Document ID: EPA-HQ-OPP-2005-0017-0004
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2005-09-01T04:00Z

Page
1
of
48
UNITED
STATES
ENVIRONMENTAL
PROTECTION
AGENCY
WASHINGTON,
D.
C.
20460
OFFICE
OF
PREVENTION,
PESTICIDES
AND
TOXIC
SUBSTANCES
MEMORANDUM
DATE:
8/
17/
2005
SUBJECT:
Kasugamycin.
Human
Health
Risk
Assessment
for
Proposed
Food
Uses
of
the
Fungicide
Kasugamycin
on
Imported
Fruiting
Vegetables
(
Group
8).

Petition
Number:
3E6579.
PC
Code
(
Chemical
Number):
230001
DP
Barcode:
D301735
Regulatory
Citation:
40CFR
§
180.???
(
Not
Yet
Established)
EPA
Registration
Number:
(
Not
Registered)
Trade
Name:
Kasumin
®
2L
Chemical
Class:
Aminoglycoside
Antibiotic
Fungicide
Regulatory
Action:
Establishment
of
Permanent
Tolerances
Risk
Assessment
Type:
Single
Chemical,
No
Aggregate
FROM:
William
T.
Drew,
Chemist/
Risk
Assessor
Kelly
M.
Schumacher,
Toxicologist
Douglas
A.
Dotson,
Dietary
Exposure
Assessor
Registration
Action
Branch
2
Health
Effects
Division
(
7509C)

THROUGH:
Richard
A.
Loranger,
Branch
Senior
Scientist
Registration
Action
Branch
2
Health
Effects
Division
(
7509C)

TO:
Mary
Waller/
Lana
Coppolino,
RM
Team
21
Fungicide
Branch
Registration
Division
(
7505C)
Page
2
of
48
Table
of
Contents
1.0
Executive
Summary
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4
2.0
Ingredient
Profile
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6
2.1
Summary
of
Proposed
Uses
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6
2.2
Structure
and
Nomenclature
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7
2.3
Physical
and
Chemical
Properties
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8
3.0
Metabolism
Assessment
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8
3.1
Comparative
Metabolic
Profile
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8
3.2
Nature
of
the
Residue
in
Foods
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9
3.2.1.
Description
of
Primary
Crop
Metabolism
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9
3.2.2
Description
of
Livestock
Metabolism
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9
3.2.3
Description
of
Rotational
Crop
Metabolism
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9
3.3
Environmental
Degradation
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9
3.4
Tabular
Summary
of
Metabolites
and
Degradates
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10
3.5
Toxicity
Profile
of
Major
Metabolites
and
Degradates
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11
3.6
Summary
of
Residues
for
Tolerance
Expression
and
Risk
Assessment
.
.
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11
4.0
Hazard
Characterization/
Assessment
.
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12
4.1
Hazard
Characterization
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12
4.2
FQPA
Hazard
Considerations
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19
4.2.1
Adequacy
of
the
Toxicity
Data
Base
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19
4.2.2
Evidence
of
Neurotoxicity
.
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19
4.2.3
Developmental
Toxicity
Studies
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20
4.2.3.1
Developmental
Toxicity
Study
in
Rats
.
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20
4.2.3.2
Developmental
Toxicity
Study
in
Rabbits
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21
4.2.4
Reproductive
Toxicity
Study
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21
4.2.5
Additional
Information
from
Literature
Sources
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22
4.2.6
Pre­
and/
or
Post­
Natal
Toxicity
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22
4.2.6.1
Determination
of
Susceptibility
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22
4.2.6.2
Degree
of
Concern
Analysis
and
Residual
Uncertainties
for
Pre­
and/
or
Post­
natal
Susceptibility
.
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23
4.3
Recommendation
for
a
Developmental
Neurotoxicity
Study
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23
4.3.1
Evidence
that
Supports
Requiring
a
Developmental
Neurotoxicity
Study
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23
4.3.2
Evidence
that
Supports
not
Requiring
for
a
Developmental
Neurotoxicity
Study
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23
4.4
Hazard
Identification
and
Toxicity
Endpoint
Selection
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23
4.4.1
Acute
Reference
Dose
(
aRfD)
­
Females
(
13
to
49
Years
of
Age)
.
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23
4.4.2
Acute
Reference
Dose
(
aRfD)
­
General
Population
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24
4.4.3
Chronic
Reference
Dose
(
cRfD)
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24
4.4.4
Incidental
Oral
Exposure
(
Short
and
Intermediate
Term)
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26
4.4.5
Dermal
Absorption
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26
4.4.6
Dermal
Exposure
(
Short,
Intermediate
and
Long
Term)
.
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26
Page
3
of
48
4.4.7
Inhalation
Exposure
(
Short,
Intermediate
and
Long
Term)
.
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26
4.4.8
Margins
of
Exposure
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27
4.4.9
Recommendation
for
Aggregate
Exposure
Risk
Assessments
.
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27
4.4.10
Classification
of
Carcinogenic
Potential
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27
4.5
Special
FQPA
Safety
Factor
.
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28
4.6
Endocrine
Disruption
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28
5.0
Public
Health
Data
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29
6.0
Exposure
Characterization/
Assessment
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29
6.1
Dietary
Exposure/
Risk
Pathway
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29
6.1.1
Residue
Profile
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29
6.1.2
Acute
and
Chronic
Dietary
Exposure
and
Risk
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.
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30
6.2
Water
Exposure/
Risk
Pathway
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31
6.3
Residential
(
Non­
Occupational)
Exposure/
Risk
Pathway
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31
7.0
Aggregate
Risk
Assessments
and
Risk
Characterization
.
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31
8.0
Cumulative
Risk
Characterization/
Assessment
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31
9.0
Occupational
Exposure/
Risk
Pathway
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31
10.0
Data
Needs
and
Label
Requirements
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31
10.1
Toxicology
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31
10.2
Residue
Chemistry
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32
References:
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32
Appendices
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33
Page
4
of
48
1.0
Executive
Summary
Arvesta
Corporation,
as
an
agent
for
Hokko
Chemical
Industry
Company,
Limited
(
Japan),
has
proposed
(
in
PP#
3E6579)
the
establishment
of
permanent
tolerances
for
residues
of
the
fungicide
kasugamycin,
with
CAS
Registry
Number
6980­
18­
3
and
CAS
Name
3­
O­[
2­
amino­
4­[(
carboxyiminomethyl)
amino]­
2,3,4,6­
tetradeoxy­
 ­
D­
arabino­
hexopyranosyl]­
D­
chiro­
inositol,
in/
on
the
agricultural
commodities
listed
below.
The
proposal
is
for
tolerances
on
imported
commodities
without
a
US
registration.
Fruiting
vegetables
(
Crop
Group
8)
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
0.04
ppm
Tomato
juice
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
0.06
ppm
Tomato
puree
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
0.06
ppm
Tomato
paste
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
0.25
ppm
The
nature
of
the
residue
in
fruiting
vegetables
is
adequately
understood,
based
upon
an
acceptable
study
of
kasugamycin
metabolism
in
tomato
fruit
and
foliage.
Results
of
the
submitted
tomato
metabolism
study
indicate
that
the
major
metabolic
pathway
in
tomato
involves
conjugation
of
the
parent
compound,
conversion
to
kasugamycinic
acid,
and
subsequent
conjugation
of
kasugamycinic
acid.
Conversion
of
kasugamycin
to
2­
N­
acetyl
kasugamycin
and
kasuganobiosamine
is
thought
to
be
a
minor
metabolic
route.
The
residue
of
concern
(
ROC)
in
fruiting
vegetables
(
for
both
tolerance
setting
and
risk
assessment
purposes)
is
the
parent
compound,
kasugamycin
per
se
(
Kasugamycin
RAD
Team
decision,
D301735,
meeting
of
2/
23/
2005).
There
are
currently
no
established
Codex,
Canadian,
or
Mexican
MRLs
for
kasugamycin.
The
toxicology
database
is
adequate
for
purposes
of
risk
assessment.
Kasugamycin
exhibits
low
acute
toxicity.
The
major
effects
observed
across
species
in
multiple
dose
studies
were
decreased
body
weights
and
body
weight
gains.
Kasugamycin
is
not
mutagenic
and
no
oncogenic
potential
was
noted
in
the
mouse
oncogenicity
or
in
the
rat
combined
chronic/
carcinogenicity
studies.
Based
on
the
overall
weight
of
the
evidence,
kasugamycin
is
classified
as
"
not
likely
to
be
carcinogenic
to
humans."
There
was
no
evidence
that
exposure
to
kasugamycin
results
in
neurotoxicity.
The
database
is
complete
with
respect
to
pre­
and
postnatal
toxicity
and
shows
no
evidence
of
increased
qualitative
or
quantitative
susceptibility
in
the
offspring.
Based
on
the
hazard
and
exposure
data,
HED
recommends
that
the
special
FQPA
safety
factor
be
reduced
to
1X
and
that
no
database
uncertainty
factor
is
needed
because
there
are
no/
low
concerns
and
no
residual
uncertainties
with
regard
to
pre­
and/
or
post­
natal
toxicity.
Neither
dose
nor
endpoint
was
selected
for
acute
dietary
exposure
in
the
general
population
(
or
in
any
of
the
population
subgroups)
because
an
acute
toxic
endpoint
was
not
identified
in
the
submitted
studies.
The
chronic
dietary
endpoint
is
based
on
increased
testicular
softening
and
atrophy
observed
in
a
combined
chronic
toxicity/
carcinogenicity
study
in
rats.
The
NOAEL
for
this
study
is
11.3
mg/
kg/
day,
resulting
in
a
chronic
reference
dose
(
cRfD)
and
chronic
population
adjusted
dose
(
cPAD)
of
0.113
mg/
kg/
day.
The
chronic
dietary
assessment
was
conducted
using
DEEM­
FCID
and
is
very
conservative
in
that
100%
crop
treated
and
tolerance­
level
residues
were
used
in
all
commodities.
Exposure
estimates
include
the
ROC
for
kasugamycin
originating
from
food
only.
With
no
proposed
US
registration,
there
is
no
expectation
that
kasugamycin
residues
would
occur
in
surface
or
ground
water
sources
of
drinking
water.
Since
kasugamycin
is
proposed
for
use
only
on
imported
fruiting
vegetable
commodities,
the
sole
anticipated
exposure
route
for
the
US
population
is
via
dietary
(
food
only)
exposure.
The
Tier
1
analysis
indicates
that
chronic
dietary
exposures
to
kasugamycin
are
below
HED's
level
of
concern
(
100%
of
the
cPAD).
For
chronic
Page
5
of
48
dietary
(
food
only)
exposure
to
kasugamycin,
the
most
highly
exposed
population
subgroup
is
Children
(
1
to
2
years
of
age),
which
utilizes
less
than
1%
of
the
cPAD.
Chronic
dietary
risk
to
all
other
subgroups
is
less
than
that
of
Children
(
1­
2).
With
no
proposed
US
registration,
kasugamycin
is
not
intended
for
use
in
public,
residential,
or
occupational
settings;
also,
there
is
no
expectation
that
exposure
to
kasugamycin
residues
would
occur
via
water
consumption.
Ergo,
risk
assessments
for
water,
residential,
aggregate,
and
occupational
exposures
were
not
performed.
Based
on
the
submitted
crop
field
trial
data,
the
following
use
pattern
would
be
supported
for
fruiting
vegetables:
application
of
the
Kasumin
®
2L
formulation
as
a
foliar
broadcast
spray
in
a
minimum
spray
volume
of
30
gallons
of
water
per
acre
(
GPA)
using
ground
equipment,
with
no
spray
adjuvants,
a
minimum
re­
treatment
interval
(
RTI)
of
3
days,
and
a
pre­
harvest
interval
(
PHI)
of
1
day.
The
submitted
field
trial
data
for
peppers
and
tomatoes
are
adequate
to
support
the
proposed
uses
on
imported
fruiting
vegetables.
No
residues
were
detected
above
the
limit
of
quantitation
(
LOQ);
therefore
the
proposed
and
recommended
tolerances
are
based
on
the
LOQ.
Processing
studies
on
tomato,
for
the
processed
commodities
tomato
puree
and
tomato
paste,
have
been
waived.
The
proposed
residue
analytical
method
is
adequate
for
data
collection.
The
method
was
submitted
to
the
Analytical
Chemistry
Laboratory
of
the
Biological
and
Economic
Analysis
Division
(
ACL/
BEAD)
for
a
petition
method
validation
(
PMV)
trial
(
PMV
Request
Memo,
D313672,
William
T.
Drew,
3/
1/
2005)
to
determine
its
suitability
for
tolerance
enforcement.
The
method
subsequently
failed
the
PMV
trial
(
PMV
Results
Memo,
D313673,
Patricia
G.
Schermerhorn,
7/
13/
2005).
The
method
should
be
modified
as
recommended
by
ACL.
The
fact
that
an
adequate
analytical
method
for
tolerance
enforcement
is
unavailable
does
not
impact
this
risk
assessment
regarding
risk
characterization.
HED
has
examined
the
residue
chemistry
database
for
kasugamycin
and
recommends
against
the
establishment
of
tolerances
for
residues
of
this
fungicide
in/
on
imported
fruiting
vegetables
until
the
data
deficiencies
listed
below
(
OPPTS
Residue
Chemistry
Test
Guideline
860.1340)
have
been
resolved.
860.1340
Residue
Analytical
Method
(
Plant
Commodities)
(
1)
The
proposed
enforcement
method
should
be
modified
to
include
a
confirmatory
analysis
method;
alternatively,
the
petitioner
may
submit
an
interference
study
with
all
pesticides
for
which
tolerances
on
tomatoes
and/
or
peppers
have
been
established.
(
2)
The
method
has
failed
a
PMV
trial
(
PMV
Results
Memo,
D313673,
Patricia
G.
Schermerhorn,
7/
13/
2005),
and
should
be
modified
to
include
the
revisions
recommended
by
ACL.
860.1550
Proposed
Tolerances
The
available
crop
field
trial
data
support
a
tolerance
for
residues
of
kasugamycin
per
se
in/
on
imported
fruiting
vegetable
commodities
(
Crop
Group
8)
at
0.04
ppm,
the
method's
LOQ.
The
exaggerated­
rate
tomato
field
trial
data
(
reflecting
a
5X
application
rate)
indicate
that
separate
tolerances
for
tomato
juice,
tomato
puree
and
tomato
paste
are
not
required.
The
proposed
tolerances
should
be
revised
to
reflect
the
tolerance
as
recommended
by
HED
and
the
correct
commodity
definition
as
specified
in
Table
1.0,
below.
The
petitioner
should
submit
a
revised
Section
F
for
PP#
3E6579
to
reflect
these
changes.
Page
6
of
48
TABLE
1.0
Tolerance
Summary
for
Kasugamycin.

Commodity
Proposed
Tolerance
(
ppm)
Recommended
Tolerance
(
ppm)
Comments/
Correct
Commodity
Definition
Fruiting
Vegetables
(
Crop
Group
8)
0.04
0.04
Vegetable,
fruiting,
group
8
Tomato
Juice
0.06
None
Residues
are
not
expected
to
exceed
the
tolerance
on
the
raw
agricultural
commodity
(
RAC).
Tomato
Puree
0.06
Tomato
Paste
0.25
2.0
Ingredient
Profile
Kasugamycin
is
a
low­
use­
rate
wide­
spectrum
aminoglycoside
antibiotic
fungicide
produced
from
Streptomyces
kasugaensis
that
is
intended
for
control
of
bacterial
and
fungal
diseases
on
pepper
and
tomato.
Kasugamycin
has
preventative,
curative,
and
systemic
activity
at
low
application
rates.
It
is
currently
used
in
over
20
countries
on
various
crops
including
tomato,
paprika,
eggplant,
rice,
and
potato.
Kasugamycin
has
a
mode
of
action
different
from
that
of
the
other
aminoglycoside
antibiotics
such
as
streptomycin.
Because
kasugamycin
is
active
only
against
phytopathogenic
fungi
and
bacteria,
it
has
never
been
employed
as
a
human
or
veterinaryuse
antibiotic.

2.1
Summary
of
Proposed
Uses
The
petitioner
has
indicated
that
Arvesta
de
Mexico
is
seeking
registration
of
Kasumin
®
2L,
a
liquid
formulation
comprised
of
2%
kasugamycin
(
by
weight)
as
the
active
ingredient
(
ai),
for
use
on
rice,
potato,
pepper,
and
tomato
in
Mexico.
The
product
is
formulated
from
kasugamycin
hydrochloride
hydrate
(
2.3%)
and
contains
2%
kasugamycin
(
0.1667
lb
ai/
gal)
as
the
free
base.
Kasugamycin
is
also
formulated
as
a
co­
active
ingredient
(
at
5%)
along
with
copper
oxychloride
(
at
45%,
expressed
as
copper)
in
a
wettable
powder
(
WP),
designated
Kasumin
Cobre
®
.
Additionally,
the
Kasumin
®
formulation
is
a
WP
containing
8%
kasugamycin.
Only
tomatoes
and
peppers
will
be
imported
into
the
US
from
Mexico.
The
current
petition
proposes
use
of
kasugamycin
in
Mexico,
where
the
principal
target
organisms
are
bacteria
rot
(
Erwinia
atroseptica)
and
leaf
mold
(
Cladosporium
fulvum)
on
tomato,
and
bacteria
spot
(
Xanthomonas
campestris,
pv
vesicatoria)
on
both
tomato
and
pepper.
The
petitioner
is
proposing
application
of
kasugamycin
to
peppers
and
tomatoes
as
a
foliar
broadcast
spray
with
up
to
3
applications
at
a
maximum
single
application
rate
of
0.018
pound
ai
per
acre
(
lb
ai/
A),
a
maximum
seasonal
application
rate
of
0.054
lb
ai/
A,
and
a
1­
day
PHI.
Proposed
application
timing
(
as
related
to
plant
growth
stage),
application
equipment
type,
retreatment
intervals
(
RTIs),
spray
volumes,
and
whether
tank
mix
adjuvants
would
be
proposed
were
not
specified.
Based
on
the
submitted
field
trial
data,
the
following
use
pattern
would
be
supported
for
fruiting
vegetables:
application
of
the
Kasumin
®
2L
formulation
as
a
foliar
broadcast
spray
in
a
minimum
spray
volume
of
30
GPA
using
ground
equipment,
with
no
spray
adjuvants,
a
minimum
RTI
of
3
days,
and
a
PHI
of
1
day.
Page
7
of
48
O
O
OH
OH
OH
OH
O
H
C
H
3
NH
NH
O
H
O
NH
2
[

HCl

H2O]
TABLE
2.1
Summary
of
Directions
for
Use
of
Kasugamycin.

Trade
Name
Formulation
[
EPA
Reg.
Number]
Application
Type/
Timing
and
Equipment
Application
Rate
(
lb
ai/
A)
Maximum
Number
of
Applications
per
Season
Maximum
Seasonal
Application
Rate
(
lb
ai/
A)
PHI
(
Days)
Use
Directions
and
Limitations
Proposed
Use
on
Fruiting
Vegetables
(
Group
8)
Imported
from
Mexico
Kasumin
®
2L
[
None]
Foliar
broadcast/
none
specified
0.018
3
0.054
1
None
specified
2.2
Structure
and
Nomenclature
TABLE
2.2
Test
Compound
Nomenclature.

Chemical
Structure
Empirical
Formula
C14H25N3O9
[
C14H25N3O9°
HCl°
H2O]

Common
Name
Kasugamycin
[
Kasugamycin
hydrochloride
hydrate]

Company
Experimental
Name
TM­
416
IUPAC
Name
(
Kasugamycin)
1L­
1,3,4/
2,5,6­
1­
deoxy­
2,3,4,5,6­
pentahydroxycyclohexyl­
2­
amino­
2,3,4,6­
tetradeoxy­
4­(
 ­
iminoglycino)­
 ­
D­
arabino­
hexopyranoside
or
[
5­
amino­
2­
methyl­
6­(
2,3,4,5,6­
pentahydroxycyclohexyloxy)
tetrahydropyran­
3­
yl]
amino­
 ­
iminoacetic
acid
CAS
Name
(
Kasugamycin)
3­
O­[
2­
amino­
4­[(
carboxyiminomethyl)
amino]­
2,3,4,6­
tetradeoxy­
 ­
D­
arabinohexopyranosyl
D­
chiro­
inositol
CAS
Registry
Number
6980­
18­
3
[
19408­
46­
9]

End­
Use
Product
(
EUP)
Kasumin
®
2L
Chemical
Class
Aminoglycoside
antibiotic
fungicide
Known
Impurities
of
Concern
None
Page
8
of
48
2.3
Physical
and
Chemical
Properties
TABLE
2.3
Physicochemical
Properties
of
the
Technical
Grade
Compound
(
Kasugamycin
Hydrochloride
Hydrate).

