomach
and
small
intestines
of
both
male
and
female
TS­
dosed
rats.
Sporothrix
flocculosa
was
also
recovered
from
the
peritoneal
fluid
of
TS­
dosed
male
rats
and
from
the
blood
of
TS­
dosed
female
rats.
The
test
substance
was
only
recovered
from
animals
sacrificed
on
day
0
(post­
dosing),
indicating
clearance
within
7
days.
No
test
substance
was
recovered
from
the
KTS­
dosed
or
NC
groups
of
animals.
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Intraperitoneal
Infectivity
Study
/
11
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
3.
3
TABLE
8.
Recovery
of
Sporothrix
flocculosa
Following
Intraperitoneal
Administration
from
Selected
Tissues/
Fluids
of
Male
Rats
Log
[(
cfu/
g
of
tissue)
+
1]
or
Log
[(
cfu/
mL
of
blood
or
peritoneal
fluid)
+
1]

Group
Day
Blood
Brain
Caecum
Heart
Kidney
s
Live
r
Lungs
&

Lymph
Nodes
Mesenteric
Lymph
Nodes
Peritoneal
Fluid
Spleen
Stomach
&

Small
Intestine
TS
0
Mean
SD
GeoMn
b
N
BDL
a
BDL
BDL
4
BDL
BDL
BDL
4
2.791
1.872
6.16
x
10
2
4
BDL
BDL
BDL
4
2.134
1.502
1.35
x
10
2
4
4.031
0.729
1.07
x
10
4
4
1.721
2.039
5.16
x
10
1
4
4.844
1.188
6.99
x
10
4
4
2.565
2.962
3.66
x
10
2
4
3.134
2.192
1.36
x
10
3
4
6.049
0.358
1.15
x
10
6
4
KTS
0
Mean
SD
GeoMn
N
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
TS
7
Mean
SD
GeoMn
N
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
KTS
7
Mean
SD
GeoMn
N
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
TS
14
Mean
SD
GeoMn
N
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
KTS
14
Mean
SD
GeoMn
N
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
NC
14
Mean
SD
GeoMn
N
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
a
BDL
­
below
detection
limit
b
GeoMn
­
geometric
mean
of
CFU/
tissue
or
mL
of
blood
or
peritoneal
fluid
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Intraperitoneal
Infectivity
Study
/
12
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
3.
3
TABLE
9.
Recovery
of
Sporothrix
flocculosa
Following
Intraperitoneal
Administration
from
Selected
Tissues/
Fluids
of
Female
Rats
Log
[(
CFU/
g
of
tissue)
+
1]
or
Log
[(
CFU/
mL
of
blood
or
peritoneal
fluid)
+
1]

Group
Day
Blood
Brain
Caecum
Heart
Kidney
s
Live
r
Lungs
&

Lymph
Nodes
Mesenteric
Lymph
Nodes
Peritoneal
Fluid
Spleen
Stomach
&

Small
Intestine
TS
0
Mean
SD
GeoMn
b
N
0.644
1.288
4.41
x
10
0
4
BDL
BDL
BDL
4
5.421
1.200
2.64x
10
5
4
BDL
BDL
BDL
4
0.734
1.467
5.42
x
10
0
4
3.392
2.461
2.47
x
10
3
4
1.322
1.527
2.00
x
10
1
4
3.698
2.748
4.99
x
10
3
4
BDL
BDL
BDL
4
1.977
2.283
9.38
x
10
1
4
4.477
1.257
3.00
x
10
4
4
KTS
0
Mean
SD
GeoMn
N
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
TS
7
Mean
SD
GeoMn
N
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
KTS
7
Mean
SD
GeoMn
N
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
TS
14
Mean
SD
GeoMn
N
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
KTS
14
Mean
SD
GeoMn
N
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
NC
14
Mean
SD
GeoMn
N
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
BDL
BDL
BDL
4
a
BDL
­
below
detection
limit
b
GeoMn
­
geometric
mean
of
CFU/
tissue
or
mL
of
blood
or
peritoneal
fluid
H.
Reviewer's
Conclusions
This
intraperitoneal
infectivity
study
is
classified
as
acceptable.
Sporothrix
flocculosa
was
considered
to
be
non­
pathogenic
but
slightly
toxic,
at
the
dose
administered
via
the
intraperitoneal
route,
due
to
decreased
body
weight
gain
coupled
with
increased
food
consumption
in
TS­
dosed
male
rats.
The
nodules
observed
at
necropsy
in
both
TS­
and
KTS­
dosed
rats
and
the
higher
relative
spleen
weights
were
the
result
of
the
body's
normal
immunological
response
to
a
foreign
substance.
Changes
in
the
relative
weight
of
stomach
and
small
intestines
of
TS­
and
KTS­
dosed
animals
were
likely
not
biologically
significant
as
evidenced
by
the
variability
in
the
data.
The
detection
of
S.
flocculosa
in
the
peritoneal
fluid
lavage
of
TS­
dosed
male
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Intraperitoneal
Infectivity
Study
/
13
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
3.
3
rats
was
consistent
with
the
method
of
administration.
Clearance
of
S.
flocculosa
from
all
other
tissues
and
fluidsoccurred
by
day7.

I.
Deficiencies
The
number
of
air
changes
per
hour
was
not
reported.
However,
given
the
data
presented,
this
omission
is
considered
to
have
no
impact
on
the
interpretation
of
the
study
results.
Also,
the
relative
humidity
recorded
in
the
animal
room
was
outside
the
preferred
range,
30­
70%.
This
occurrence
is
considered
to
have
no
adverse
effects
on
the
outcome
of
this
study.