Parameter
Value
Reference
Molecular
Weight
433.8
PP#
3E6579
administrative
materials
(
MRIDs
#
45910004
and
­
05)
Melting
Point/
Range
202­
230

C
(
decomposing)

pH
4.35
at
24.5

C
(
1%
wt/
vol
solution)

Density
0.43
g/
mL
at
24.5

C
Water
Solubility
g/
100
mL
pH
5
20.7
pH
7
22.8
pH
9
43.8
Solvent
Solubility
g/
100
mL
Methanol
0.744
Hexane
<
1
x
10­
5
Acetonitrile
<
1
x
10­
5
Methylene
chloride
<
1
x
10­
5
Vapor
Pressure
<
0.013
mPa
at
25

C
Dissociation
Constant
(
pKa)
pKa1
=
3.23
pKa2
=
7.73
pKa3
=
11.0
Octanol/
Water
Partition
Coefficient
(
Log
[
KOW])
<
1.96
at
23

C
and
pH
5
UV/
Visible
Absorption
Spectrum
Not
available
3.0
Metabolism
Assessment
3.1
Comparative
Metabolic
Profile
In
the
rat
metabolism
study,
the
mean
radioactivity
recovery
168
hours
after
exposure
ranged
from
91
to
97%,
with
the
majority
of
the
dose
recovered
within
48
hours
in
the
feces
(
81.9­
93.9%)
and
urine
(
1.26­
3.07%).
The
maximum
concentration
found
in
the
plasma
of
both
males
and
females
occurred
approximately
one
hour
after
the
administration
of
a
single
low
or
high
dose.
Between
one
and
six
hours
after
a
single
low
or
high
dose,
more
kasugamycin
accumulated
in
the
kidneys,
urinary
bladder,
and
lymph
nodes
than
in
the
blood,
but
after
168
hours,
little
or
no
kasugamycin
was
found
in
these
tissues.
The
absorption
and
metabolism
of
kasugamycin
in
rats
was
limited
(
less
than
5%
of
the
dose)
and
was
not
affected
by
sex,
dose
level,
or
duration
of
dosing.
The
parent
compound
was
the
major
component
identified
in
the
urine,
feces,
liver,
kidney,
and
plasma.
Minor
amounts
(
less
than
1%
of
the
dose)
of
the
Page
9
of
48
O
O
OH
OH
OH
OH
O
H
C
H
3
NH
NH
O
H
O
NH
2
Cl
H
O
H
2
.
.

*
*
*
*
*

*
metabolite
kasuganobiosamine
were
identified
in
urine,
liver,
kidney,
and
plasma,
but
none
was
detected
in
the
feces.
Elimination
occurred
primarily
in
the
feces
(
88
to
95%),
suggesting
low
absorption;
kasugamycin
was
not
excreted
in
the
bile
(
enterohepatic
circulation
did
not
occur).
In
the
tomato
metabolism
study,
the
metabolite
profile
was
similar
for
tomato
fruit
and
foliage.
Based
on
the
results
of
the
tomato
metabolism
study
the
petitioner
proposes
that
the
major
metabolic
pathway
of
kasugamycin
in
plants
involves
conjugation
of
the
parent
compound,
conversion
to
kasugamycinic
acid,
and
subsequent
conjugation
of
kasugamycinic
acid.
Conversion
of
kasugamycin
to
2­
N­
acetyl
kasugamycin
and
kasuganobiosamine
was
thought
to
be
a
minor
metabolic
route.
Parent
compound
(
kasugamycin
per
se)
was
the
major
identified
component
in
all
samples
from
all
harvest
intervals.

3.2
Nature
of
the
Residue
in
Foods
3.2.1.
Description
of
Primary
Crop
Metabolism
Arvesta
Corporation
has
submitted
a
study
investigating
the
metabolism
of
[
14C]­
kasugamycin
in
tomatoes.
The
study
is
adequate
for
delineating
the
major
residues.
Based
on
the
results
of
the
tomato
metabolism
study
the
petitioner
proposes
that
the
major
metabolic
pathway
of
kasugamycin
in
tomatoes
involves
conjugation
of
the
parent
compound,
conversion
to
kasugamycinic
acid,
and
subsequent
conjugation
of
kasugamycinic
acid.
Conversion
of
kasugamycin
to
2­
N­
acetyl
kasugamycin
and
kasuganobiosamine
was
thought
to
be
a
minor
metabolic
route.

TABLE
3.2.1
Characteristics
of
Test
Material
Used
in
the
Metabolism
Study.

Chemical
Structure
Common
Name
[
14C]­
Kasugamycin
hydrochloride
hydrate
Radiolabel
Position
(
*
)
All
carbons,
including
the
methyl
substituent,
in
the
2,3,4,6­
tetradeoxy­
 ­
D­
arabino­
hexopyranosyl
ring
3.2.2
Description
of
Livestock
Metabolism
There
are
no
significant
livestock
feed
items
associated
with
this
petition;
therefore,
the
nature
of
the
residue
in
livestock
does
not
need
to
be
addressed.

3.2.3
Description
of
Rotational
Crop
Metabolism
Per
the
NAFTA
Guidance
Document
on
Data
Requirements
for
Tolerances
on
Imported
Commodities
(
dated
April
2003),
rotational
crop
data
are
not
required
to
support
the
proposed
Page
10
of
48
O
O
OH
OH
OH
OH
O
H
C
H
3
NH
NH
O
H
O
NH
2
O
O
OH
OH
OH
OH
O
H
C
H
3
NH
O
O
H
O
NH
2
tolerances
on
imported
fruiting
vegetables.

3.3
Environmental
Degradation
Since
kasugamycin
is
proposed
for
use
only
on
imported
fruiting
vegetable
commodities,
with
neither
existing
nor
proposed
US
registration,
there
is
no
expectation
that
kasugamycin
residues
would
occur
in
surface
or
ground
water.

3.4
Tabular
Summary
of
Metabolites
and
Degradates
TABLE
3.4
Tabular
Summary
of
Kasugamycin
Metabolites
and
Degradates.

Common
Name
[
Code]
%
Total
Radioactive
Residues
Chemical
Name
Chemical
Structure
Kasugamycin,
Parent
Compound
[
TM­
416]

Rat
Essentially
100%
TRR
Tomato
(
Fruit)
90­
94%
TRR
(
2­
hour,
1­
day)
69%
TRR
(
7­
day)
55­
60%
TRR
(
14­,
21­,
28­
day)
Tomato
(
Foliage)
75­
84%
TRR
(
2­
hour,
1­
day)
70%
TRR
(
7­
day)
52­
57%
TRR
(
14­,
21­,
28­
day)
[
5­
amino­
2­
methyl­
6­
(
2,3,4,5,6­
pentahydroxycyclohexyloxy)
tetrahydro­
pyran­
3­
yl]
amino­
 ­
iminoacetic
acid
Kasugamycinic
acid
[
KA­
2]

Tomato
(
Fruit)
10%
TRR
(
21­
day)
12%
TRR
(
28­
day)
Tomato
(
Foliage)
2­
7%
TRR
(
all
PHIs)
N­[
5­
amino­
2­
methyl­
6­
(
2,3,4,5,6­
pentahydroxycyclohexyloxy
tetrahydropyran
3­
yl]­
oxalamic
acid
TABLE
3.4
Tabular
Summary
of
Kasugamycin
Metabolites
and
Degradates.

Common
Name
[
Code]
%
Total
Radioactive
Residues
Chemical
Name
Chemical
Structure
Page
11
of
48
O
O
OH
OH
OH
OH
O
H
C
H
3
N
H
2
NH
2
.
2
HCl
O
O
OH
OH
OH
OH
O
H
C
H
3
NH
NH
O
H
O
NHCOCH
3
Kasuganobiosamine
°
2HCl
[
KB­
2]

Tomato
(
Foliage)
1%
TRR
(
1­
to
28­
day)
6­(
3,5­
diamino­
6­
methyltetrahydro
pyran­
2­
yloxy)­
cyclohexane­
1,2,3,4,5­
pentol
hydrochlorate
2­
N­
acetyl­
kasugamycin
[
KN­
2]

Tomato
(
Foliage)
1­
2%
TRR
(
14­,
21­,
28­
day)
[
5­
acetylamino­
2­
methyl­
6­
(
2,3,4,5,6­
pentahydroxycyclohexyloxy
tetrahydropyran
3­
ylamino]­
iminoacetic
acid
Tomato
metabolism:
MRID
#
45910006.
Single,
foliar
broadcast
application
of
[
14C]­
kasugamycin
to
tomato
plants
at
0.17
lb
ai/
A
(
roughly
3X
the
proposed
maximum
seasonal
rate);
fruit
and
foliage
harvested
at
PHIs
of
2
hours,
1,
7,
14,
21,
and
28
days.
Rat
metabolism:
MRID
#
46030306.
Oral
gavage
dosing
of
[
14C]­
kasugamycin
at
a
single
low
dose
(
100
mg/
kg
bw);
a
single
radiolabeled
high
dose
(
1000
mg/
kg
bw);
repeated
unlabeled
low
doses
(
100
mg/
kg
bw)
for
14
days
in
the
diet,
plus
100
mg/
kg
bw
radiolabeled
single
dose;
or
repeated
unlabeled
high
doses
(
1000
mg/
kg
bw)
for
14
days
in
the
diet,
plus
1000
mg/
kg
bw
radiolabeled
single
dose.

3.5
Toxicity
Profile
of
Major
Metabolites
and
Degradates
No
toxicity
data
were
provided
for
the
metabolites
of
kasugamycin
that
were
observed
in
tomatoes.

3.6
Summary
of
Residues
for
Tolerance
Expression
and
Risk
Assessment
TABLE
3.6
Summary
of
Metabolites
and
Degradates
to
be
included
in
the
Risk
Assessment
and
Tolerance
Expression.

Crop
[
Matrix]
Residue
in
Risk
Assessment
Residue
in
Tolerance
Expression
Fruiting
Vegetables
(
Crop
Group
8)
[
Fruit,
Puree,
Paste]
Parent
compound
only,
kasugamycin
per
se.
Parent
compound
only,
kasugamycin
per
se.

The
only
metabolite
which
would
possibly
be
considered
for
inclusion
in
the
ROC
(
in
addition
to
the
parent)
was
kasugamycinic
acid,
which
was
present
at
less
than
10%
in
all
foliage
Page
12
of
48
samples,
and
only
occurred
at
concentrations
of
10%
or
more
in
fruit
samples
from
the
21­
and
28­
day
PHIs.
Because
the
petitioner
is
proposing
a
PHI
of
1
day
on
imported
fruiting
vegetables,
kasugamycinic
acid
is
unlikely
to
occur
as
other
than
a
minor
constituent
in
imported
fruiting
vegetable
commodities,
and
was
therefore
not
included
in
the
ROC
for
either
tolerance
expression
or
risk
assessment
purposes.
Also,
as
residues
of
kasugamycin
in
all
samples
from
the
crop
field
trials
(
at
1X)
were
either
non­
detectable
or
less
than
the
LOQ,
the
increase
in
risk
from
inclusion
of
this
relatively
minor
metabolite
was
expected
to
be
negligible.

4.0
Hazard
Characterization/
Assessment
4.1
Hazard
Characterization
Kasugamycin
is
a
fungicide
that
belongs
to
the
aminoglycoside
class
of
compounds,
a
group
of
antibiotics
that
inhibit
protein
synthesis.
Unlike
other
members
of
this
class,
kasugamycin
exhibits
activity
only
towards
phytopathogens
and
is
ineffective
against
human
and
animal
pathogens.
As
such,
it
is
not
used
as
a
human
or
veterinary
antibiotic.
In
general,
the
aminoglycosides
are
linked
to
nephrotoxicity.
However,
the
only
evidence
of
nephrotoxicity
seen
in
the
kasugamycin
toxicology
database
was
an
increase
in
kidney
lesions
seen
in
females
in
a
subchronic
mouse
study.
An
acute
oral
study
was
conducted
using
technical
grade
kasugamycin
hydrochloride
hydrate.
Kasumin
Liquid
(
2.02%
ai),
an
end­
use
product
(
EUP),
was
tested
for
acute
oral
toxicity,
dermal
toxicity,
eye
irritation,
dermal
irritation,
and
skin
sensitization.
A
second
EUP,
Kasumin
2
L
(
2.15%
ai),
was
tested
for
acute
inhalation
toxicity.
The
technical
material
has
low
acute
toxicity
in
rats
(
Category
IV
for
acute
oral
exposure).
The
EUPs
also
have
low
acute
toxicity
(
Category
III
for
acute
dermal
exposure
and
Category
IV
for
acute
oral
and
inhalation
exposures).
The
EUP
tested
is
not
a
skin
or
eye
irritant
(
Category
IV),
nor
is
it
a
dermal
sensitizer.
Decreased
body
weights
and
body
weight
gains
were
observed
following
exposure
to
kasugamycin
hydrochloride
hydrate
in
multiple
studies
in
the
rat,
mouse,
dog,
and
rabbit.
In
the
rat,
body
weight
changes
were
reported
in
the
subchronic,
chronic,
developmental,
and
twogeneration
reproduction
studies;
effects
were
seen
at
lower
doses
in
the
longer
term
rat
studies.
In
the
mouse
and
the
dog,
decreases
in
body
weight
and
body
weight
gain
were
seen
in
the
subchronic
studies
at
doses
higher
than
the
LOAEL;
no
treatment­
related
effects,
including
body
weight
changes,
were
noted
in
either
the
chronic
mouse
or
dog
studies.
The
most
sensitive
species
in
terms
of
body
weight
changes
was
the
rabbit.
In
the
developmental
rabbit
study,
the
dose
producing
decreased
body
weights
was
at
least
an
order
of
magnitude
lower
than
the
doses
resulting
in
these
changes
in
the
studies
utilizing
other
species.
In
the
subchronic
dog
study,
the
decreases
in
body
weights
and
body
weight
gains
were
most
likely
associated
with
treatment­
related
effects
on
the
tongue
and
mouth.
These
changes
were
seen
in
both
sexes,
were
dose­
dependent,
and
included
excessive
salivation,
swollen
mouth
with
thickened
skin,
tongue
erosions
and
ulcers,
and
histopathological
lesions
of
the
tongue.
These
effects
may
have
contributed
to
the
decrease
in
food
consumption
observed
in
this
study,
which
likely
led
to
the
decrease
in
body
weights
and
body
weight
gains.
None
of
the
mouth
and
tongue
changes
listed
above
were
seen
in
the
chronic
dog
study,
although
the
highest
dose
tested
Page
13
of
48
in
the
chronic
study
did
cause
those
effects
in
the
subchronic
dog
study.
In
the
developmental
rabbit
study,
the
decreases
in
body
weights
and
body
weight
gains
were
associated
with
decreased
food
consumption.
In
the
main
rabbit
study,
abortions
occurred
in
animals
following
severe
changes
in
body
weights
and
food
consumption.
At
the
doses
tested
in
this
study,
the
abortions
were
not
considered
treatment­
related
because
the
percentage
of
animals
that
aborted
fell
within
the
range
of
the
historical
control
data.
In
two
preliminary
rabbit
studies,
in
which
higher
doses
were
tested,
there
was
a
dose­
dependent
increase
in
mortality
and
abortions
that
accompanied
the
effects
on
body
weights.
No
increased
quantitative
or
qualitative
susceptibility
was
observed
in
the
developmental
rat
or
rabbit
studies.
In
the
developmental
rat
study,
no
developmental
toxicity
was
observed
at
the
highest
dose
tested,
although
this
dose
level
induced
toxicity
in
the
dams
including
loose
stool,
distention
of
the
large
intestine,
and
decreased
maternal
body
weights,
body
weight
gains,
and
food
consumption.
In
the
main
developmental
rabbit
study,
neither
offspring
nor
maternal
toxicity
was
observed
up
to
the
highest
dose
tested.
Although
abortions
were
seen
in
the
main
study,
the
percentage
of
animals
that
aborted
fell
within
the
range
of
historical
controls.
Higher
dose
levels
were
tested
in
the
preliminary
developmental
rabbit
studies.
In
these
studies,
dose­
dependent
increases
in
late­
term
abortions
(
and
maternal
mortality)
were
observed
in
animals
following
severe
changes
in
maternal
body
weights
associated
with
decreased
food
consumption.
Maternal
toxicity,
indicated
by
the
decreases
in
food
consumption
and
body
weights,
was
seen
following
multiple
doses
of
kasugamycin
hydrochloride
hydrate.
In
the
first
preliminary
rabbit
developmental
toxicity
study,
in
which
all
treated
animals
were
sacrificed
in
extremis,
all
of
the
animals
survived
for
at
least
8
days
of
dosing
up
to
the
limit
dose
(
1000
mg/
kg/
day);
there
were
no
abortions
in
this
study.
Because
the
abortions
only
occurred
after
maternal
toxicity
developed
following
multiple
doses
of
kasugamycin
hydrochloride
hydrate,
the
abortions
seen
in
the
rabbit
developmental
toxicity
study
are
not
attributable
to
a
single
oral
dose.
Increased
qualitative
or
quantitative
susceptibility
was
also
not
observed
in
the
twogeneration
reproduction
study
in
rats.
In
this
study,
decreased
parental­
generation
male
body
weights
and
body
weight
gains
were
noted
at
a
lower
dose
than
the
lowest
dose
at
which
effects
on
reproduction
were
observed.
Reproductive
toxicity
was
only
observed
in
the
F1
generation
and
included
decreased
fertility
and
fecundity
in
males
and
females,
as
well
as
an
increased
precoital
interval
during
the
mating
period
for
the
F2
litter.
No
offspring
toxicity
was
found
at
any
of
the
doses
tested
in
the
two­
generation
reproduction
study;
however,
offspring
sexual
maturation,
organ
weights,
and
histopathology
were
not
examined.
Testicular
effects
were
noted
in
multiple
studies.
In
the
combined
chronic/
carcinogenicity
study
in
rats,
the
basis
for
the
LOAEL
was
an
increase
in
the
incidence
and
severity
of
testicular
tubular
atrophy
noted
during
the
histopathological
examinations
at
the
end
of
the
dosing
period
(
ie,
week
104),
as
well
as
at
two
interim
time
points
(
ie,
weeks
26
and
52).
Also
in
the
chronic
rat
study,
the
incidences
of
testicular
atrophy
and
softening
observed
grossly
in
males
at
the
LOAEL
were
increased,
compared
to
controls.
As
mentioned
previously,
reproductive
toxicity
(
ie,
decreased
fertility,
decreased
fecundity,
and
an
increase
in
the
pre­
coital
interval
during
the
mating
period
for
the
F2
litter)
was
seen
in
rats
in
the
two­
generation
reproduction
study.
In
the
reproduction
study,
small,
fluid­
filled
testes
were
observed
in
the
F1
male
rats.
At
the
microscopic
level,
these
animals
had
severe
testicular
atrophy
and
degeneration,
with
a
complete
loss
of
the
germinal
epithelium.
In
the
subchronic
mouse
study,
an
increased
number
of
high­
dose
males
had
testicular
tubular
dilatation
and
degeneration
compared
to
controls.
No
treatmentrelated
effects,
including
testicular
changes,
were
observed
in
the
chronic
mouse
study;
however,
Page
14
of
48
the
highest
dose
tested
in
this
study
was
half
of
the
dose
selected
as
the
LOAEL
in
the
subchronic
mouse
study.
In
the
subchronic
dog
study,
1/
4
of
the
high
dose
dogs
had
hypospermia
compared
to
0/
4
of
the
other
dose
groups.
In
the
chronic
dog
study,
2/
4
of
the
high
dose
males
had
testicular
degeneration
and
3/
4
had
chronic
inflammation
of
the
testes;
only
1/
4
of
the
controls
had
degeneration
and
0/
4
had
inflammation.
No
mutagenic
potential
was
noted
in
the
following
acceptable/
guideline
mutagenicity
studies:
(
1)
the
in
vitro
mammalian
cell
forward
gene
mutation
test,
(
2)
the
mammalian
erythrocyte
micronucleus
test,
and
(
3)
the
unscheduled
DNA
synthesis
in
mammalian
cells
in
culture.
Additionally,
no
mutagenic
potential
was
observed
in
an
acceptable/
non­
guideline
host­
mediated
gene
mutation
assay.
Two
of
the
submitted
mutagenicity
studies
were
classified
unacceptable.
The
bacterial
gene
mutation
assay
(
MRID
#
45910028)
was
classified
as
such
because
kasugamycin
hydrochloride
hydrate
was
not
tested
up
to
the
limit
dose
(
5000

g/
plate);
however,
no
mutagenic
potential
was
noted
up
to
the
highest
dose
that
was
tested
in
the
study
(
500

g/
plate).
The
structural
chromosomal
aberration
study
(
MRID
#
45910025)
was
classified
unacceptable
because
there
were
only
10
hours
(
ie,
less
than
1
cell
cycle)
between
the
time
from
treatment
to
cell
harvest,
rather
than
the
recommended
1.5
cell
cycles
(
ie,
16
to
21
hours);
however,
there
was
no
evidence
that
kasugamycin
hydrochloride
hydrate
induced
a
clastogenic
effect.
Although
these
two
unacceptable
mutagenicity
studies
create
a
data
gap,
in
the
absence
of
a
carcinogenic
effect
in
the
rat
and
mouse
studies
and
of
a
mutagenic
effect
in
the
available
studies,
the
concern
for
mutagenesis
is
reduced.
Therefore,
on
4/
13/
2005,
the
Risk
Assessment
Review
Committee
(
RARC)
recommended
that
the
requirement
for
these
mutagenicity
studies
be
waived
for
the
establishment
of
tolerances
for
kasugamycin
on
import
commodities
only.
At
this
time,
an
acceptable
bacterial
gene
mutation
assay
and
an
acceptable
in
vitro
structural
chromosomal
aberration
study
will
not
further
inform
the
risk
assessment.
No
oncogenic
potential
was
noted
in
the
mouse
oncogenicity
study
or
in
the
rat
combined
chronic/
carcinogenicity
study;
additionally,
no
mutagenic
potential
was
noted
in
any
of
the
available
mutagenicity
studies.
Based
on
the
overall
weight
of
the
evidence,
kasugamycin
is
classified
as
"
not
likely
to
be
carcinogenic
to
humans."
Other
effects
seen
in
the
toxicology
database
for
kasugamycin
include
anal
lesions
and
perianal/
perigenital
staining
in
the
subchronic
mouse
study.
Similarly,
red
and
swollen
skin
around
the
anal
opening
and
inflammation
and
ulceration
of
the
rectum
were
noted
in
rats
of
both
sexes
and
both
generations
in
the
two­
generation
reproduction
study.
These
effects
may
be
related
to
the
acidity
of
the
active
ingredient,
which
is
primarily
eliminated
in
the
feces.
The
parent
compound
does
not
undergo
significant
metabolism;
less
than
1%
of
the
administered
dose
was
identified
in
the
urine,
liver,
kidneys,
and
plasma
as
the
metabolite
kasuganobiosamine.

TABLE
4.1a
Acute
Toxicity
Profile
for
Kasugamycin.

Test
Material*
[%
ai]
Guideline
Number
Study
Type
MRID
Number
Results
Toxicity
Category
Technical
Product
[
71]
870.1100
Acute
oral
­
rat
45910012
LD50
(

+

)
>
5000
mg/
kg
IV
EUP
[
2.0]
870.1100
Acute
oral
­
rat
45910014
LD50
(

+

)
>
5000
mg/
kg
IV
EUP
[
2.0]
870.1100
Acute
oral
­
mouse
45910013
LD50
(

+

)
>
5000
mg/
kg
IV
TABLE
4.1a
Acute
Toxicity
Profile
for
Kasugamycin.

Test
Material*
[%
ai]
Guideline
Number
Study
Type
MRID
Number
Results
Toxicity
Category
Page
15
of
48
EUP
[
2.0]
870.1200
Acute
dermal
­
rat
46030301
LD50
(

+

)
>
2000
mg/
kg
III
EUP
[
2.2]
870.1300
Acute
inhalation
­
rat
45910018
LC50
(

+

)
>
4.892
mg/
L
IV
EUP
[
2.0]
870.2400
Acute
eye
irritation
­
rabbit
45910015
Mild
eye
irritant
(
iritis
at
1
hour,
resolving
by
24
hours;
conjunctivitis
at
1
hour,
resolving
by
24
hours).
IV
EUP
[
2.0]
870.2500
Acute
dermal
irritation
­
rabbit
45910017
Not
irritating
to
the
skin.
IV
EUP
[
2.0]
870.2600
Skin
sensitization
­
guinea
pig
45910016
Not
a
sensitizer
under
the
conditions
of
this
study.
Not
applicable
*
EUPs
are
formulated
using
kasugamycin
hydrochloride
hydrate;
technical
product
(
Lot
#
KP­
913)
is
81%
kasugamycin
hydrochloride
hydrate.
Bracketed
values
are
%
ai
as
kasugamycin
free
base.

TABLE
4.1b
Subchronic,
Chronic,
and
Other
Toxicity
Profile
for
Kasugamycin.

Guideline
Number
Study
Type/
Classification
MRID
Number
Doses
Results
870.3100
90­
Day
oral
toxicity
rodents
­
rat
Acceptable/
guideline
45910020
0,
300,
1000,
3000,
6000
ppm
M:
0,
17.5,
58.2,
176.7,
354.8
mg/
kg/
day
F:
0,
20.3,
69.2,
201.0,
395.5
mg/
kg/
day
NOAEL
=
176.7/
201.0
mg/
kg/
day
(
M/
F)
LOAEL
=
354.8/
395.5
mg/
kg/
day
(
M/
F)
based
on
decreased
body
weights
and
body
weight
gains.

870.3100
90­
Day
oral
toxicity
rodents
­
mouse
Acceptable/
guideline
45910019
0,
300,
1000,
3000,
10000
ppm
M:
0,
41.2,
135.4,
408.5,
1559
mg/
kg/
day
F:
0,
58.0,
170.9,
565.6,
1834
mg/
kg/
day
NOAEL
=
135.4/
170.9
mg/
kg/
day
(
M/
F)
LOAEL
=
408.5/
565.6
mg/
kg/
day
(
M/
F)
based
on
increased
mortality
and
anal
lesions
(
M&
F),
and
kidney
lesions
(
F).

At
1559/
1834
mg/
kg/
day
(
M/
F),
decreased
body
weights
and
body
weight
gains
(
M&
F),
testicular
tubular
dilatation
and
degeneration,
perianal/
perigenital
staining
(
F),
and
extramedullary
hematopoiesis
of
the
spleen
(
M)
were
seen.
TABLE
4.1b
Subchronic,
Chronic,
and
Other
Toxicity
Profile
for
Kasugamycin.

Guideline
Number
Study
Type/
Classification
MRID
Number
Doses
Results
Page
16
of
48
870.3150
90­
Day
oral
toxicity
in
nonrodents
­
dog
Acceptable/
guideline
46030302
0,
300,
3000,
6000/
0/
4500*
ppm
M:
0,
10.6,
106.0,
182
mg/
kg/
day
F:
0,
11.4,
107.9,
179
mg/
kg/
day
*
The
high­
dose
group
was
exposed
to
6000
ppm
on
weeks
1­
5,
control
diet
on
weeks
6­
8,
and
4500
ppm
on
weeks
8­
13.
NOAEL
=
10.6/
11.4
mg/
kg/
day
(
M/
F)
LOAEL
=
106.0/
107.9
mg/
kg/
day
(
M/
F)
based
on
tongue
lesions,
few
feces,
swollen
mouth,
excessive
salivation,
and
thickened
skin
at
the
commissure
of
the
mouth.

At
182/
170
mg/
kg/
day
(
M/
F),
decreased
body
weights,
body
weight
gains,
and
food
consumption
were
seen.

870.3200
21/
28­
Day
dermal
toxicity
Not
applicable
Not
performed
870.3250
90­
Day
dermal
toxicity
Not
applicable
Not
performed
870.3465
90­
Day
inhalation
toxicity
Not
applicable
Not
performed
870.3700
Pre­
natal
developmental
in
rodents
­
rat
Acceptable/
guideline
45910022
0,
40,
200,
1000
mg/
kg/
day
Maternal
NOAEL
=
200
mg/
kg/
day
LOAEL
=
1000
mg/
kg/
day
based
on
decreased
body
weights,
body
weight
gains,
and
food
consumption;
increased
incidence
of
loose
stool;
and
distention
of
the
large
intestine
with
stool
in
the
cecum.

Developmental
NOAEL
=
1000
mg/
kg/
day
LOAEL
=
>
1000
mg/
kg/
day
870.3700
Pre­
natal
developmental
in
nonrodents
­
rabbit
Acceptable/
guideline
46030303
0,
1,
3,
10
mg/
kg/
day
Maternal
NOAEL
=
10
mg/
kg/
day
LOAEL
=
>
10
mg/
kg/
day
Note:
Abortions
and
decreased
maternal
body
weights,
body
weight
gains,
and
food
consumption
were
seen
at
30
mg/
kg/
day
in
a
range­
finding
study.

Developmental
NOAEL
=
10
mg/
kg/
day
LOAEL
=
>
10
mg/
kg/
day
TABLE
4.1b
Subchronic,
Chronic,
and
Other
Toxicity
Profile
for
Kasugamycin.

Guideline
Number
Study
Type/
Classification
MRID
Number
Doses
Results
Page
17
of
48
870.3800
Reproduction
and
fertility
effects
­
rat
Acceptable/
guideline
45910023
0,
200,
1000,
6000
ppm
M:
0,
13.7,
70.3,
425.3
mg/
kg/
day
F:
0,
16.2,
82.9,
503.4
mg/
kg/
day
Parental/
Systemic
NOAEL
=
13.7/
16.2
mg/
kg/
day
(
M/
F)
LOAEL
=
70.3/
82.9
mg/
kg/
day
(
M/
F)
based
on
decreased
body
weights
and
body
weight
gains.

At
425.3/
503.4
mg/
kg/
day
(
M/
F),
red
and
swollen
skin
around
the
anal
opening
(
M&
F)
and
testicular
atrophy/
degeneration
in
F1
males
were
seen.

Reproductive
NOAEL
=
70.3/
82.9
mg/
kg/
day
(
M/
F)
LOAEL
=
425.3/
503.4
mg/
kg/
day
(
M/
F)
based
on
decreased
fertility
and
fecundity
in
the
F1
parents
for
both
litters
and
increased
pre­
coital
interval
during
the
mating
period
for
the
F2
litter.

Offspring
NOAEL
=
425.3/
503.4
mg/
kg/
day
(
M/
F)
LOAEL
=
>
425.3/
503.4
mg/
kg/
day
(
M/
F)

870.4100
Chronic
toxicity
­
rodents
See
870.4300.
This
study
includes
requirements
of
both
870.4100
and
870.4200.

870.4100
Chronic
toxicity
­
dog
Acceptable/
guideline
46185901
0,
300,
1000,
3000
ppm
M:
0,
10.5,
30.5,
99.6
mg/
kg/
day
F:
0,
9.4,
33.4,
103.6
mg/
kg/
day
NOAEL
=
99.6/
103.6
mg/
kg/
day
(
M/
F)
LOAEL
=
>
99.6/
103.6
mg/
kg/
day
(
M/
F)

870.4200
Carcinogenicity
­
rat
See
870.4300.
This
study
includes
requirements
of
both
870.4100
and
870.4200.

870.4200
Carcinogenicity
­
mouse
Acceptable/
guideline
46030304
0,
50,
300,
1500
ppm
M:
0,
5.93,
34.94,
186.3
mg/
kg/
day
F:
0,
7.25,
42.29,
215.2
mg/
kg/
day
NOAEL
=
186.3/
215.2
mg/
kg/
day
(
M/
F)
LOAEL
=
>
186.3/
215.2
mg/
kg/
day
(
M/
F)

No
evidence
of
carcinogenicity
TABLE
4.1b
Subchronic,
Chronic,
and
Other
Toxicity
Profile
for
Kasugamycin.

Guideline
Number
Study
Type/
Classification
MRID
Number
Doses
Results
Page
18
of
48
870.4300
Combined
chronic
toxicity/
carcinogenicity
­
rat
Acceptable/
guideline
45910024
0,
30,
300,
3000
ppm
M:
0,
1.1,
11.3,
116
mg/
kg/
day
F:
0,
1.4,
13.4,
140
mg/
kg/
day
NOAEL
=
11.3/
140
mg/
kg/
day
(
M/
F)
LOAEL
=
116/>
140
mg/
kg/
day
(
M/
F)
based
on
increased
testicular
softening
and
atrophy
in
males.

No
evidence
of
carcinogenicity
870.5100
Gene
mutation
­
bacterial
reverse
mutation
assay
Unacceptable/
upgradable
45910028
0,
5,
10,
50,
100,
500
ug/
plate
for
Salmonella
typhimurium
strain
G46
(
his­)

0,
5,
10,
50,
100,
200
ug/
plate
for
all
other
strains
tested
No
mutagenic
activity
in
bacteria
(
Salmonella
typhimurium
and
Escherichia
coli)
under
conditions
of
this
assay.

Not
tested
up
to
the
limit
dose,
no
indication
of
cytotoxicity,
and
no
defined
limit
of
solubility.

870.5300
Cytogenetics
­
in
vitro
mammalian
cell
gene
mutation
test
(
CHO
Cells)

Acceptable/
guideline
45910026
0,
0.5,
1,
2,
4,
6,
8,
10
mg/
ml
No
increase
in
mutant
frequency
at
the
HGPRT
locus,
in
the
presence
or
absence
of
S9
activation.

870.5375
Cytogenetics
­
in
vitro
mammalian
cell
chromosome
aberration
test
Unacceptable/
not
upgradable
45910025
0,
1,
2,
3,
4,
5
mg/
ml
No
increase
in
mutant
frequency,
in
the
presence
or
absence
of
S9
activation.

The
time
from
treatment
to
cell
harvest
was
insufficient.

870.5395
Cytogenetics
­
mammalian
erythrocyte
micronucleus
test
(
mice)

Acceptable/
guideline
46030305
0,
200,
1000,
5000
mg/
kg
No
evidence
of
induced
chromosomal
damage
or
other
damage
leading
to
micronucleus
formation.

870.5550
Other
effects
­
unscheduled
DNA
synthesis
in
mammalian
cells
in
culture
(
rats)

Acceptable/
guideline
45910027
First
assay:
0­
2.5
mg/
ml
Second
assay:
0­
10
mg/
ml
Third
assay:
0­
10
mg/
ml
No
evidence
that
unscheduled
DNA
synthesis
was
induced.
TABLE
4.1b
Subchronic,
Chronic,
and
Other
Toxicity
Profile
for
Kasugamycin.

Guideline
Number
Study
Type/
Classification
MRID
Number
Doses
Results
Page
19
of
48
870.7485
Metabolism
and
pharmacokinetics
­
rat
Acceptable/
guideline
46030306
(
1)
100
mg/
kg
radiolabeled,
single
dose
by
oral
gavage.

(
2)
100
mg/
kg
unlabeled,
14
days
in
the
diet,
PLUS
100
mg/
kg
radiolabeled,
single
dose
by
oral
gavage.

(
3)
1000
mg/
kg
radiolabeled,
single
dose
by
oral
gavage.

(
4)
1000
mg/
kg
unlabeled,
14
days
in
the
diet,
PLUS
1000
mg/
kg
radiolabeled,
single
dose
by
oral
gavage.
The
mean
radioactivity
recovery
168
hours
after
exposure
ranged
between
90.6­
96.7%,
with
the
majority
of
the
dose
recovered
within
48
hours
in
the
feces
(
81.9­
93.9%)
and
urine
(
1.26­
3.07%).
The
maximum
concentration
found
in
the
plasma
of
both
males
and
females
occurred
approximately
one
hour
after
the
administration
of
a
single
low
or
high
dose.
Between
one
and
six
hours
after
a
single
low
or
high
dose,
more
kasugamycin
accumulated
in
the
kidneys,
urinary
bladder,
and
lymph
nodes
than
in
the
blood,
but
after
168
hours,
little
or
no
kasugamycin
was
found
in
these
tissues.
The
absorption
and
metabolism
of
kasugamycin
in
rats
was
limited
(<
5%
dose)
and
was
not
affected
by
sex,
dose
level,
or
duration
of
dosing.
Parent
compound
was
the
major
component
identified
in
the
urine,
feces,
liver,
kidney,
and
plasma.
Minor
amounts
(<
1%
dose)
of
the
metabolite
kasuganobiosamine
were
identified
in
urine,
liver,
kidney,
and
plasma,
but
none
was
detected
in
the
feces.
Elimination
occurred
primarily
in
the
feces
(
87.7­
94.5%);
however,
kasugamycin
was
not
excreted
in
the
bile
(
enterohepatic
circulation
did
not
occur).

4.2
FQPA
Hazard
Considerations
4.2.1
Adequacy
of
the
Toxicity
Data
Base
The
toxicology
database
for
kasugamycin
is
considered
adequate
in
terms
of
endpoint
and
dose­
response
information
to
characterize
any
potential
pre­
and/
or
post­
natal
risk
for
infants
and
children.
Acceptable
rat
and
rabbit
developmental
toxicity
studies
and
an
acceptable
twogeneration
reproduction
study
in
the
rat
were
submitted.

4.2.2
Evidence
of
Neurotoxicity
There
is
no
evidence
that
exposure
to
kasugamycin
results
in
neurotoxicity.
No
clinical
Page
20
of
48
signs
of
neurotoxicity,
changes
in
brain
weights,
changes
in
brain
histopathology,
or
any
other
neurotoxic
effects
were
observed
in
any
of
the
submitted
studies,
including
the
subchronic
feeding
studies,
the
chronic
feeding
studies,
the
developmental
toxicity
studies,
or
the
two­
generation
reproduction
study.

4.2.3
Developmental
Toxicity
Studies
No
increased
quantitative
or
qualitative
susceptibility
was
observed
in
the
developmental
rat
or
rabbit
studies.
In
the
developmental
rat
study,
no
developmental
toxicity
was
observed
at
the
highest
dose
tested,
although
this
dose
level
induced
toxicity
in
the
dams
including
loose
stool,
distention
of
the
large
intestine,
and
decreased
maternal
body
weights,
body
weight
gains,
and
food
consumption.
In
the
main
developmental
rabbit
study,
neither
offspring
nor
maternal
toxicity
was
observed
up
to
the
highest
dose
tested.
Although
abortions
were
seen
in
the
main
study,
the
percentage
of
animals
that
aborted
fell
within
the
range
of
historical
controls.
Higher
dose
levels
were
tested
in
the
preliminary
developmental
rabbit
studies.
In
these
studies,
dose­
dependent
increases
in
late­
term
abortions
(
and
maternal
mortality)
were
observed
in
animals
following
severe
changes
in
maternal
body
weights
associated
with
decreased
food
consumption.

4.2.3.1
Developmental
Toxicity
Study
in
Rats
In
a
developmental
toxicity
study
(
MRIDs
#
45910022
and
45910021),
kasugamycin
hydrochloride
hydrate
(
64.5%
ai,
Lot
#
KP­
834)
in
purified
water
was
administered
daily
by
oral
gavage
to
24
female
Crj:
CD(
SD)
rats/
dose
in
a
dose
volume
of
10
mL/
kg
bw
at
dose
levels
of
0,
40,
200,
or
1000
mg/
kg
bw/
day
on
gestation
days
(
GD)
6
through
15.
All
dams
were
killed
on
GD
20;
their
fetuses
were
removed
by
cesarean
section
and
examined.
It
was
assumed,
but
not
explicitly
stated,
that
doses
were
adjusted
for
purity
of
the
test
compound.
All
animals
survived
to
scheduled
termination.
For
maternal
toxicity,
there
were
treatment­
related
effects
on
clinical
signs,
body
weight,
and
food
consumption
at
1000
mg/
kg/
day.
An
increased
incidence
(
p

0.01)
of
loose
stool
was
sporadically
observed
during
GD
8
to
15
in
7
high­
dose
dams
(
9
observations),
compared
to
none
of
the
controls.
An
increased
incidence
(
p

0.05)
of
distention
of
the
large
intestine
with
stool
in
the
cecum
was
observed
in
5
of
24
high­
dose
dams,
compared
with
none
of
the
controls.
Body
weights
were
decreased
in
the
high­
dose
group
(­
3
to
­
7%;
p

0.05)
during
GD
8
to
20,
which
resulted
in
decreased
body
weight
gains
during
the
dosing
period
(­
54%
on
GD
6
to
15;
calculated
by
reviewers)
and
during
the
overall
study
(­
17%
on
GD
0
to
20;
p

0.01).
The
gravid
uterine
weights
of
the
high­
dose
dams
were
non­
significantly
decreased
by
7%.
When
adjusted
for
gravid
uterine
weight,
the
body
weights
of
these
females
were
decreased
on
GD
20
(­
5%;
p

0.05).
Food
consumption
was
decreased
during
the
dosing
period
(­
24%
on
GD
6
to
15;
p

0.05).
The
maternal
LOAEL
is
1000
mg/
kg/
day
(
limit
dose),
based
on
decreased
body
weights,
body
weight
gains,
and
food
consumption,
and
increased
incidences
of
loose
stool
and
of
distention
of
the
large
intestine
with
stool
in
the
cecum.
The
maternal
NOAEL
is
200
mg/
kg/
day.
For
developmental
toxicity,
there
were
no
abortions
or
premature
deliveries.
There
were
no
treatment­
related
effects
on
the
number
of
litters,
number
of
fetuses
(
live
or
dead),
number
of
resorptions
(
early,
late,
or
complete
litter),
post­
implantation
loss,
or
fetal
sex
ratio.
Although
placental
weights
were
decreased
at
1000
mg/
kg/
day
(­
12%;
p

0.01),
there
were
no
treatmentrelated
effects
on
fetal
body
weights,
so
this
finding
is
considered
equivocal.
There
were
no
Page
21
of
48
treatment­
related
external,
visceral,
or
skeletal
malformations
or
variations.
The
developmental
LOAEL
was
not
observed.
The
developmental
NOAEL
is
1000
mg/
kg/
day
(
limit
dose).
This
study
is
classified
acceptable/
guideline
and
satisfies
the
guideline
requirements
for
a
developmental
toxicity
study
(
OPPTS
870.3700a;
OECD
414)
in
the
rat.

4.2.3.2
Developmental
Toxicity
Study
in
Rabbits
In
a
developmental
toxicity
study
(
MRID
#
46030303),
kasugamycin
hydrochloride
hydrate
(
62.7%
ai,
Lot
#
KP­
570)
in
distilled
water
was
administered
daily
by
oral
gavage
to
15
female
New
Zealand
White
rabbits/
dose
in
a
dose
volume
of
10
mL/
kg
at
dose
levels
of
0,
1,
3,
or
10
mg/
kg
bw/
day
on
GD
6
through
19.
All
surviving
does
were
killed
on
GD
29;
their
fetuses
were
removed
by
cesarean
section
and
examined.
In
the
main
study,
there
were
no
treatment­
related
effects
on
maternal
mortality,
clinical
signs,
body
weight,
food
consumption,
or
gross
pathology.
Although
effects
were
not
seen
at
the
highest
dose
tested,
a
new
developmental
toxicity
study
in
the
rabbit
is
not
required
because
treatment­
related
effects
were
seen
at
dose
levels
of
30
mg/
kg/
day
and
higher
in
the
preliminary
studies
(
MRID
#
46428701).
The
maternal
LOAEL
was
not
observed.
The
maternal
NOAEL
is
10
mg/
kg/
day.
There
were
no
treatment­
related
effects
on
placental
weight,
fetal
weight,
sex
ratio,
postimplantation
loss,
or
the
numbers
of
litters,
fetuses
(
live
or
dead),
or
resorptions
(
early,
late,
or
complete
litter).
There
were
no
treatment­
related
external,
visceral,
or
skeletal
malformations,
variations,
or
retardations.
The
developmental
LOAEL
was
not
observed.
The
developmental
NOAEL
is
10
mg/
kg/
day.
This
study
is
classified
acceptable/
guideline
and
satisfies
the
guideline
requirements
for
a
developmental
toxicity
study
(
OPPTS
870.3700b;
OECD
414)
in
the
rabbit.

4.2.4
Reproductive
Toxicity
Study
In
a
two­
generation
reproduction
toxicity
study
(
MRID
#
45910023),
kasugamycin
hydrochloride
hydrate
(
81%
ai;
Lot
#
KP­
913)
was
administered
continuously
in
the
diet
to
Crl:
CD
®
BR
VAF/
Plus
®
rats
(
25
rats/
sex/
dose)
at
dose
levels
of
0,
200,
1000,
or
6000
ppm
(
equivalent
to
0/
0,
13.7/
16.2,
70.3/
82.9,
425.3/
503.4
mg/
kg
bw/
day
[
M/
F]).
The
P
and
F
1
parents
were
dosed
for
10
weeks
before
they
were
mated
to
produce
the
F
1
and
F
2a
litters,
respectively.
The
F
1
pups
were
weaned
on
post­
natal
day
(
PND)
21,
and
25
pups/
sex/
group
(
1
pup/
sex/
litter
as
nearly
as
possible)
were
randomly
selected
as
parents
of
the
F
2
generation.
Because
of
low
fertility
of
the
F
1
parents
during
the
production
of
the
F
2a
litter,
the
F
1
adults
were
paired
again
six
days
following
weaning
of
the
F
2a
litter
to
produce
the
F
2b
litter.
F
1
males
that
did
not
sire
a
litter
in
the
first
(
F
2a)
mating
were
paired
with
proven
females,
and
females
that
failed
to
deliver
an
F
2a
litter
were
paired
with
proven
males.
In
the
P
generation,
1000
ppm
male
body
weights
were
decreased
(­
6
to
­
7%;
p

0.05)
during
Weeks
4
to
9
and
Week
11,
and
6000
ppm
male
body
weights
were
decreased
(­
5
to
­
6%;
p

0.05)
during
Weeks
6
to
9.
P
generation
cumulative
body
weight
gains
were
decreased
in
the
1000
ppm
males
for
Weeks
3
to
9
(
p

0.05)
and
in
the
6000
ppm
males
for
Weeks
3
to
14
(
p

0.05).
In
the
F
1
generation,
cumulative
body
weight
gains
were
decreased
in
the
6000
ppm
males
(
p

0.05)
during
Weeks
1
to
3,
5
to
6,
and
9
to
11
of
the
pre­
mating
period
for
the
F
2b
litter.
Red
and
swollen
skin
around
the
anal
opening
was
observed
exclusively
in
the
6000
ppm
Page
22
of
48
group
in
both
sexes
and
in
both
generations.
This
clinical
sign
was
first
noted
at
Week
10
in
the
P
males
(
16
to
28%)
and
P
females
(
13
to
100%),
and
it
was
first
seen
at
Week
5
in
the
F
1
males
(
8
to
75%)
and
F
1
females
(
12
to
100%).
Gross
examination
of
the
animals
in
this
dose
group
revealed
red
foci/
areas
on
the
rectum
in
both
sexes
and
both
generations
(
80
to
100%
treated
vs
0
controls),
as
well
as
thickened
walls
of
the
rectum
in
the
F
1
males
(
46%
treated
vs
0
controls).
Microscopically,
chronic
active
inflammation
and
ulceration
of
the
rectum
were
observed
in
both
sexes
of
both
generations
(
88
to
100%
treated
vs
0
controls).
Additionally,
in
the
P
males
at
this
dose,
squamous
cell
hyperplasia
was
observed
at
the
ano­
rectal
junction
(
38%
treated
vs
0
controls).
At
6000
ppm,
testes
that
were
small
(
38%)
and
containing
fluid
(
67%)
were
observed
(
compared
to
0%
of
controls)
in
the
F
1
males
at
necropsy.
Microscopically,
atrophy/
degeneration
was
noted
in
the
testes
in
the
P
(
12%
treated
vs
0%
controls)
and
F
1
(
63%
treated
vs
0%
controls)
males.
This
atrophy/
degeneration
was
marked
to
severe
in
the
F
1
generation
and
was
characterized
by
complete
loss
of
the
germinal
epithelium,
atrophy
of
the
seminiferous
tubules,
persistence
of
Sertoli
cells,
and
variable
amounts
of
intertubular
edema.
Additionally
in
the
6000
ppm
F
1
males,
pelvic
dilatation
of
the
kidney
and
chronic
progressive
nephropathy
were
observed
microscopically
(
21%
treated
vs
4%
controls).
The
LOAEL
for
parental
toxicity
is
1000
ppm
(
equivalent
to
70.3/
82.9
mg/
kg/
day
[
M/
F]),
based
on
decreased
body
weights
and
body
weight
gains
in
the
P
generation
males.
The
NOAEL
is
200
ppm
(
equivalent
to
13.7/
16.2
mg/
kg/
day
[
M/
F]).
In
F
1
female
pups,
adjusted
body
weights
were
decreased
(­
6%;
p

0.05)
at
6000
ppm
on
PND
21.
This
decrease
was
minor
and
occurred
only
at
the
end
of
the
lactation
period.
Furthermore,
there
were
no
treatment­
related
effects
on
body
weights
in
the
F
2a
or
F
2b
females
or
in
the
male
pups
of
either
generation
at
this
dose.
Thus,
this
decrease
is
not
considered
adverse.
Pup
weights
at

1000
ppm
in
both
sexes
and
generations
were
comparable
to
controls.
It
should
be
noted
that
several
parameters
were
not
examined
in
the
offspring,
including
sexual
maturation,
developmental
landmarks,
organ
weights,
and
histopathology.
Based
on
the
parameters
investigated,
the
LOAEL
for
offspring
toxicity
was
not
observed.
The
NOAEL
is
6000
ppm
(
equivalent
to
425.3/
503.4
mg/
kg/
day).
In
the
F
1
generation,
male
and
female
fertility
and
fecundity
indices
were
decreased
at
6000
ppm
for
the
F
2a
litter
(
64
to
67%
treated
vs
88
to
96%
controls)
and
the
F
2b
litter
(
36
to
39%
treated
vs
88
to
100%
controls).
Additionally,
in
the
F
1
generation,
the
pre­
coital
interval
during
the
mating
period
to
produce
the
F
2b
litter
was
longer
(
p

0.01)
at
6000
ppm
(
5.73
days)
compared
to
controls
(
2.23
days).
The
LOAEL
for
reproductive
toxicity
was
6000
ppm
(
equivalent
to
425.3/
503.4
mg/
kg/
day
[
M/
F])
based
on
decreased
fertility
and
fecundity
in
the
F1
parents
for
both
litters
and
increased
pre­
coital
interval
during
the
mating
period
for
the
F2
litter.
The
NOAEL
is
1000
ppm
(
equivalent
to
70.3/
82.9
mg/
kg/
day
[
M/
F]).
This
study
is
classified
as
acceptable/
guideline
and
satisfies
the
Guideline
requirements
(
OPPTS
870.3800;
OECD
416)
for
a
two­
generation
reproduction
study
in
the
rat.

4.2.5
Additional
Information
from
Literature
Sources
Studies
available
in
the
open
literature
include
articles
on
the
mechanism
of
resistance
and
toxicity
of
kasugamycin
towards
various
species
of
bacteria.

4.2.6
Pre­
and/
or
Post­
Natal
Toxicity
Page
23
of
48
4.2.6.1
Determination
of
Susceptibility
No
increased
quantitative
or
qualitative
susceptibility
was
observed
in
the
developmental
rat
or
rabbit
studies
or
in
the
two­
generation
reproduction
study.
No
offspring
toxicity
was
observed
at
any
of
the
doses
tested
in
these
three
studies.
Reproductive
toxicity
was
noted
in
the
F1
generation
of
the
two­
generation
reproduction
study.
However,
because
parental
toxicity
(
decreased
body
weights
and
body
weight
gains)
occurred
at
a
lower
dose
than
that
which
resulted
in
effects
on
reproduction,
there
is
no
increased
quantitative
or
qualitative
susceptibility
of
the
offspring.

4.2.6.2
Degree
of
Concern
Analysis
and
Residual
Uncertainties
for
Pre­
and/
or
Post­
Natal
Susceptibility
The
toxicology
database
for
kasugamycin
is
complete
with
respect
to
pre­
and
post­
natal
toxicity
and
shows
no
evidence
of
increased
qualitative
or
quantitative
susceptibility
in
the
offspring.
Therefore,
there
are
no
residual
uncertainties
for
pre­
and/
or
post­
natal
toxicity.

4.3
Recommendation
for
a
Developmental
Neurotoxicity
Study
4.3.1
Evidence
that
Supports
Requiring
a
Developmental
Neurotoxicity
Study
There
is
no
evidence
to
support
the
requirement
for
a
developmental
neurotoxicity
study
on
kasugamycin.

4.3.2
Evidence
that
Supports
not
Requiring
a
Developmental
Neurotoxicity
Study
There
was
no
evidence
of
neurotoxicity
in
any
of
the
studies
available
in
the
toxicology
database,
including
the
subchronic
feeding
studies,
the
chronic
feeding
studies,
the
developmental
toxicity
studies,
and
the
two­
generation
reproduction
study.
Therefore,
acute
and
subchronic
neurotoxicity
studies
are
not
required.
A
developmental
neurotoxicity
study
is
not
required
because:
(
1)
there
are
no
data
gaps
for
the
assessment
of
the
effects
of
kasugamycin
following
in
utero
and/
or
post­
natal
exposure,
(
2)
there
is
no
evidence
of
neurotoxicity
in
the
available
toxicology
studies,
and
(
3)
there
is
no
increased
quantitative
or
qualitative
susceptibility
noted
in
either
the
rat
or
rabbit
developmental
toxicity
studies
or
in
the
two­
generation
reproduction
study.

4.4
Hazard
Identification
and
Toxicity
Endpoint
Selection
4.4.1
Acute
Reference
Dose
(
aRfD)
­
Females
(
13
to
49
Years
of
Age)
No
appropriate
dose
and
endpoint
could
be
identified
for
acute
dietary
exposure
in
Females
(
13­
49).
No
pre­
natal
or
fetal
toxicity
attributable
to
a
single
oral
dose
was
observed
in
the
developmental
toxicity
studies
in
rats
or
rabbits
or
in
the
two­
generation
reproduction
study
in
rats.
In
the
main
developmental
rabbit
study,
offspring
toxicity
was
not
observed
up
to
the
highest
dose
tested.
Although
abortions
were
seen
in
the
main
study,
the
percentage
of
animals
that
aborted
fell
within
the
range
of
the
historical
control
data.
Higher
dose
levels
were
tested
in
the
preliminary
developmental
rabbit
studies.
In
these
studies,
dose­
dependent
increases
in
late­
Page
24
of
48
term
abortions
(
and
maternal
mortality)
were
observed
in
animals
following
severe
changes
in
maternal
body
weights
associated
with
decreased
food
consumption.
Maternal
toxicity,
indicated
by
the
decreases
in
food
consumption
and
body
weights,
was
seen
following
multiple
doses
of
kasugamycin
hydrochloride
hydrate.
In
the
first
preliminary
rabbit
developmental
toxicity
study,
in
which
all
treated
animals
were
sacrificed
in
extremis,
all
of
the
animals
survived
for
at
least
8
days
of
dosing
up
to
the
limit
dose
(
1000
mg/
kg/
day);
there
were
no
abortions
in
this
study.
Because
the
abortions
only
occurred
after
maternal
toxicity
developed
following
multiple
doses
of
kasugamycin
hydrochloride
hydrate,
the
abortions
seen
in
the
rabbit
developmental
toxicity
study
are
not
attributable
to
a
single
oral
dose.

4.4.2
Acute
Reference
Dose
(
aRfD)
­
General
Population
No
appropriate
dose
and
endpoint
could
be
identified
for
acute
dietary
exposure
in
the
general
population,
including
infants
and
children.
No
neurotoxic
effects,
maternal
toxicity
following
1
or
2
doses
in
the
developmental
studies,
or
other
effects
were
seen
following
a
single
oral
dose
that
could
be
relevant
for
this
population
group.

4.4.3
Chronic
Reference
Dose
(
cRfD)
Study
Selected:
Combined
chronic
toxicity/
carcinogenicity
study
in
rats.
MRID
Number:
45910024.
Executive
Summary:
In
a
combined
chronic
toxicity/
carcinogenicity
study
(
MRID
#
45910024),
kasugamycin
hydrochloride
hydrate
(
67.1%
ai,
dose
levels
adjusted
for
purity;
Lot
#
KP­
570)
was
administered
to
70
Wistar
rats/
sex/
dose
in
the
diet
at
dose
levels
of
0,
30,
300,
or
3000
ppm
(
equivalent
to
0/
0,
1.1/
1.4,
11.3/
13.4,
and
115.9/
139.8
mg/
kg
bw/
day
in
males/
females)
for
up
to
2
years.
Ten
rats/
sex/
dose
were
sacrificed
at
Weeks
26
and
52,
and
the
remaining
survivors
were
sacrificed
at
Week
104.
No
treatment­
related
effects
were
observed
on
mortality,
clinical
signs,
food
consumption
or
efficiency,
water
consumption,
ophthalmologic
examination,
hematology,
clinical
chemistry,
or
urinalysis.
Adverse
findings
were
confined
to
the
testes
in
the
3000
ppm
males.
Non­
statistically
significant
increased
incidences
of
testicular
softening
and
atrophy
were
observed
grossly
at
interim
sacrifice
on
Week
26
(
n=
10;
20
to
30%
treated
vs
0%
controls),
at
interim
sacrifice
on
Week
52
(
n=
10;
30%
treated
vs
0%
controls),
and
when
all
males
were
combined
on
Week
104
(
n=
69­
70;
19
to
20%
treated
vs
11%
controls).
Non­
statistically
significant
increased
incidences
of
unilateral
or
bilateral
testicular
tubular
atrophy
were
observed
as
follows:
(
1)
slight
to
moderate
at
Week
26
(
30%
treated
vs
0%
controls),
(
2)
severe
at
Week
52
(
30%
treated
vs
0%
controls),
and
(
3)
slight
to
severe
when
all
males
were
combined
(
n=
69­
70;
25%
treated
vs
14%
controls).
Severity
of
the
tubular
atrophy
in
all
males
was
also
dependent
on
the
dose.
Additionally,
in
a
concurrently
submitted
2­
generation
reproduction
study
in
rats
(
MRID
#
45910023),
the
testes
were
adversely
affected
in
both
the
parents
and
offspring,
with
signs
including
testicular
atrophy/
degeneration,
small
testes,
and
seminiferous
tubule
atrophy.
Although
gross
and
microscopic
testicular
atrophy
is
common
in
old
rats,
increased
atrophy
at
the
interim
sacrifices
suggests
a
treatment­
related
effect.
Body
weights
were
slightly
and
transiently
decreased
in
the
3000
ppm
females,
resulting
in
minor
decreases
in
cumulative
body
weight
gains
during
Weeks
0
to
13
and
0
to
52.
Overall
body
Page
25
of
48
weight
gains
(
Weeks
0
to
104)
were
similar
to
controls.
As
the
effects
on
body
weight
gains
were
slight
and
only
occurred
during
the
first
year
of
the
study,
these
findings
were
not
considered
adverse.
Additionally
at
3000
ppm,
several
findings
were
observed
in
the
kidney,
lungs,
nose,
and
liver
that
were
considered
to
be
unrelated
to
treatment.
Relative
(
to
body)
kidney
weights
were
increased
slightly
in
both
sexes
at
Week
104.
Increased
incidences
of
lipofuscin
deposition
of
proximal
tubular
cells
(
usually
bilateral)
of
the
kidneys
were
observed
as
follows:
(
1)
in
the
males
at
Week
26,
(
2)
in
both
sexes
at
Week
52,
and
(
3)
in
both
sexes
when
all
animals
were
combined.
There
was
no
indication
that
the
etiology
of
the
lipofuscin
deposition
was
related
to
treatment.
Without
supporting
clinical
chemistry,
urinalysis,
or
gross/
microscopic
pathology
to
corroborate
nephrotoxicity,
the
slight
increases
in
relative
kidney
weight
were
considered
to
be
incidental.
Increased
incidences
of
foam
cell
aggregation
in
the
lungs
were
observed
in
the
males
at
Week
52,
and
in
both
sexes
when
all
animals
were
combined.
No
further
evidence
of
toxicity
to
the
lung
was
observed
during
gross
or
microscopic
pathology;
therefore,
these
findings
were
considered
to
be
incidental.
An
increased
incidence
of
slight
to
moderate
rhinitis
vs
slight
in
controls
was
observed
when
all
treated
males
were
combined.
Considering
the
nature
of
the
lesion,
and
slight
increase
in
frequency
and
severity,
this
minor
condition
was
not
considered
adverse.
Total
cholesterol
was
slightly
decreased
in
the
3000
ppm
females
after
52,
78,
and
104
weeks
of
treatment.
However,
this
slight
decrease
may
be
within
the
range
of
natural
variation
(
historical
control
data
were
not
provided).
When
all
animals
were
combined,
acidophilic
cell
foci
of
cellular
alteration
in
the
liver
was
observed
in
the
females
at
300
ppm
or
higher;
however,
the
effect
was
not
clearly
related
to
dose
and
was
considered
equivocal.
Additionally
in
the
females,
increased
incidences
of
slight
hepatocellular
atrophy
were
observed
at
Week
52,
and
when
all
females
were
combined.
Without
any
evidence
of
a
clear
hepatotoxic
effect,
the
hepatocellular
atrophy
was
considered
to
be
incidental.
The
LOAEL
is
116
mg/
kg/
day
in
males,
based
on
increased
testicular
softening
and
atrophy;
the
LOAEL
in
females
was
not
observed.
The
NOAEL
is
11.3/
140
mg/
kg/
day
in
males/
females.
At
the
doses
tested,
there
was
not
a
treatment­
related
increase
in
tumor
incidence
when
compared
to
controls.
Dosing
was
considered
adequate
based
on
increased
lesions
in
the
testes.
This
study
is
classified
acceptable/
guideline
and
satisfies
the
guideline
requirements
for
a
combined
chronic
toxicity/
carcinogenicity
study
(
OPPTS
870.4300;
OECD
453)
in
the
rat.
Dose
and
Endpoint
for
Establishing
cRfD:
11.3
mg/
kg/
day
(
NOAEL)
based
on
increased
testicular
softening
and
atrophy
in
males
at
116
mg/
kg/
day
(
LOAEL).
Uncertainty
Factor(
s):
100
(
10X
for
interspecies
extrapolation
and
10X
for
intraspecies
variations).
Comments
about
Study/
Endpoint/
Uncertainty
Factor:
Although
they
did
not
reach
statistical
significance,
the
testicular
effects
observed
in
high­
dose
male
rats
in
the
combined
chronic/
carcinogenicity
study
were
selected
as
the
basis
for
the
study
LOAEL,
as
well
as
for
the
chronic
dietary
endpoint,
because
an
increase
in
the
incidence
and
severity
of
these
effects
were
noted
and
because
similar
effects
were
seen
across
species
in
multiple
studies.
In
the
chronic
rat
study,
the
incidences
of
testicular
tubular
atrophy
observed
histopathologically
at
week
26,
week
52,
and
when
males
at
all
time
points
were
combined
were
30%,
30%,
and
25%,
respectively,
of
Page
26
of
48
the
high­
dose
group
and
0%,
0%,
and
14%,
respectively,
of
the
controls.
Additionally,
there
was
an
increase
in
severity
of
the
histopathological
testicular
tubular
atrophy.
When
all
males
were
combined,
9%
of
the
controls
and
6%
of
the
high­
dose
group
had
slight
testicular
atrophy,
while
6%
of
the
controls
and
19%
of
the
high­
dose
males
had
moderate
or
severe
testicular
atrophy.
Testicular
tubular
atrophy
and
gross
testicular
changes
were
also
observed
in
rats
in
the
twogeneration
reproduction
study;
see
the
next
paragraph
for
more
details.
In
the
subchronic
mouse
study,
7/
10
and
4/
10
of
the
high­
dose
males
had
tubular
dilatation
and
degeneration,
respectively,
compared
to
0/
10
and
0/
10
of
the
controls.
Although
these
testicular
effects
were
not
replicated
in
the
chronic
mouse
study,
the
highest
dose
tested
in
the
chronic
mouse
study
was
half
the
value
of
the
LOAEL
for
the
subchronic
mouse
study.
In
the
subchronic
dog
study,
1/
4
of
the
high­
dose
dogs
had
hypospermia
compared
to
0/
4
of
the
other
dose
groups.
In
the
chronic
dog
study,
2/
4
of
the
high­
dose
males
had
testicular
degeneration
and
3/
4
had
chronic
inflammation;
only
1/
4
of
the
controls
had
degeneration
and
0/
4
had
inflammation.
The
two­
generation
reproduction
study
in
rats
(
MRID
#
45910023)
was
considered
a
cocritical
study.
The
executive
summary
for
this
study
is
found
in
Section
4.2.4.
In
the
twogeneration
reproduction
study,
the
LOAEL
for
parental
toxicity
was
1000
ppm
(
equivalent
to
70.3/
82.9
mg/
kg/
day
[
M/
F]),
based
on
decreased
body
weights
and
body
weight
gains
seen
in
the
P
generation
males.
The
NOAEL
for
this
study
is
200
ppm
(
equivalent
to
13.7/
16.2
mg/
kg/
day
[
M/
F]).
Testicular
effects
similar
to
those
found
in
the
combined
chronic/
carcinogenicity
study
in
rats
were
also
observed
in
this
study
at
a
dose
greater
than
the
reproduction
study
LOAEL.
At
425
mg/
kg/
day,
testicular
tubular
atrophy
was
seen
in
12%
of
the
parental
males
and
in
63%
of
the
F1
generation
males,
compared
to
none
of
the
control
P
or
F1
males.
Grossly,
at
this
same
dose,
38%
of
the
F1
generation
had
small
testes.
The
F1
male
fertility
index
decreased
to
38%
at
425
mg/
kg/
day
compared
to
88%
in
controls.
The
NOAEL
and
LOAEL
from
the
subchronic
dog
study
(
10.6
and
106.0
mg/
kg/
day,
respectively)
are
slightly
lower
than
the
NOAEL
and
LOAEL
from
the
combined
chronic/
carcinogenicity
study
in
rats
(
11.3
and
116
mg/
kg/
day,
respectively).
However,
this
study
was
not
selected
to
establish
the
cRfD
because
the
effects
used
as
a
basis
for
the
LOAEL
in
the
subchronic
dog
study
(
ie,
excessive
salivation,
swollen
mouth
with
thickened
skin,
tongue
erosions
and
ulcers,
histopathological
lesions
of
the
tongue,
and
few
feces)
were
not
seen
at
similar
doses
tested
in
the
chronic
dog
study.
Although
the
treatment­
related
effects
in
the
subchronic
study
were
not
used
to
establish
the
cRfD,
the
selected
NOAEL
is
protective
of
these
effects.

Chronic
RfD
=
11.3
mg/
kg/
day
(
NOAEL)
=
0.113
mg/
kg/
day
100
(
UF)

4.4.4
Incidental
Oral
Exposure
(
Short
and
Intermediate
Term)
Because
no
proposed
uses
would
result
in
residential
exposure
in
the
US,
no
incidental
oral
exposure
assessment
is
required
at
this
time.

4.4.5
Dermal
Absorption
Because
no
proposed
uses
would
result
in
residential
or
occupational
exposure
in
the
US,
no
dermal
exposure
assessment
is
required
at
this
time.
As
such,
dermal
absorption
and
dermal
toxicity
studies
are
currently
not
required,
and
a
dermal
absorption
factor
is
not
currently
needed.
Page
27
of
48
4.4.6
Dermal
Exposure
(
Short,
Intermediate
and
Long
Term)
Because
no
proposed
uses
would
result
in
residential
or
occupational
exposure
in
the
US,
no
dermal
exposure
assessment
is
required
at
this
time.

4.4.7
Inhalation
Exposure
(
Short,
Intermediate
and
Long
Term)
Because
no
proposed
uses
would
result
in
residential
or
occupational
exposure
in
the
US,
no
inhalation
exposure
assessment
is
required
at
this
time.

4.4.8
Margins
of
Exposure
Because
no
proposed
uses
would
result
in
residential
or
occupational
exposure
in
the
US,
no
incidental
oral,
dermal,
or
inhalation
exposure
assessments
are
required
at
this
time.
Therefore,
it
is
not
currently
necessary
to
set
levels
of
concern
for
margins
of
exposure.

4.4.9
Recommendation
for
Aggregate
Exposure
Risk
Assessments
Because
no
proposed
uses
would
result
in
residential
exposure
in
the
US,
an
aggregate
exposure
assessment
including
any
exposures
other
than
dietary
(
food
only)
is
not
required
at
this
time.
Based
on
the
proposed
tolerances
on
imported
commodities,
exposure
to
kasugamycin
will
not
occur
via
the
drinking
water,
incidental
oral,
dermal,
or
inhalation
pathways.

4.4.10
Classification
of
Carcinogenic
Potential
The
risk
assessment
team
has
classified
kasugamycin
as
"
not
likely
to
be
carcinogenic
to
humans."
No
oncogenic
potential
was
noted
in
the
mouse
oncogenicity
or
in
the
rat
combined
chronic/
carcinogenicity
studies;
additionally,
no
mutagenic
potential
was
noted
in
any
of
the
five
mutagenicity
studies.

TABLE
4.4
Summary
of
Toxicological
Doses
and
Endpoints
for
Kasugamycin
to
be
Used
in
Human
Health
Risk
Assessments.

Exposure
Scenario
Dose
Used
in
Risk
Assessment
and
UF
1
Special
FQPA
SF
2
and
Level
of
Concern
for
Risk
Assessment
Study
and
Toxicological
Effects
Acute
Dietary
(
females
13
to
49
years
of
age)
None
None
Not
selected
No
appropriate
dose
and
endpoint
could
be
identified
for
these
population
groups.

Acute
Dietary
(
general
population
including
infants
and
children)
None
None
Not
selected
No
appropriate
dose
and
endpoint
could
be
identified
for
these
population
groups.

Chronic
Dietary
(
all
populations)
NOAEL
=
11.3
mg/
kg/
day
UF
=
100
Chronic
RfD
=
0.113
mg/
kg/
day
FQPA
SF
=
1
cPAD
=
chronic
RfD
FQPA
SF
=
0.113
mg/
kg/
day
Combined
chronic
toxicity/
oncogenicity
study
in
rats
LOAEL
=
116
mg/
kg/
day
based
on
increased
testicular
softening
and
atrophy.
TABLE
4.4
Summary
of
Toxicological
Doses
and
Endpoints
for
Kasugamycin
to
be
Used
in
Human
Health
Risk
Assessments.

Exposure
Scenario
Dose
Used
in
Risk
Assessment
and
UF
1
Special
FQPA
SF
2
and
Level
of
Concern
for
Risk
Assessment
Study
and
Toxicological
Effects
Page
28
of
48
Incidental
Oral
(
all
durations)
None
None
Not
selected
Tolerance
on
imported
commodities
­
no
proposed
uses
would
result
in
residential
exposure
in
the
US.

Dermal
(
all
durations)
None
None
Not
selected
Tolerance
on
imported
commodities
­
no
proposed
uses
would
result
in
residential
or
occupational
exposure
in
the
US.

Inhalation
(
all
durations)
None
None
Not
selected
Tolerance
on
imported
commodities
­
no
proposed
uses
would
result
in
residential
or
occupational
exposure
in
the
US.

Cancer
(
oral,
dermal,
inhalation)
Classification:
No
oncogenic
potential
was
noted
in
the
mouse
oncogenicity
or
in
the
rat
combined
chronic/
carcinogenicity
studies;
additionally,
no
mutagenic
potential
was
noted
in
any
of
the
five
mutagenicity
studies.
Classification
of
kasugamycin
is
"
not
likely
to
be
carcinogenic
to
humans".

1.
UF
=
Uncertainty
Factor.
2.
FQPA
SF
=
special
FQPA
Safety
Factor
(
refer
to
Section
4.5).
3.
NOAEL
=
No
Observed
Adverse
Effect
Level.
4.
RfD
=
Reference
Dose.
5.
PAD
=
Population­
Adjusted
Dose
(
a
=
acute,
c
=
chronic).
6.
LOAEL
=
Lowest
Observed
Adverse
Effect
Level.

4.5
Special
FQPA
Safety
Factor
Based
on
the
hazard
and
exposure
data,
the
kasugamycin
risk
assessment
team
has
recommended
that
the
special
FQPA
SF
be
reduced
to
1X
because
there
are
no/
low
concerns
and
no
residual
uncertainties
with
regard
to
pre­
and/
or
post­
natal
toxicity.
This
recommendation
is
based
on
the
following:
(
1)
there
are
no
data
gaps
for
the
assessment
of
the
effects
of
kasugamycin
following
in
utero
and/
or
post­
natal
exposure;
a
developmental
neurotoxicity
study
is
not
required;
(
2)
there
is
no
indication
of
increased
quantitative
or
qualitative
susceptibility
of
rats
or
rabbits
to
in
utero
and/
or
post­
natal
exposure
to
kasugamycin;
(
3)
the
acute
and
chronic
dietary
food
exposure
assessments
utilize
proposed
tolerance
level
or
higher
residues
and
100%
crop
treated
information
for
all
commodities;
by
using
these
screening­
level
assessments,
acute
and
chronic
exposures/
risks
will
not
be
underestimated;
and
(
4)
there
are
no
existing
or
proposed
residential
uses
for
kasugamycin
at
this
time.

4.6
Endocrine
Disruption
EPA
is
required
under
the
FFDCA,
as
amended
by
FQPA,
to
develop
a
screening
program
Page
29
of
48
to
determine
whether
certain
substances
(
including
all
pesticide
active
and
other
ingredients)
"
may
have
an
effect
in
humans
that
is
similar
to
an
effect
produced
by
a
naturally
occurring
estrogen,
or
other
such
endocrine
effects
as
the
Administrator
may
designate."
Following
recommendations
of
its
Endocrine
Disruptor
and
Testing
Advisory
Committee
(
EDSTAC),
EPA
determined
that
there
was
a
scientific
basis
for
including,
as
part
of
the
program,
the
androgen
and
thyroid
hormone
systems,
in
addition
to
the
estrogen
hormone
system.
EPA
also
adopted
EDSTAC's
recommendation
that
the
Program
include
evaluations
of
potential
effects
in
wildlife.
For
pesticide
chemicals,
EPA
will
use
FIFRA
and,
to
the
extent
that
effects
in
wildlife
may
help
determine
whether
a
substance
may
have
an
effect
in
humans,
FFDCA
authority
to
require
the
wildlife
evaluations.
As
the
science
develops
and
resources
allow,
screening
of
additional
hormone
systems
may
be
added
to
the
Endocrine
Disruptor
Screening
Program
(
EDSP).
For
kasugamycin,
effects
which
indicate
potential
endocrine
disruption
include
changes
in
the
testes
(
eg,
lesions,
softening,
atrophy,
and
degeneration)
in
rats
and
mice.
Decreased
fertility,
decreased
fecundity,
and
an
increased
pre­
coital
interval
during
the
mating
period
were
seen
in
rats;
these
effects
may
be
reflections
of
the
changes
in
the
testes.
When
additional
appropriate
screening
and/
or
testing
protocols
being
considered
under
the
Agency's
EDSP
have
been
developed,
kasugamycin
may
be
subjected
to
further
screening
and/
or
testing
to
better
characterize
effects
related
to
endocrine
disruption.

5.0
Public
Health
Data
Kasugamycin
is
a
new
active
ingredient
and,
as
such,
no
public
health
data
are
currently
available.
Kasugamycin
operates
via
a
mode
of
action
different
from
that
of
the
other
aminoglycoside
antibiotics
such
as
streptomycin.
Because
kasugamycin
is
active
only
against
phytopathogenic
fungi
and
bacteria,
it
has
never
been
employed
as
a
human
or
veterinary­
use
antibiotic.
HED
is
aware
that
FDA
and
CDC
have
concerns
regarding
the
potential
for
antibiotics
to
induce
bacterial
resistance
arising
from
their
use
as
pesticides.
HED
has
met
with
these
agencies
recently
to
discuss
resistance
issues
and
an
ongoing
dialogue
is
anticipated.
HED's
level
of
concern
is
low
regarding
development
of
resistance
(
associated
with
kasugamycin's
use
as
a
fungicide)
arising
from
tolerances
for
kasugamycin
on
imported
fruiting
vegetables
because:
(
1)
proposed
use
rates
for
kasugamycin
are
low,
and
residues
following
its
application
are
either
very
low
or
non­
detectable,
(
2)
the
proposed
uses
are
only
on
imported
fruiting
vegetables,
with
no
proposed
domestic
uses,
and
(
3)
there
are
no
human
or
veterinary
uses
of
kasugamycin
as
an
antibiotic.

6.0
Exposure
Characterization/
Assessment
6.1
Dietary
Exposure/
Risk
Pathway
6.1.1
Residue
Profile
The
submitted
crop
field
trial
data
for
peppers
and
tomatoes
are
adequate
to
support
the
proposed
uses
on
fruiting
vegetables.
Residues
of
kasugamycin
were
below
the
method
LOQ
(
less
than
0.040
ppm)
in/
on
all
samples
of
peppers
and
tomatoes
harvested
1
day
following
the
Page
30
of
48
last
of
three
foliar
broadcast
applications
of
the
Kasumin
®
2L
liquid
formulation
at
total
seasonal
rates
of
0.052
to
0.054
lb
ai/
A.
In
residue
decline
trials,
residues
were
non­
detectable
in/
on
all
samples
of
peppers
and
tomatoes
collected
0,
1,
3,
and
7
days
after
final
application;
therefore,
it
is
not
possible
to
make
a
conclusion
concerning
whether
kasugamycin
residues
decline
in
fruiting
vegetables
with
increasing
sampling
intervals
following
treatment.
The
tomato
field
trials
included
both
processing
and
fresh
market
tomato
varieties;
however,
no
small
varieties
of
tomatoes
(
cherry
tomatoes)
were
included.
The
studies
are
supported
by
adequate
storage
stability
data.
Processing
studies
on
tomato,
for
the
processed
commodities
tomato
puree
and
tomato
paste
(
residue
data
on
paste
cover
tomato
juice),
were
not
conducted;
a
request
for
waiver
of
the
requirement
for
tomato
processing
data
was
submitted.
Based
on
the
results
of
field
trial
data
at
the
5X
exaggerated
rate,
residue
data
on
small
tomato
varieties
and
a
tomato
processing
study
are
not
required.
The
proposed
residue
analytical
method
is
adequate
for
data
collection;
however,
radiovalidation
data
were
not
submitted.
The
method
was
submitted
to
ACL/
BEAD
for
a
PMV
trial
(
PMV
Request
Memo,
D313672,
William
T.
Drew,
3/
1/
2005),
which
it
subsequently
failed
(
PMV
Results
Memo,
D313673,
Patricia
G.
Schermerhorn,
7/
13/
2005).
The
method
includes
instructions
for
the
analysis
of
tomato
and
pepper
samples.
Briefly,
macerated
or
ground
frozen
samples
are
extracted
twice
with
methanol/
water
(
7:
3
vol/
vol)
at
pH
4.
The
extracts
are
filtered,
combined,
and
concentrated,
then
filtered
through
diatomaceous
earth
to
remove
precipitates.
The
filtered
concentrate
is
diluted
with
water
and
purified
using
two
separate
ion­
exchange
columns.
The
resulting
purified
extract
is
concentrated
before
analysis
via
reverse­
phase,
ionpairing
HPLC/
UV
analysis.
The
validated
LOQ
is
0.040
ppm,
determined
as
the
lower
limit
of
method
verification
(
LLMV),
and
the
calculated
limit
of
detection
(
LOD)
is
0.013
ppm.
The
method
does
not
include
a
confirmatory
method,
and
no
interference
study
was
conducted.

6.1.2
Acute
and
Chronic
Dietary
Exposure
and
Risk
The
dietary
exposure
assessment,
entitled
Kasugamycin
Chronic
Dietary
Exposure
Assessment
for
the
Import
Tolerance
on
Fruiting
Vegetables,
was
conducted
by
Douglas
A.
Dotson,
5/
9/
2005,
DP
Barcode
D315531.
A
chronic
dietary
risk
assessment
was
conducted
for
the
new
active
ingredient
kasugamycin
using
the
Dietary
Exposure
Evaluation
Model
(
DEEM­
FCID,
Version
2.03),
which
uses
food
consumption
data
from
the
USDA's
Continuing
Surveys
of
Food
Intakes
by
Individuals
(
CSFII)
from
1994
to
1996
and
1998.
The
analysis
was
performed
to
support
requests
for
tolerances
on
imported
fruiting
vegetables
(
Crop
Group
8).
Acute
Dietary
Exposure:
An
acute
dietary
exposure
analysis
was
not
performed
for
kasugamycin,
because
no
appropriate
dose
or
endpoint
could
be
identified
for
acute
dietary
exposure
in
the
general
population
or
any
of
the
population
subgroups.
Chronic
Dietary
Exposure
Results
and
Characterization:
A
conservative
chronic
dietary
exposure
analysis
was
performed
for
kasugamycin.
The
analysis
is
based
on
tolerance­
level
residues
(
modified
by
DEEM
default
processing
factors)
and
the
assumption
that
100%
of
the
crop
will
be
treated.
The
risk
estimates
for
all
population
subgroups
are
below
HED's
level
of
concern.
The
risk
estimate
for
the
general
US
population
is
less
than
1%
of
the
cPAD.
The
most
highly
exposed
population
subgroup
is
Children
(
1­
2),
which
utilizes
less
than
1%
of
the
cPAD.
Page
31
of
48
TABLE
6.1
Summary
of
Dietary
Exposure
and
Risk
for
Kasugamycin.

Population
Subgroup
Acute
Dietary
Chronic
Dietary
1
aPAD
(
mg/
kg)
Exposure
(
mg/
kg/
day)
%
aPAD
cPAD
(
mg/
kg/
day)
Exposure
(
mg/
kg/
day)
2
%
cPAD
General
US
Population
Not
applicable.
0.113
0.000073
<
1
All
Infants
<
1
year
0.113
0.000040
<
1
Children
1­
2
years
0.113
0.00018
<
1
Children
3­
5
years
0.113
0.00016
<
1
Children
6­
12
years
0.113
0.00011
<
1
Youths
13­
19
years
0.113
0.000075
<
1
Adults
20­
49
years
0.113
0.000063
<
1
Adults
50+
years
0.113
0.000050
<
1
Females
13­
49
years
0.113
0.000060
<
1
1.
Values
for
the
population
with
the
highest
risk
for
each
type
of
risk
assessment
are
bolded.
2.
Reported
to
2
significant
figures.

6.2
Water
Exposure/
Risk
Pathway
Since
kasugamycin
is
proposed
for
use
only
on
imported
fruiting
vegetable
commodities,
the
sole
anticipated
exposure
route
for
the
US
population
is
via
dietary
(
food)
exposure.
With
no
proposed
US
registration,
there
is
no
expectation
that
kasugamycin
residues
would
occur
in
surface
or
ground
water
sources
of
drinking
water.

6.3
Residential
(
Non­
Occupational)
Exposure/
Risk
Pathway
Since
kasugamycin
is
proposed
for
use
only
on
imported
fruiting
vegetable
commodities,
the
sole
anticipated
exposure
route
for
the
US
population
is
via
dietary
(
food
only)
exposure.
With
no
proposed
US
registration,
kasugamycin
is
not
intended
for
use
in
public
or
residential
settings.
Therefore,
residential
exposure
is
not
expected
and
no
residential
risk
assessment
was
performed.

7.0
Aggregate
Risk
Assessments
and
Risk
Characterization
Since
kasugamycin
is
proposed
for
use
only
on
imported
fruiting
vegetable
commodities,
the
sole
anticipated
exposure
route
for
the
US
population
is
via
dietary
(
food
only)
exposure.
With
no
proposed
US
registration,
there
is
no
expectation
that
exposure
to
kasugamycin
residues
would
occur
via
water
consumption
or
residential
use.
Therefore,
aggregate
exposure
is
not
expected
and
no
aggregate
risk
assessment
was
performed.

8.0
Cumulative
Risk
Characterization/
Assessment
Unlike
other
pesticides
for
which
EPA
has
followed
a
cumulative
risk
approach
based
on
a
common
mechanism
of
toxicity,
EPA
has
not
made
a
common
mechanism
of
toxicity
finding
as
to
kasugamycin
and
any
other
substances,
and
kasugamycin
does
not
appear
to
produce
a
toxic
metabolite
produced
by
other
substances.
For
the
purposes
of
this
tolerance
action,
therefore,
Page
32
of
48
EPA
has
not
assumed
that
kasugamycin
has
a
common
mechanism
of
toxicity
with
other
substances.
For
information
regarding
EPA's
efforts
to
determine
which
chemicals
have
a
common
mechanism
of
toxicity
and
to
evaluate
the
cumulative
effects
of
such
chemicals,
see
the
policy
statements
released
by
EPA's
Office
of
Pesticide
Programs
(
OPP)
concerning
common
mechanism
determinations
and
procedures
for
cumulating
effects
from
substances
found
to
have
a
common
mechanism
on
EPA's
website
at
http://
www.
epa.
gov/
pesticides/
cumulative/.

9.0
Occupational
Exposure/
Risk
Pathway
Since
kasugamycin
is
proposed
for
use
only
on
imported
fruiting
vegetable
commodities,
the
sole
anticipated
exposure
route
for
the
US
population
is
via
dietary
(
food)
exposure.
With
no
proposed
US
registration,
there
is
no
expectation
that
exposure
to
kasugamycin
residues
would
occur
via
occupational
use.
Therefore,
no
occupational
risk
assessment
was
performed.

10.0
Data
Needs
and
Label
Requirements
10.1
Toxicology
No
additional
toxicological
studies
are
required
for
kasugamycin
at
this
time.

10.2
Residue
Chemistry
HED
has
examined
the
residue
chemistry
database
for
kasugamycin
and
recommends
against
the
establishment
of
tolerances
for
residues
of
this
fungicide
in/
on
imported
fruiting
vegetables
until
the
data
deficiencies
listed
below
(
OPPTS
Residue
Chemistry
Test
Guideline
860.1340)
have
been
resolved.
The
fact
that
an
adequate
analytical
method
for
tolerance
enforcement
is
unavailable
does
not
impact
this
risk
assessment
regarding
risk
characterization.
860.1340
Residue
Analytical
Method
(
Plant
Commodities)
(
1)
The
proposed
enforcement
method
should
be
modified
to
include
a
confirmatory
analysis
method;
alternatively,
the
petitioner
may
submit
an
interference
study
with
all
pesticides
for
which
tolerances
on
tomatoes
and/
or
peppers
have
been
established.
(
2)
The
method
has
failed
a
PMV
trial
(
PMV
Results
Memo,
D313673,
Patricia
G.
Schermerhorn,
7/
13/
2005),
and
should
be
modified
to
include
the
revisions
recommended
by
ACL.
860.1550
Proposed
Tolerances
The
available
crop
field
trial
data
support
a
tolerance
for
residues
of
kasugamycin
per
se
in/
on
imported
fruiting
vegetable
commodities
(
Crop
Group
8)
at
0.04
ppm,
the
method's
LOQ.
Based
on
the
submitted
field
trial
data,
the
following
use
pattern
would
be
supported
for
fruiting
vegetables:
application
of
the
Kasumin
®
2L
formulation
as
a
foliar
broadcast
spray
in
a
minimum
spray
volume
of
30
GPA
using
ground
equipment,
with
no
spray
adjuvants,
a
minimum
RTI
of
3
days,
and
a
PHI
of
1
day.
HED
recommends
that
the
petitioner's
request
for
waiver
of
tomato
processing
data
be
granted;
a
tomato
processing
study
is
not
required.
The
exaggerated­
rate
(
5X)
tomato
field
trial
data
indicate
that
separate
tolerances
for
tomato
juice,
tomato
puree
and
tomato
paste
are
not
required.
The
proposed
tolerances
should
be
revised
to
reflect
the
tolerance
as
recommended
by
HED
and
the
correct
commodity
definition
as
specified
in
Table
10.2,
below.
The
petitioner
should
submit
a
revised
Section
F
for
PP#
3E6579
to
reflect
these
changes.
Page
33
of
48
TABLE
10.2
Tolerance
Summary
for
Kasugamycin.

Commodity
Proposed
Tolerance
(
ppm)
Recommended
Tolerance
(
ppm)
Comments/
Correct
Commodity
Definition
Fruiting
Vegetables
(
Crop
Group
8)
0.04
0.04
Vegetable,
fruiting,
group
8
Tomato
Juice
0.06
None
Residues
are
not
expected
to
exceed
the
tolerance
on
the
RAC.
Tomato
Puree
0.06
Tomato
Paste
0.25
References:
1.
Kasugamycin
Chronic
Dietary
Exposure
Assessment
for
the
Import
Tolerance
on
Fruiting
Vegetables.
D315531,
Douglas
Dotson,
5/
9/
2005.
2.
Kasugamycin.
Tolerance
Petition
Requesting
Food
Use
of
the
Fungicide
Kasugamycin
on
Imported
Fruiting
Vegetables
(
Crop
Group
8).
Summary
of
Analytical
Chemistry
and
Residue
Data.
Petition
Number
3E6579.
D308573,
William
T.
Drew,
8/
17/
2005.

Appendices
A­
1.0
TOXICOLOGY
DATA
REQUIREMENTS
The
requirements
(
40
CFR
158.340)
for
food
use
(
tolerances
on
imported
commodities
only)
of
kasugamycin
are
in
Appendix
Table
1.
Use
of
the
new
guideline
numbers
does
not
imply
that
the
new
(
1998)
guideline
protocols
were
used.

APPENDIX
TABLE
1
Toxicology
Data
Requirements.

Test
Technical
Required
Satisfied
870.1100
Acute
Oral
Toxicity
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.1200
Acute
Dermal
Toxicity
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.1300
Acute
Inhalation
Toxicity
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.2400
Primary
Eye
Irritation
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.2500
Primary
Dermal
Irritation
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.2600
Dermal
Sensitization
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
870.3100
Oral
Subchronic
(
rodent)
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.3150
Oral
Subchronic
(
nonrodent)
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.3200
21­
Day
Dermal
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.3250
90­
Day
Dermal
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.3465
90­
Day
Inhalation
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
yes
yes
no
no
no
yes
yes
no
no
no
870.3700a
Developmental
Toxicity
(
rodent)
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.3700b
Developmental
Toxicity
(
nonrodent)
.
.
.
.
.
.
.
.
.
.
.
.
.
870.3800
Reproduction
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
yes
yes
yes
yes
yes
yes
APPENDIX
TABLE
1
Toxicology
Data
Requirements.

Test
Technical
Required
Satisfied
Page
34
of
48
870.4100a
Chronic
Toxicity
(
rodent)
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.4100b
Chronic
Toxicity
(
nonrodent)
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.4200a
Oncogenicity
(
rat)
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.4200b
Oncogenicity
(
mouse)
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.4300
Chronic/
Oncogenicity
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
870.5100
Mutagenicity
 
Gene
Mutation
­
bacterial
.
.
.
.
.
.
.
.
.
870.5300
Mutagenicity
 
Gene
Mutation
­
mammalian
.
.
.
.
.
.
.
870.5375
Mutagenicity
 
Structural
Chromosomal
Aberrations
.
870.5xxx
Mutagenicity
 
Other
Genotoxic
Effects
.
.
.
.
.
.
.
.
.
.
.
yes
yes
yes
yes
no*
yes
no*
yes
870.6100a
Acute
Delayed
Neurotox.
(
hen)
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.6100b
90­
Day
Neurotoxicity
(
hen)
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.6200a
Acute
Neurotox.
Screening
Battery
(
rat)
.
.
.
.
.
.
.
.
.
.
870.6200b
90
Day
Neuro.
Screening
Battery
(
rat)
.
.
.
.
.
.
.
.
.
.
.
.
870.6300
Develop.
Neuro
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
no
no
no
no
no
no
no
no
no
no
870.7485
General
Metabolism
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.7600
Dermal
Penetration
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
yes
no
yes
no
*
Although
these
two
mutagenicity
studies
were
not
classified
as
acceptable/
guideline,
the
requirement
for
these
studies
has
been
waived.
See
Section
4.1
for
more
details.

A­
2.0
NON­
CRITICAL
TOXICOLOGY
STUDIES
Executive
summaries
of
the
toxicology
studies
not
used
for
toxicity
endpoint
selection
or
FQPA
assessment
are
as
follows.

A­
2.1
Subchronic
Oral
Toxicity
­
Rat
(
870.3100)
In
a
90­
day
oral
toxicity
study
(
MRID
#
45910020),
kasugamycin
hydrochloride
hydrate
(
64.5%
ai;
Lot
#
KP­
834)
was
administered
to
12
Wistar
rats/
sex/
dose
in
the
diet
at
dose
levels
of
0,
300,
1000,
3000,
or
6000
ppm
(
equivalent
to
0/
0,
17.5/
20.3,
58.2/
69.2,
176.7/
201.0,
and
354.8/
395.5
mg/
kg
bw/
day
in
males/
females,
respectively).
It
was
assumed,
based
on
dose
formulation
concentration
analyses,
that
the
doses
were
adjusted
for
compound
purity;
however,
this
was
not
stated
explicitly.
No
treatment­
related
effects
were
observed
on
mortality,
clinical
signs,
food
consumption,
food
efficiency,
water
consumption,
ophthalmoscopic
examination,
hematology,
clinical
chemistry,
organ
weights,
or
macroscopic
pathology.
There
were
indications
of
a
slight
systemic
toxicity
at
6000
ppm.
At
this
dose,
male
body
weights
were
significantly
lower
(
4­
9%)
than
controls
(
p

0.05)
each
week
starting
on
Week
2
until
sacrifice,
resulting
in
an
11%
decrease
in
overall
(
Weeks
0­
13)
body
weight
gain.
In
the
6000
ppm
females,
body
weights
were
significantly
lower
(
5%)
than
controls
(
p

0.05)
only
on
Week
6;
however,
non­
significant
decreases
in
body
weight
gain
were
observed
throughout
the
Page
35
of
48
study,
resulting
in
a
decrease
of
8%
in
overall
body
weight
gain.
Although
not
considered
treatment­
related,
significant
(
p

0.05)
decreases
in
food
consumption
were
seen
in
both
sexes
during
the
first
week
of
dosing,
indicating
a
possible
palatability
issue;
average
food
consumption
(
Weeks
0­
13)
was
non­
significantly
decreased
by
4­
6%
in
6000
ppm
males
and
females.
An
increased
(
p

0.05)
number
of
epithelial
cells
in
the
urinary
sediment
was
observed
in
the
females.
Urine
pH
was
decreased
in
both
sexes;
this
is
possibly
due
to
the
acidic
properties
of
the
test
compound.
Microscopically,
in
the
6000
ppm
group,
increased
(
p

0.05)
incidences
were
observed
of
eosinophilic
bodies
in
the
proximal
tubular
cells
in
the
kidney
of
slight
severity
in
the
males
(
11/
12
treated
vs.
0/
12
controls)
and
foam
cell
aggregation
in
lungs
(
all
of
slight
severity
except
1
treated
of
moderate
severity,
10/
12
treated
vs.
3/
12
controls).
These
same
lesions
were
found
in
the
3000
ppm
group
in
the
concurrently
submitted
combined
chronic
toxicity
/
carcinogenicity
study
(
MRID
#
45910024),
without
any
additional
treatment­
related
lesions
in
the
organs
to
corroborate
toxicity.
Additionally,
these
lesions
were
graded
slight
(
lowest
grade
reported).
While
related
to
treatment,
the
increased
incidence
of
both
of
these
effects
was
not
considered
biologically
significant.
No
adverse
effects
were
observed
at
doses
equal
to
or
lower
than
3000
ppm.
Although
an
apparent
decrease
in
water
consumption
was
seen
in
the
3000
ppm
males,
mean
water
consumption
data
was
based
on
individual
cage
values,
and
no
baseline
water
consumption
measurements
were
taken
prior
to
dosing.
Therefore,
there
is
not
enough
information
to
make
a
definitive
evaluation
of
a
treatment­
related
effect
on
male
water
consumption.
Decreases
in
hematocrit,
hemoglobin,
and
erythrocyte
counts
at
3000
and
6000
ppm
were
minor
and
not
considered
biologically
significant.
Increased
incidences
of
increased
eosinophilic
bodies
in
the
kidneys
in
males
and
foam
cell
aggregation
in
the
lungs
in
the
females
were
also
not
considered
toxicologically
significant.
The
LOAEL
is
6000
ppm
(
equivalent
to
354.8/
395.5
mg/
kg/
day
in
males/
females),
based
on
decreased
body
weights
and
body
weight
gains
in
both
sexes.
The
NOAEL
is
3000
ppm
(
equivalent
to
176.7/
201.0
mg/
kg/
day
in
males/
females).
This
study
is
classified
acceptable/
guideline
and
satisfies
the
guideline
requirements
for
a
90­
day
oral
toxicity
study
(
OPPTS
870.3100a;
OECD
408)
in
the
rat.

A­
2.2
Subchronic
Oral
Toxicity
­
Mouse
(
870.3100)
In
this
subchronic
oral
toxicity
study
(
MRID
#
45910019),
12
CD­
1
mice/
sex/
dose
were
exposed
to
kasugamycin
hydrochloride
hydrate
(
64.8%
ai;
Batch
#:
KP­
821)
in
the
diet
at
nominal
concentrations
of
0,
300,
1000,
3000,
or
10,000
ppm
(
equivalent
to
0/
0,
41.2/
58.0,
135.4/
170.9,
408.5/
565.6,
and
1559/
1834
mg/
kg/
day
in
males/
females)
for
up
to
14
weeks.
It
was
assumed,
based
on
dose
formulation
concentration
analyses,
that
the
doses
were
adjusted
for
compound
purity;
however,
this
was
not
stated
explicitly.
No
treatment­
related
effects
were
observed
on
food
or
water
consumption,
ophthalmoscopic
examination,
hematology,
clinical
chemistry,
urinalysis,
or
organ
weights.
At

3000
ppm,
generally,
from
Week
4
until
study
termination,
perianal
reddening
was
observed
in
both
sexes.
Increased
incidences
of
minimal
to
marked
active
chronic
inflammation
and
ulceration
of
the
anus
was
observed
in
both
sexes.
Increased
mortality
was
also
observed
in
both
sexes.
Four
of
the
six
decedent
animals
in
the

3000
ppm
groups
were
suffering
from
these
lesions,
and
3
of
these
4
animals
were
killed
because
of
extensive
perianal
skin
lesions.
Minimal
to
severe
basophilia/
hyperplasia
in
the
pars
recta
of
the
kidneys
in
females
was
also
observed.
Page
36
of
48
At
10,000
ppm,
body
weights
were
decreased
generally
throughout
treatment
in
both
sexes,
as
were
overall
(
Weeks
0­
14)
body
weight
gains
and
overall
(
Weeks
1­
14)
food
efficiency.
In
females,
an
increased
incidence
of
emaciated
appearance,
perianal/
perigenital
stain,
and
dark
perianal
regions
were
also
observed.
Increased
incidences
of
testicular
tubular
dilatation,
tubular
degeneration
associated
with
dilatation,
and
spermatoceles
were
observed.
Additionally,
extramedullary
hematopoiesis
in
the
spleen
was
also
observed
in
the
males.
The
LOAEL
is
3000
ppm
(
equivalent
to
408.5/
565.6
mg/
kg/
day
in
males/
females),
based
on
increased
mortality
and
anal
lesions
in
both
sexes,
and
kidney
lesions
in
the
females.
The
NOAEL
is
1000
ppm
(
equivalent
to
135.4/
170.9
mg/
kg/
day
in
males/
females).
This
study
is
classified
as
acceptable/
guideline
and
satisfies
the
guideline
requirements
(
OPPTS
870.3100a;
OECD
408)
for
a
subchronic
oral
toxicity
study
in
the
mouse.

A­
2.3
Subchronic
Oral
Toxicity
­
Dog
(
870.3150)
In
a
subchronic
oral
toxicity
study
(
MRID
#
46030302),
kasugamycin
hydrochloride
hydrate
(
80.6%
ai;
dose
levels
adjusted
for
purity;
Lot
No.:
KP­
913)
was
administered
to
4
beagle
dogs/
sex/
group
in
the
diet
at
dose
levels
of
0,
300,
3000,
or
6000/
4500
ppm
(
equivalent
to
0/
0,
11/
11,
106/
108,
and
182/
179
mg/
kg/
day
[
M/
F],
respectively)
for
13
weeks.
No
treatment­
related
effects
were
observed
on
mortality,
ophthalmoscopy,
hematology,
urinalysis,
or
organ
weight.
At

3000
ppm,
few
feces,
swollen
mouth,
excessive
salivation,
and
thickened
skin
at
the
commissure
of
the
mouth
were
observed
in
both
sexes.
Increased
incidences
of
clinically
observed
tongue
erosions/
ulcerations
were
observed
in
both
sexes.
Histopathological
tongue
lesions,
including
moderate
to
severe
atrophy
of
the
dorsal
epithelium
and
moderate
to
severe
loss
of
epithelial
papillae
of
the
dorsal
surface,
were
observed
in
both
sexes;
and
moderate
to
severe
tongue
ulcerations
were
observed
in
males.
In
the
high
dose
group
initially
at
6000
ppm,
tongue
erosions/
ulcerations
were
clinically
observed
and
food
consumption
during
Weeks
1
through
5
was
reduced
(­
6
to
­
45%)
in
both
sexes.
Consequently,
these
animals
were
offered
control
diet
between
Weeks
6
(
Day
41)
and
8
(
Day
50)
and
started
on
a
reduced
dietary
dose
of
4500
ppm
from
Week
8
through
the
end
of
the
study.
Discolored
feces
were
observed
at
6000/
4500
ppm.
Body
weights
were
decreased
(
p<=
0.05)
in
males
during
Weeks
6­
9
(­
9
to
­
20%)
and
in
females
during
Weeks
4­
9
(­
13
to
­
27%).
Cumulative
body
weight
gain
was
decreased
(­
56
to
­
109%;
p

0.05)
in
males
in
Weeks
6
and
7
and
in
females
from
Weeks
4
to
8.
Increased
incidences
of
histopathological
lesions
of
the
tongue
were
observed
in
both
sexes,
including
slight
to
marked
serous
exudate,
minimal
to
severe
ulceration,
and
minimal
to
slight
chronic
active
inflammation.
These
lesions
of
the
tongue
appear
to
follow
a
consistent
progression
and
likely
contribute
to
decreased
food
consumption
and
decreased
body
weights.
The
only
finding
observed
at
300
ppm
was
excessive
salivation
in
females.
The
LOAEL
is
3000
ppm
(
equivalent
to
106/
108
mg/
kg/
day
[
M/
F])
based
on
clinical
signs
(
tongue
erosions/
ulcerations,
few
feces,
swollen
mouth,
excessive
salivation,
and
thickened
skin
at
the
commissure
of
the
mouth),
atrophy
of
the
dorsal
epithelium
of
the
tongue,
and
loss
of
epithelial
papillae
of
the
dorsal
surface
of
the
tongue
in
both
sexes.
The
NOAEL
is
300
ppm
(
equivalent
to
11/
11
mg/
kg/
day
[
M/
F]).
This
study
is
classified
as
acceptable/
guideline
and
satisfies
the
guideline
requirements
(
OPPTS
870.3150;
OECD
409)
for
a
subchronic
oral
toxicity
study
in
the
dog.
Page
37
of
48
A­
2.4
Chronic
Oral
Toxicity
­
Dog
(
870.4100b)
In
a
chronic
oral
toxicity
study
(
MRID
#
46185901),
kasugamycin
hydrochloride
hydrate
(
72.8%
ai;
dose
levels
adjusted
for
purity;
Lot
#
KMB­
0027)
was
administered
to
4
beagle
dogs/
sex/
group
in
the
diet
at
dose
levels
of
0,
300,
1000,
or
3000
ppm
(
equivalent
to
10.5/
9.4,
30.5/
33.4,
99.6/
103.6
mg/
kg/
day
[
M/
F],
respectively)
for
52
weeks.
No
adverse
treatment­
related
effects
were
observed
on
mortality,
clinical
signs,
body
weight,
body
weight
gain,
food
consumption,
food
efficiency,
ophthalmoscopy,
hematology,
organ
weights,
or
gross
or
microscopic
pathology.
Gross
and
histopathological
tongue
lesions
that
were
observed
in
both
sexes
in
a
concurrently
submitted
13­
week
feed
study
(
MRID
#
46030302)
at
doses
of
3000
and
6000/
4500
ppm
were
not
observed
in
this
study
at
3000
ppm.
The
LOAEL
was
not
observed.
The
NOAEL
is
3000
ppm
(
equivalent
to
99.6/
103.6
mg/
kg/
day
[
M/
F]),
the
highest
dose
tested
in
the
study.
Although
the
investigators
did
not
test
to
the
limit
dose
and
a
LOAEL
was
not
observed,
the
doses
selected
were
appropriate
based
on
results
from
a
13­
week
subchronic
study
in
dogs
in
which
tongue
lesions
were
observed
at
3000
ppm
and
6000/
4500
ppm
that
affected
food
consumption
and
body
weight.
This
study
is
classified
as
acceptable/
guideline
and
satisfies
the
guideline
requirements
(
OPPTS
870.4100b;
OECD
452)
for
a
chronic
oral
toxicity
study
in
the
dog.

A­
2.5
Carcinogenicity
­
Mouse
(
870.4200b)
In
a
carcinogenicity
study
(
MRID
#
46030304),
kasugamycin
hydrochloride
hydrate
(
64.5­
64.9%
ai,
dose
levels
adjusted
for
purity,
Lot
#
KP­
821)
was
administered
to
72
CD­
1
mice/
sex/
group
in
the
diet
at
dose
levels
of
0,
50,
300,
or
1500
ppm
(
equivalent
to
0/
0,
5.93/
7.25,
34.94/
42.49,
and
186.3/
215.2
mg/
kg/
day
[
M/
F],
respectively)
for
up
to
78
weeks,
with
an
interim
sacrifice
of
twenty
mice/
sex/
group
at
Week
52.
No
adverse
treatment­
related
effects
were
observed
on
mortality,
body
weight,
body
weight
gain,
food
consumption,
food
efficiency,
hematology,
organ
weights,
gross
pathology,
or
microscopic
(
non­
neoplastic
or
neoplastic)
findings.
The
LOAEL
was
not
observed.
The
NOAEL
is
1500
ppm
(
equivalent
to
186.3/
215.2
mg/
kg/
day
[
M/
F]),
the
highest
dose
in
the
study.
Although
the
investigators
did
not
test
to
the
limit
dose
and
a
LOAEL
was
not
observed,
the
doses
selected
were
appropriate
based
on
results
from
a
13­
week
subchronic
oral
toxicity
study
in
which
mice
were
dosed
at
0,
300,
1000,
3000,
or
10,000
ppm
(
MRID
#
45910019).
In
the
subchronic
study,
increased
mortality
and
histopathological
changes
in
the
kidney
(
basophilia/
hyperplasia
in
the
pars
recta)
and
anus
(
chronic
active
inflammation,
ulceration,
and
perianal
reddening)
were
observed
at
3000
ppm.
Under
the
conditions
of
this
study,
the
carcinogenic
potential
of
kasugamycin
hydrochloride
hydrate
is
negative.
A
sufficient
maximum
tolerated
dose
(
MTD)
was
not
achieved
in
this
study.
However,
kasugamycin
hydrochloride
hydrate
was
negative
for
mutagenicity
in
concurrently
reviewed
studies,
including
the
V79/
HGPRT
forward
mutation
assay
(
MRID
#
45910026),
a
cytogenetic
assay
for
chromosome
aberrations
in
CHO
cells
(
MRID
#
45910025),
a
bacterial
gene
mutation
and
DNA
damage
assay
(
MRID
#
45910028),
an
in
vivo
micronucleus
assay
in
mice
(
MRID
#
46030305),
and
a
study
for
unscheduled
DNA
synthesis
(
MRID
#
45910027).
Additionally,
the
carcinogenic
potential
of
kasugamycin
hydrochloride
hydrate
was
negative
in
a
concurrently
reviewed
24­
month
oral
chronic
toxicity
and
carcinogenicity
study
in
Page
38
of
48
rats
(
MRID
#
45910024).
This
study
is
classified
as
acceptable/
guideline
and
satisfies
the
guideline
requirements
(
OPPTS
870.4100;
OECD
452)
for
a
chronic
oral
toxicity
study
in
the
mouse.

A­
2.6
Mutagenicity
­
Bacterial
Gene
Mutation
(
870.5100)
and
Bacterial
DNA
Damage
(
870.5500)
In
this
study
(
MRID
#
45910028),
three
different
bacterial
assays
were
performed.
In
a
DNA
damage
assay,
Bacillus
subtilis
strains
H17
and
M45
were
exposed
overnight
to
kasugamycin
HCl
(
purity:
80%
ai;
Lot/
Batch
#:
not
reported)
in
distilled
water
at
concentrations
of
0,
20,
100,
200,
1000,
or
2000

g/
disk.
The
diffusion
test
method
was
used.
In
an
in
vitro
gene
mutation
assay,
S.
typhimurium
strains
TA98,
TA100,
TA1535,
TA1537,
and
TA1538
and
E.
coli
strain
WP2hcr
were
exposed
to
kasugamycin
HCl
in
distilled
water
at
concentrations
of
0,
5,
10,
50,
100,
or
200

g/
plate
in
the
presence
and
absence
of
S9­
activation.
Additionally,
S.
typhimurium
strain
G46
(
his­)
was
similarly
tested
at
concentrations
of
0,
5,
10,
50,
100,
or
500

g/
plate
in
the
absence
of
S9­
activation
only.
Strain
specific
positive
controls
were
used.
In
a
non­
guideline
host­
mediated
gene
mutation
assay,
5­
6
male
ICR
mice/
dose
received
two
equal
doses
(
gavage,
20
mL/
kg)
of
kasugamycin
HCl
in
distilled
water
over
a
24
hour
period
at
doses
of
500
or
2000
mg/
kg
(
total
dose
received
was
1000
or
4000
mg/
kg).
Immediately
after
the
second
dosing,
a
2
mL
suspension
of
S.
typhimurium
strain
G46
(
his­)
was
inoculated
via
i.
p.
injection.
Three
hours
after
the
final
treatment,
the
peritoneal
fluid
was
removed
and
used
in
an
Ames
reverse
mutation
assay.
In
the
DNA
damage
assay,
kasugamycin
HCl
was
tested
up
to
2000

g/
disk.
No
treatment­
related
increases
in
the
difference
between
zones
of
inhibition
were
observed
in
any
treatment
group
compared
to
the
concurrent
controls.
The
positive
controls
induced
the
appropriate
response.
There
was
no
evidence
that
DNA
damage
was
induced.
The
study
is
classified
as
acceptable/
guideline
and
satisfies
the
guideline
requirements
(
OPPTS
870.5500)
for
other
genotoxic
mutagenicity
(
bacterial
DNA
damage)
data.
In
the
gene
reverse
mutation
assay,
kasugamycin
HCl
was
tested
up
to
500

g/
plate
in
S.
typhimurium
strain
G46
(
his­)
in
the
absence
of
S9,
and
up
to
200

g/
plate
in
all
other
strains
(+/­
S9).
It
was
not
reported
if
any
evidence
of
cytotoxicity
(
thinning
of
the
background
lawn
or
reduction
in
number
of
revertants)
was
observed.
No
treatment­
related
increases
in
the
mean
number
of
revertants/
plate
were
observed
in
any
bacterial
strain
at
any
dose
level
in
the
presence
or
absence
of
S9­
activation
compared
to
solvent
controls.
The
positive
controls
induced
the
appropriate
response.
There
was
no
evidence
of
induced
mutant
colonies
over
background
in
the
presence
or
absence
of
S9.
Because
the
test
material
was
not
evaluated
at
the
limit
dose
(
5000

g/
plate);
there
was
no
indication
of
test
material
induced
cytotoxicity;
and
there
was
no
defined
limit
of
solubility,
it
could
not
be
determined
if
a
sufficiently
high
dose
was
used.
Therefore,
the
study
is
classified
as
unacceptable/
upgradable
and
does
not
satisfy
the
guideline
requirements
(
OPPTS
870.5100;
OECD
471)
for
in
vitro
mutagenicity
(
bacterial
reverse
gene
mutation).
The
study
may
be
upgraded
to
acceptable
pending
submission
of
data
that
indicate
that
the
high
dose
was
sufficient
(
e.
g..
test
material
solubility
data
or
cytotoxicity
data).
In
the
host­
mediated
assay,
kasugamycin
HCl
was
tested
up
to
4000
mg/
kg
(
total
dose).
No
significant
increases
in
the
mean
mutation
frequency
were
observed
at
either
dose
compared
to
controls.
The
positive
control
induced
the
appropriate
response.
There
was
no
evidence
of
induced
mutant
colonies
over
background.
The
study
is
classified
as
acceptable/
nonguideline
Page
39
of
48
A­
2.7
Mutagenicity
­
Mammalian
Gene
Mutation
(
870.5300)
In
three
independent
mammalian
cell
gene
mutation
assays
at
the
HGPRT
locus
(
MRID
#
45910026)
in
the
presence
of
S9­
activation
and
two
independent
studies
in
the
absence
of
S9­
activation,
Chinese
hamster
V79
lung
fibroblasts
cultured
in
vitro
were
exposed
to
kasugamycin
hydrochloride
hydrate
(
67.1%
ai;
Lot
#
KP­
570)
in
Eagle's
minimal
essential
medium
at
concentrations
of
0.5,
1,
2,
4,
6,
8,
or
10
mg/
mL
for
4
hours.
Kasugamycin
hydrochloride
hydrate
was
tested
up
to
cytotoxic
concentrations
(
10
mg/
mL
[
Trial
1,
­
S9
and
Trial
2,
+
S9]
and
>=
8
mg/
mL
[
Trial
1,
+
S9]).
No
treatment­
related
increases
in
mutant
frequency
were
observed
in
any
trial
in
the
presence
or
absence
of
S9.
The
positive
controls
induced
the
appropriate
response.
There
was
no
evidence
that
kasugamycin
hydrochloride
hydrate
induced
mutant
colonies
over
background
in
the
presence
or
absence
of
S9­
activation.
This
study
is
classified
as
acceptable/
guideline
and
satisfies
the
guideline
requirements
(
OPPTS
870.5300,
OECD
476)
for
in
vitro
mutagenicity
(
mammalian
forward
gene
mutation)
data.

A­
2.8
Mutagenicity
­
Structural
Chromosomal
Aberrations
(
870.5375)
In
a
mammalian
cell
cytogenetics
assay
(
chromosome
aberration;
MRID
#
45910025),
Chinese
hamster
ovary
cell
cultures
were
exposed
to
kasugamycin
hydrochloride
hydrate
(
67.1%
ai;
Lot
#
KP­
570)
in
McCoy's
5a
medium
at
concentrations
of
1,
2,
3,
4,
or
5
mg/
mL
for
2
hours
in
the
presence
of
S9
or
for
7.25
hours
in
the
absence
of
S9.
Cells
were
harvested
at
8
or
2.5
hours
after
conclusion
of
treatment
in
the
presence
or
absence
of
S9,
respectively.
Kasugamycin
hydrochloride
was
tested
up
to
5
mg/
mL
(+/­
S9).
Although
only
evidence
of
slight
cytotoxicity
was
observed
at
5
mg/
mL
(+/­
S9)
in
the
cytogenetic
assay,
the
results
of
the
preliminary
study
indicate
that
the
dose
levels
selected
for
the
main
study
were
acceptable.
There
were
no
significant
increases
in
the
percentage
of
cells
with
aberrations
(
excluding
gaps)
at
any
dose
in
the
presence
or
absence
of
S9.
The
most
frequently
observed
types
of
aberration
(
excluding
gaps)
were
acentric
fragments
and
dicentrics.
The
positive
controls
induced
the
appropriate
response.
There
was
no
evidence
that
kasugamycin
hydrochloride
induced
a
clastogenic
effect
in
the
presence
or
absence
of
S9­
activation.
However,
the
current
guidelines
recommend
a
harvest
time
of
1.5
cell
cycles
after
treatment.
The
cell
cycle
for
CHO
cells
is
approximately
12­
14
hours.
The
cells
were
harvested
at
only
10
hours
after
treatment,
and
there
was
evidence
of
a
slight
cell
cycle
delay
at
5
mg/
mL
in
the
absence
of
S9.
Therefore,
the
reviewers
conclude
that
the
time
from
treatment
to
cell
harvest
was
insufficient.
This
study
is
classified
as
unacceptable/
not
upgradable
and
does
not
satisfy
the
guideline
requirement
(
OPPTS
870.5375,
OECD
473)
for
in
vitro
mutagenicity
(
chromosome
aberration)
data.

A­
2.9
Mutagenicity
­
Mammalian
Erythrocyte
Microcnucleus
(
870.5395)
In
a
bone
marrow
micronucleus
assay
(
MRID
#
46030305),
5
non­
fasted
CD­
1
mice/
sex/
dose
were
treated
once
via
gavage
(
15
mL/
kg)
with
kasugamycin
hydrochloride
hydrate
(
67.1%
ai;
Lot
#
KP­
570)
in
distilled
water
at
doses
of
0,
200,
1000,
or
5000
mg/
kg
bw,
and
bone
marrow
cells
were
harvested
at
24
hours
post­
dosing.
Additionally,
5
mice/
sex/
sacrifice
time
were
similarly
dosed
at
0
or
5000
mg/
kg,
and
bone
marrow
cells
were
harvested
at
48
or
72
hours
post­
dosing.
Chlorambucil
(
30
mg/
kg)
served
as
the
positive
control.
Kasugamycin
hydrochloride
hydrate
was
tested
up
to
5000
mg/
kg;
however,
no
evidence
of
bone
marrow
Page
40
of
48
toxicity
(
decreased
PCE:
NCE)
was
observed
at
any
dose
at
24,
48,
or
72
hours
post­
dosing.
Clinical
signs
of
toxicity
were
limited
to
brown
anal
staining
and
diarrhea
at
5000
mg/
kg
on
the
day
of
dosing.
No
significant
increase
in
micronucleated
polychromatic
erythrocytes
(
MPCEs)
was
observed
at
24,
48,
or
72
hours
post­
dosing.
The
positive
control
induced
the
appropriate
response.
There
was
no
significant
increase
in
the
frequency
of
micronucleated
polychromatic
erythrocytes
in
bone
marrow
compared
to
controls.
The
study
is
classified
as
acceptable/
guideline
and
satisfies
the
guideline
requirement
(
OPPTS
870.5395;
OECD
474)
for
in
vivo
cytogenetic
mutagenicity
data.

A­
2.10
Mutagenicity
­
Unscheduled
DNA
Synthesis
in
Mammalian
Cells
in
Culture
(
870.5550)
In
three
independent
trials
of
an
unscheduled
DNA
synthesis
assay
(
MRID
#
45910027),
HeLa
S3
cell
cultures
were
exposed
to
kasugamycin
hydrochloride
hydrate
(
67.1%
ai;
Lot
No.:
KP­
570)
in
arginine­
free
Eagle's
Minimal
Essential
Medium
for
3
hours
at
concentrations
of
0.156,
0.313,
0.625,
1.25,
or
2.50
mg/
mL
(+/­
S9;
Trial
1)
and
0.313,
0.625,
1.25,
2.50,
5.00,
or
10.0
mg/
mL
(+/­
S9;
Trial
2
and
+
S9;
Trial
3).
Kasugamycin
hydrochloride
hydrate
was
tested
up
to
cytotoxic
concentrations
in
the
presence
of
S9
(
10
mg/
mL,
Trial
3).
Although
no
evidence
of
cytotoxicity
(
decreased
amount
of
DNA
recovered)
was
observed
at
up
to
10
mg/
mL
in
the
absence
of
S9,
the
results
of
the
preliminary
cytotoxicity
test
indicate
that
10
mg/
mL
was
the
appropriate
high­
dose
for
the
UDS
assay.
There
were
no
reproducible
increases
(>
50%
higher
than
the
controls)
in
the
incorporation
of
[
3H]
thymidine
in
the
presence
or
absence
of
S9
at
any
dose.
The
positive
controls
induced
the
appropriate
response
in
all
trials.
There
was
no
evidence
that
unscheduled
DNA
synthesis,
as
determined
by
liquid
scintillation
counting
procedures,
was
induced.
The
study
is
classified
as
acceptable/
guideline
and
satisfies
the
guideline
requirement
(
OPPTS
870.5550;
OECD
482)
for
other
genotoxic
mutagenicity
data.

A­
2.11
Metabolism
­
Rat
(
870.7485)
In
a
rat
metabolism
study
(
MRID
#
46030306),
[
14C]­
kasugamycin
hydrochloride
hydrate
in
sterile
deionized
water
was
administered
to
Fischer
344
(
F344­
NHLa
CFV)
rats
by
gavage.
In
the
main
ADME
study,
5
rats/
sex/
dose
were
given
[
14C]­
kasugamycin
hydrochloride
hydrate
(
Batch
#
CP­
1916;
radiochemical
purity
>
98%)
either
as
a
single
oral
dose
of
100
or
1000
mg/
kg,
or
as
a
repeated
oral
dose
of
100
or
1000
mg/
kg
(
14
days
unlabeled
+
1
day
radiolabeled).
Pharmacokinetic,
tissue
distribution
time
course,
and
biliary
excretion
studies
were
also
performed
on
males
and
females
at
both
dose
levels.
Metabolites
were
identified
and
quantified
in
the
urine
and
feces
from
the
main
ADME
study
and
in
plasma,
kidney,
and
liver
from
the
tissue
distribution
time
course
study.
In
the
main
ADME
study,
the
total
recovery
of
the
radioactive
dose
ranged
from
90.7­
96.8%
dose
at
168
hours
post­
dose,
with
the
majority
of
the
dose
recovered
within
48
hours
in
the
feces
(
81.9­
93.9%)
and
urine
(
1.26­
3.07%).
Fecal
excretion
accounted
for
87.7­
94.5%
in
all
groups;
urinary
excretion
accounted
for
1.35­
3.26%;
and
the
cage
rinse,
wash,
and
wipe
accounted
for
<=
1.14%.
The
carcass/
tissues
retained
<=
0.13%
dose,
and
results
from
a
previous
study
indicated
a
minimum
amount
of
14CO
2
(<
0.2%
of
total
radioactivity)
was
detected
in
exhaled
air.
For
the
bile­
duct
cannulated
rats,
<
0.01%
of
the
dose
was
recovered
in
the
bile.
For
both
sexes,
concentrations
of
radioactivity
in
plasma
peaked
within
1
hour
of
dosing
Page
41
of
48
for
both
the
100
mg/
kg
dose
group
(
1.47­
2.17

g/
g)
and
the
1000
mg/
kg
dose
group
(
5.23­
6.40

g/
g)
and
declined
to
below
the
limit
of
quantitation
by
24
hours
post­
dose.
A
terminal
half
life
of
radioactivity
of
1.17­
1.55
hours
was
observed
in
plasma.
The
total
dose
absorbed
as
indicated
by
the
area
under
the
curve
(
AUC)
was
proportional
to
dose
in
both
sexes
at
both
100
mg/
kg
(
2.88­
3.63

g*
hour/
g)
and
1000
mg/
kg
(
15.3­
18.7

g*
hour/
g).
The
time
course
of
tissue
distribution
showed
that
the
concentration
of
radioactivity
was
higher
in
the
kidneys,
urinary
bladder,
and
lymph
nodes
than
in
blood
in
both
sexes
at
all
time
points
in
both
the
low
(
0.582­
30.3

g
Eq/
g
vs
<=
1.80

g
Eq/
g
in
blood)
and
high
(
6.43­
147.0

g
Eq/
g
vs
<=
8.32

g
Eq/
g
in
blood)
dose
groups.
In
the
pancreas,
increases
over
blood
levels
were
also
observed
in
the
low­
and
high­
dose
females
at
all
time
points
and
in
the
low­
and
high­
dose
males
at
selected
time
points.
Concentrations
of
radioactivity
were
also
approximately
twice
as
high
in
plasma
compared
to
whole
blood.
All
other
tissues
(
excluding
G.
I.
tract)
had
concentrations
of
radioactivity
that
were
comparable
to
or
below
the
concentrations
in
the
blood
throughout
the
time
course.
By
168
hours
post­
dose,
concentrations
of
radioactivity
were
nondetectable
in
all
but
a
few
tissues,
with
the
highest
concentrations
being
detected
in
kidneys
(
1.89­
24.7

g
Eq/
g)
in
both
sexes
and
in
all
groups.
Radioactivity
was
also
detected
in
the
intestinal
tract
(
0.92­
1.11

g
Eq/
g)
and
stomach
(
1.14­
1.21

g
Eq/
g)
of
both
sexes
from
the
1000
mg/
kg
single
dose
group,
and
in
the
intestinal
tract
(
0.48

g
Eq/
g)
of
females
from
the
1000
mg/
kg
repeated
dose
females.
The
metabolism
of
kasugamycin
hydrochloride
hydrate
in
rats
was
not
affected
by
sex,
dose
level,
or
duration
of
dosing.
Parent
was
the
major
component
identified
in
the
urine,
feces,
liver,
kidney
and
plasma,
and
minor
amounts
(<
1%
dose)
of
the
metabolite
kasuganobiosamine
were
identified
in
urine,
liver,
kidney
and
plasma.
The
presence
of
kasugamycinic
acid
was
also
postulated
in
liver
extracts.
Parent
compound
was
identified
by
TLC
and
HPLC;
metabolites
were
identified
by
mass
spectroscopy.
In
the
main
ADME
study
(
Groups
A­
D),
parent
was
the
predominant
compound
identified
in
both
feces
(
79.9­
89.8%
dose)
and
urine
(
1.3­
2.9%
dose)
in
all
groups,
totaling
81.6­
92.7%
dose.
In
the
urine
from
all
groups,
kasuganobiosamine
was
identified
by
mass
spectroscopy,
but
was
not
quantified
as
the
metabolite
could
not
be
resolved
from
the
parent
by
HPLC.
Kasuganobiosamine
was
not
detected
in
the
feces.
Parent
and
kasuganobiosamine
were
also
detected
in
the
kidney
and
liver
extracts
from
animals
sacrificed
1­
6
hours
post­
dose,
and
kasugamycinic
acid
was
also
putatively
detected
in
the
liver.
Radioactivity
in
the
kidney
and
liver
accounted
for
<
0.01­
0.18%
of
the
dose
in
these
samples
and

0.03%
of
the
dose
in
animals
sacrificed
at
168
hours
post­
dose.
Parent
and
kasuganobiosamine
were
also
reportedly
detected
in
plasma.
Regardless
of
sex,
dose
level
(
100
or
1000
mg/
kg),
and
duration
of
dosing,
the
absorption
and
metabolism
of
[
14C]­
kasugamycin
hydrochloride
hydrate
in
rats
was
limited
(<
5%
dose).
This
metabolism
study
in
the
rat
is
classified
acceptable/
guideline
and
satisfies
the
guideline
requirement
for
a
Tier
1
metabolism
study
[
OPPTS
870.7485,
OPP
85­
1]
in
rats.

A­
3.0
METABOLISM
CONSIDERATIONS
A­
3.1
Team
Proposal
The
only
metabolite
which
would
possibly
be
considered
for
inclusion
in
the
ROC
(
in
addition
to
the
parent)
was
kasugamycinic
acid,
which
was
present
at
less
than
10%
in
all
foliage
Page
42
of
48
samples,
and
only
occurred
at
concentrations
of
10%
or
more
in
fruit
samples
from
the
21­
and
28­
day
PHIs.
Because
the
petitioner
is
proposing
a
PHI
of
1­
day
on
imported
fruiting
vegetables,
kasugamycinic
acid
is
unlikely
to
occur
as
other
than
a
minor
constituent
in
imported
fruiting
vegetable
commodities,
and
was
therefore
not
included
in
the
ROC
for
either
tolerance
expression
or
risk
assessment
purposes.
Also,
as
residues
of
kasugamycin
in
all
samples
from
the
crop
field
trials
(
at
1X)
were
either
non­
detectable
or
less
than
the
LOQ,
the
increase
in
risk
from
inclusion
of
this
relatively
minor
metabolite
was
expected
to
be
negligable.

A­
3.2
Nature
of
the
Residue
Studies
in
Plants
Executive
Summary
of
Tomato
Metabolism
Study
Arvesta
Corporation,
on
behalf
of
Hokko
Chemical
Industry
Company,
Limited
(
Japan),
has
submitted
a
study
investigating
the
metabolism
of
[
14C]­
kasugamycin
in
tomatoes.
The
radiolabeled
test
substance
was
applied
as
a
single
foliar
broadcast
spray
to
tomato
plants
at
approximately
0.17
pound
active
ingredient
per
acre
(
lb
ai/
A),
slightly
more
than
3X
the
proposed
maximum
seasonal
application
rate.
Mature
tomatoes
and
foliage
were
harvested
2
hours
and
1,
7,
14,
21,
and
28
days
following
application.
Residues
were
higher
in
the
tomato
foliage
than
in
the
fruit.
TRR,
determined
by
summing
extractable
and
nonextractable
radioactivity,
were
0.011,
0.008,
0.027,
0.072,
0.098,
and
0.084
ppm
in
tomatoes
harvested
2
hours,
1
day,
7
days,
14
days,
21
days,
and
28
days
after
treatment,
respectively.
TRR
in
foliage
from
the
respective
pre­
harvest
intervals
(
PHIs)
were
2.86,
4.73,
1.77,
2.92,
2.57,
and
4.29
ppm.
Extraction
with
water
removed
91­
94%
of
the
TRR
from
fruit
and
80­
86%
TRR
from
foliage.
Extraction
with
methanol
released
an
additional
2­
3%
TRR
from
fruit
with
14­
through
28­
day
PHIs
and
4­
8%
TRR
from
the
corresponding
foliage.
Nonextractable
residues
accounted
for
a
maximum
of
9%
TRR
in
fruit
and
no
more
than
12%
TRR
in
foliage.
Residues
were
characterized
or
identified
primarily
by
HPLC
analysis,
with
confirmatory
analysis
by
TLC.
These
methods
successfully
identified
the
predominant
residues
in
tomatoes
and
foliage.
Approximately
90%
and
94%
of
the
TRR
were
identified
in
tomatoes
harvested
2
hours
and
1
day
after
treatment,
respectively,
and
55­
69%
TRR
were
identified
in
tomatoes
harvested
7­
28
days
after
harvest.
In
foliage,
roughly
86%
and
80%
TRR
were
identified
in
samples
harvested
2
hours
and
1
day
after
treatment,
respectively,
and
60­
72%
TRR
were
identified
in
foliage
from
the
remaining
sampling
intervals.
In
general
the
metabolite
profile
was
similar
in
tomato
fruit
and
foliage.
Parent
compound
(
kasugamycin
per
se)
was
the
major
identified
component
in
all
samples
from
all
harvest
intervals,
accounting
for
90.4­
93.9%
TRR
in
tomatoes
from
the
2­
hour
and
1­
day
PHIs,
for
69.2%
TRR
in
tomatoes
from
the
7­
day
PHI,
and
for
54.8­
59.5%
TRR
in
tomatoes
from
the
remaining
sampling
intervals.
In
foliage,
kasugamycin
accounted
for
75.0­
84.0%
TRR
in
samples
from
the
2­
hour
and
1­
day
PHIs,
for
69.5%
TRR
in
samples
from
the
7­
day
PHI,
and
for
52.5­
57.2%
TRR
in
foliage
from
the
remaining
sampling
intervals.
In
tomatoes,
the
only
other
identified
metabolite
was
kasugamycinic
acid,
which
was
found
in
tomatoes
from
the
21­
and
28­
day
PHIs
at
9.5%
and
12.0%
TRR,
respectively.
Kasugamycinic
acid
was
also
identified
in
foliage
from
all
harvest
intervals
at
2.4­
7.1%
TRR.
Two
additional
metabolites
were
identified
in
foliage:
2­
N­
acetyl­
kasugamycin
was
identified
in
14­
through
28­
day­
PHI
foliage
at
0.7­
2.2%
TRR,
and
kasuganobiosamine
was
identified
in
1­
through
28­
day­
PHI
tomato
foliage
at
0.4­
1.0%
TRR.
Remaining
radioactivity
was
characterized
as
unknowns,
each
present
at
maximum
concentrations
of
19.5%
TRR
in
fruit
and
12.2%
TRR
in
foliage.
Unknowns
accounting
for
13.5­
19.5%
TRR
in
fruit
and
2.9­
12.2%
TRR
in
foliage
were
subjected
to
further
Page
43
of
48
characterization
procedures
which
confirmed
that
they
were
comprised
of
multiple
minor
components.

Based
on
the
results
of
the
tomato
metabolism
study
the
petitioner
proposes
that
the
major
metabolic
pathway
of
kasugamycin
in
tomatoes
involves
conjugation
of
the
parent
compound,
conversion
to
kasugamycinic
acid,
and
subsequent
conjugation
of
kasugamycinic
acid.
Conversion
of
kasugamycin
to
2­
N­
acetyl
kasugamycin
and
kasuganobiosamine
was
thought
to
be
a
minor
metabolic
route.
Page
44
of
48
Tabular
Summary
of
Tomato
Metabolism
Study
APPENDIX
TABLE
2
Summary
of
Characterization
&
Identification
of
Radioactive
Residues
in
Tomato
Fruit
Following
Foliar
Broadcast
Application
of
[
14C]­
Kasugamycin
at
0.17
lb
ai/
A.

Compound
2­
hour
PHI
1­
Day
PHI
7­
Day
PHI
14­
Day
PHI
21­
Day
PHI
28­
Day
PHI
TRR
=
0.011
ppm
TRR
=
0.008
ppm
TRR
=
0.027
ppm
TRR
=
0.072
ppm
TRR
=
0.098
ppm
TRR
=
0.084
ppm
%
TRR
ppm
%
TRR
ppm
%
TRR
ppm
%
TRR
ppm
%
TRR
ppm
%
TRR
ppm
Kasugamycin
90.4
0.010
93.9
0.007
69.2
0.018
54.8
0.039
59.5
0.058
57.3
0.049
Kasugamycinic
acid
­­
­­
­­
­­
­­
­­
­­
­­
9.5
0.009
12.0
0.010
Unknown
1
­­
­­
­­
­­
­­
­­
3.2
0.002
­­
­­
­­
­­

Unknown
2
­­
­­
­­
­­
13.5
0.004
19.5
0.014
15.6
0.015
14.1
0.012
Unknown
3
­­
­­
­­
­­
5.5
0.001
7.7
0.006
5.5
0.005
7.1
0.006
Unknown
4
­­
­­
­­
­­
4.9
0.001
3.4
0.002
­­
­­
­­
­­

Others
­­
­­
­­
­­
­­
­­
2.3
0.002
­­
­­
­­
­­

Methanol
extractable
­­
­­
­­
­­
­­
­­
2.8
0.002
3.1
0.003
2.7
0.002
Total
identified
90.4
0.010
93.9
0.007
69.2
0.018
54.8
0.039
69.0
0.067
69.3
0.059
Total
characterized
­­
­­
­­
­­
23.9
0.006
38.9
0.028
24.2
0.023
23.9
0.020
Total
extractable
91.0
0.010
94.1
0.007
93.7
0.025
93.1
0.067
93.7
0.092
93.5
0.079
Unextractable
(
PES)
9.0
0.001
5.9
<
0.001
6.3
0.002
6.9
0.005
6.3
0.006
6.5
0.005
Accountability*
100
100
100
100
100
100
*
Accountability
=
Total
Extractable
+
Total
Unextractable.
Accountabilities
were
100%
because
the
TRR
were
determined
by
summing
extractable
and
nonextractable
residues.
Page
45
of
48
APPENDIX
TABLE
3
Summary
of
Characterization
&
Identification
of
Radioactive
Residues
in
Tomato
Foliage
Following
Foliar
Broadcast
Application
of
[
14C]­
Kasugamycin
at
0.17
lb
ai/
A.

Compound
1
2­
hour
PHI
1­
Day
PHI
7­
Day
PHI
14­
Day
PHI
21­
Day
PHI
28­
Day
PHI
TRR
=
2.86
ppm
TRR
=
4.73
ppm
TRR
=
1.77
ppm
TRR
=
2.92
ppm
TRR
=
2.57
ppm
TRR
=
4.29
ppm
%
TRR
ppm
%
TRR
ppm
%
TRR
ppm
%
TRR
ppm
%
TRR
ppm
%
TRR
ppm
Kasugamycin
84.0
2.40
75.0
3.54
69.5
1.23
56.7
1.66
57.2
1.47
52.5
2.25
Kasugamycinic
acid
2.4
0.069
5.0
0.234
2.5
0.045
6.5
0.191
6.2
0.159
7.1
0.304
2­
N­
acetyl­
kasugamycin
­­
 
­
­
 
­
­
 
2.2
0.064
1.1
0.027
0.7
0.030
Unknown
1
1.3
0.036
2.6
0.121
2.3
0.040
2.7
0.080
2.8
0.073
4.9
0.210
Unknown
2
­­
­­
0.8
0.038
5.1
0.090
1.9
0.057
5.4
0.138
6.9
0.297
Unknown
3
­­
­­
2.9
0.138
7.7
0.137
11.3
0.330
10.2
0.263
12.2
0.522
Unknown
4
1.7
0.049
­­
­­
­­
­­
1.5
0.044
1.9
0.048
1.8
0.075
Others
­­
­­
2.0
0.096
­­
­­
4.7
0.136
3.4
0.088
1.1
0.045
Total
identified
86.4
2.47
80.0
3.78
72.0
1.28
65.4
1.91
64.5
1.66
60.3
2.58
Total
characterized
3.0
0.085
8.3
0.393
15.1
0.267
22.1
0.647
23.7
0.610
26.9
1.15
Total
extractable
89.7
2.57
88.4
4.18
87.7
1.56
88.1
2.58
88.3
2.27
87.7
3.76
Nonextractable
10.3
0.296
11.6
0.550
12.3
0.218
12.0
0.350
11.7
0.300
12.4
0.531
Accountability
2
100
100
100
100
100
100
1.
An
additional
metabolite,
kasuganobiosamine,
not
detected
by
HPLC
analysis,
was
identified
by
TLC
in
1­
through
28­
day­
PHI
tomato
foliage
at
0.4­
1.0%

TRR
(
0.008­
0.026
ppm).

2.
Accountability
=
Total
Extractable
+
Total
Unextractable.
Accountabilities
were
100%
because
the
TRR
were
determined
by
summing
extractable
and
nonextractable
residues.
Page
46
of
48
A­
4.0
ANALYTICAL
METHODOLOGY
APPENDIX
TABLE
4
Summary
of
Parameters
for
the
Proposed
Tolerance
Enforcement
Method
for
Kasugamycin
Residues
in
Plant
Matrices.

Method
Name
Morse
Laboratories
Analytical
Method
#
Meth­
146.

Applicable
Commodities
Fruiting
Vegetables
(
Crop
Group
8).

Analyte
Kasugamycin.

Extraction
Solvents
Frozen,
macerated
samples
are
combined
with
methanol/
de­
ionized
(
DI)
water
(
7:
3
vol/
vol)
and
adjusted
to
pH
4
with
6
N
HCl,
then
extracted
via
homogenization
and
vacuum
filtered.
The
extraction
is
repeated
at
pH
4
using
1
N
HCl
for
acidification.
Filtrates
are
combined,
concentrated,
and
cooled
to
precipitate
impurities,
then
vacuum
filtered
through
Celite
(
diatomaceous
earth)
and
reconstituted
with
DI
water.

Clean­
Up
Steps
Two
separate
ion
exchange
columns
are
used
for
clean­
up.
The
purified
extract
is
applied
to
a
chromatographic
column
containing
Dowex
50WX8
resin
(
100­
200,
H
form);
residues
of
kasugamycin
are
eluted
with
0.5%
ammonia
solution.
The
eluate
is
concentrated
and
adjusted
to
pH
6
to
7
with
0.2
N
HCl,
then
reconstituted
with
DI
water,
and
applied
to
a
second
chromatographic
column
containing
Amberlite
CG­
50
resin
(
NH4
+/
H+,
7:
3
vol/
vol);
residues
of
kasugamycin
are
eluted
with
1.0%
ammonia
solution.
The
eluate
is
concentrated
to
dryness,
then
redissolved
in
0.04%
phosphoric
acid
with
sonication
to
help
dissolve
the
residues.
The
petitioner
notes
that
the
pH
of
the
purified
extract
must
be
less
than
7.

Determinative
Step
Analysis
is
by
ion­
pairing
HPLC
utilizing
a
reverse­
phase
C­
18
column
with
UV
detection
(
230
nm)
for
quantitation
and
a
gradient
mobile
phase
of
acetonitrile:
water
(
18.5:
81.5
vol/
vol)
at
pH
3.5,
and
acetonitrile:
water
(
50:
50
vol/
vol)
at
pH
3.5,
each
containing
0.1%
sodium
decane
sulfonate,
by
weight,
as
the
ion­
pairing
agent.

LOQ
(
ppm)
0.040
LOD
(
ppm)
0.013
A­
5.0
SUMMARY
OF
MAGNITUDE
OF
THE
RESIDUE
STUDIES
FOR
FRUITING
VEGETABLES
(
CROP
GROUP
8)
Crop
field
trials
are
conducted
to
determine
the
maximum
residue
which
may
be
expected
in/
on
a
raw
agricultural
commodity
as
a
result
of
the
legal
use
of
the
pesticide.
The
trials
must
Page
47
of
48
reflect
label
directions
which
would
be
expected
to
result
in
the
maximum
residue
levels;
ergo,
the
trials
should
employ
maximum
label
rates,
maximum
number
of
applications,
minimum
retreatment
interval(
s),
and
minimum
pre­
harvest
interval.
Data
generated
in
the
US
or
countries
other
than
those
where
the
petitioner
has
existing
or
proposed
uses
may
be
substituted
for
up
to
half
of
the
required
number
of
foreign
trials,
but
a
minimum
of
three
trials
must
be
from
the
countries
in
which
the
pesticide
is
marketed.
The
petitioner
should
demonstrate
that
crop
cultural
practices,
climatological
conditions,
and
use
patterns
are
substantially
similar
between
the
subject
foreign
regions
and
regions
represented
by
the
US
(
or
other)
data.
The
burden
of
proof
is
on
the
petitioner.
In
the
case
of
tolerances
to
cover
treated
commodities
imported
from
Canada
or
Mexico
only,
it
may
be
acceptable
for
more
than
50%
of
the
trials
to
be
conducted
in
the
United
States.
As
part
of
the
harmonization
process
under
the
NAFTA,
the
crop
field
trial
regions
in
the
US
guidelines
have
been
extended
into
Canada
and
efforts
are
in
progress
to
do
the
same
into
Mexico.
This
would
allow
trials
in
the
US
to
support
registration
and
tolerances
in
Canada
and
Mexico
or
vice
versa.
As
a
result,
among
these
three
countries,
for
certain
crops
most
or
all
the
field
trials
could
be
in
a
different
country
than
the
one
in
which
the
tolerance
is
to
be
established.
For
example,
if
a
tolerance
is
desired
to
cover
the
export
of
cranberries
from
Canada
to
the
US,
most
of
the
trials
could
be
conducted
in
the
northern
regions
of
the
US
even
though
the
pesticide
is
to
be
registered
in
Canada.
Similarly,
for
certain
crops
being
imported
from
Mexico,
many
of
the
trials
could
be
done
in
the
southwestern
US.
In
the
future,
if
other
countries
develop
zone
maps
employing
similar
concepts
as
were
used
for
the
NAFTA
countries,
and
the
regions
and
cultural
practices
are
demonstrated
to
be
substantially
similar
to
US
regions,
then
the
Agency
may
consider
substitution
of
US
data
for
those
countries
as
well.

APPENDIX
TABLE
5
Summary
of
Residue
Data
from
Pepper
&
Tomato
Field
Trials
with
Kasugamycin.

Crop
[
Matrix]
Total
Application
Rate
(
lb
ai/
A)
[
Exaggerated
Rate]
PHI
(
Days)
Kasugamycin
Residue
Levels
(
ppm)
1
n
Min.
Max.
HAFT
2
Mean
Std.
Dev.

Pepper
[
Fruit]
0.053
­
0.054
1
14
<
0.040
<
0.040
<
0.040
0.040
0
Tomato
[
Fruit]
0.052
­
0.054
1
16
<
0.040
<
0.040
<
0.040
0.040
0
0.270
[
5X]
1
2
0.044
0.056
0.050
0.050
0.008
1.
For
reporting
minimum
and
maximum
values
(
and
calculation
of
the
median,
mean,
and
standard
deviation),
the
LOQ
value
(
0.040
ppm;
also
the
LLMV)
was
used
for
residues
reported
as
ND
or
less
than
LOQ.
2.
HAFT
=
Highest
Average
Field
Trial.

A­
6.0
INTERNATIONAL
CONSIDERATIONS
Page
48
of
48
INTERNATIONAL
RESIDUE
LIMIT
STATUS
Chemical
Name:
3­
O­[
2­
amino­
4­[(
carboxyiminomethyl)
amino]­
2,3,4,6­
tetradeoxy­
 ­
D­
arabinohexopyranosyl
D­
chiro­
inositol
Common
Name:
Kasugamycin
X
Proposed
tolerances

Reevaluated
tolerance

Other
Date:
10/
20/
2004
Codex
Status
(
Maximum
Residue
Limits)
US
Tolerances
X
No
Codex
proposal
step
6
or
above

No
Codex
proposal
step
6
or
above
for
the
crops
requested
Petition
Number:
3F6579
DP
Barcode:
D301735
Other
Identifier:
PC
Code
230001
Residue
definition
(
step
8/
CXL):
N/
A
Reviewer/
Branch:
William
T.
Drew/
RAB2
Residue
definition:
Kasugamycin
Crop(
s)
MRL
(
mg/
kg)
Crops
Tolerance
(
ppm)

Fruiting
Vegetables,
Crop
Group
8
(
proposed)
0.04
Limits
for
Canada
Limits
for
Mexico
X
No
Limits

No
Limits
for
the
crops
requested
X
No
Limits

No
Limits
for
the
crops
requested
Residue
definition:
N/
A
Residue
definition:
N/
A
Crop(
s)
MRL
(
mg/
kg)
Crop(
s)
MRL
(
mg/
kg)

NOTE:
Per
Stephen
Funk,
10/
20/
2004.