Document ID: EPA-HQ-OPP-2002-0233-0002
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2002-09-13T04:00Z

Product
Monograph
Pseudozyma
flocculosa
strain
PF­
A22
UL
(PC
Code
119196)

Pseudozyma
flocculosa
strain
PF­
A22
UL
(TGAI)
SPORODEX
L
(EP)

The
active
ingredient
Pseudozyma
flocculosa
strain
PF­
A22
UL
and
associated
end­
use
product
SPORODEX
L,
for
the
control
of
powdery
mildew
on
roses
and
cucumbers,
are
proposed
for
temporary
registration
under
Section
17
of
the
Pest
Control
Products
Regulations
(Canada)
and
a
conditional
registration
under
Section
3(
c)(
7)(
C)
of
the
Federal
Insecticide
Fungicide
and
Rodenticide
Act
(United
States).

This
Product
Monograph
provides
a
summary
of
data
reviewed
and
the
rationale
for
the
proposed
Section
17
(Canada)
and
Section
3(
c)(
7)(
C)
(U.
S.)
registration
of
these
products.

September
2002
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
Foreword
The
submissions
for
the
registration
of
the
technical
grade
active
ingredient,
Pseudozyma
flocculosa
strain
PF­
A22
UL,
and
its
end­
use
product,
SPORODEX
L,
manufactured
by
Plant
Products
Co.,
have
been
jointly
reviewed
by
Health
Canada's
Pest
Management
Regulatory
Agency
(PMRA)
and
the
U.
S.
Environmental
Protection
Agency
(EPA).
Plant
Products
Co.
was
granted
a
temporary
registration
(Section
17)
in
Canada
on
June
3,
2002
for
use
of
P.
flocculosa
strain
PF­
A22
UL,
and
its
end­
use
product,
SPORODEX
L.

SPORODEX
L
is
a
biological
fungicide,
containing
1.
3%
(w/
w)
P.
flocculosa
strain
PF­
A22
UL,
intended
for
the
control
of
powdery
mildew
on
greenhouse
roses
and
cucumbers.
The
active
microorganism,
Pseudozyma
flocculosa,
is
a
naturally
occurring
fungus
and
is
not
currently
registered
in
the
U.
S.
or
Canada.

Microbial
pest
control
agents
are
increasingly
being
investigated
for
use
as
alternatives
to
conventional
pesticides
because
they
are
thought
to
pose
a
lower
potential
risk
to
human
health
and
the
environment,
compared
with
conventional
pesticides.
SPORODEX
L
represents
a
potential
biological
replacement
for
chemical
fungicides.

The
active
ingredient,
Pseudozyma
flocculosa
strain
PF­
A22
UL,
and
the
formulated
product
SPORODEX
L,
for
control
of
powdery
mildew
on
greenhouse­
grown
roses
and
cucumbers
have
been
granted
temporary
registration
pursuant
to
Section
17
of
the
Pest
Control
Products
Regulations
(Canada)
and
a
conditional
registration
pursuant
to
Section
3(
c)(
7)(
C)
of
the
Federal
Insecticide
Fungicide
and
Rodenticide
Act
(FIFRA)
on
the
condition
that
confirmatory
data
are
submitted.

A
summary
of
PMRA's
and
EPA's
findings
in
support
of
this
decision
is
found
in
this
Monograph.
A
copy
of
PMRA's
Sporodex
Regulatory
Note
can
be
found
on
the
PMRA
internet
site
at
the
following
address:
http://
www.
hc­
sc.
gc.
ca/
pmra­
arla/
english/
pdf/
reg/
reg2002­
02­
e.
pdf.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
3
Table
of
Contents
Chapter
1
Theactivemicro­
organism,
itspropertiesanduses................
7
1.
1
Identity
of
the
active
micro­
organism
and
preparation
containing
it
.
.
.
7
Table1.
1­
1
TGAIIdentification
............................
7
1.2
Physical
and
chemical
properties
of
technical
and
end­
use
product(
s)
.
.
8
Table
1.
2­
1
Technical
Product:
Pseudozyma
flocculosa
strain
PF­
A22UL.............................................
8
Table1.
2­
2
End­
UseProduct:
SPORODEXL..................
8
1.
3
Detailsofusesandfurtherinformation.........................
8
Chapter
2
Methodsofanalysis.......................................
9
2.
1
Methodsfor
analysisofthemicro­
organismasmanufactured........
9
2.
1.
1
Methodsfor
identificationofthemicro­
organism.................
9
2.
1.
2
Methodsfor
establishmentofpurityofseedstock
...............
10
2.
1.
3
Methods
to
define
the
content
of
the
micro­
organism
in
the
manufactured
material
used
for
the
production
of
formulated
products
.....................................................
11
2.1.4
Methods
for
the
determination
of
relevant
impurities
in
the
manufactured
material...............................................
11
2.
1.
5
Methods
to
show
absence
of
any
human
and
mammalian
pathogens
.
.
12
2.
1.
6
Methods
to
determine
storage
stability,
shelf­
life
of
the
micro­
organism
.....................................................
12
2.
2
Methods
to
determine
and
quantify
residues
(viable
or
non­
viable)
of
the
activemicro­
organismandrelevantmetabolites
.................
12
Chapter
3
Impactonhumanhealthandsafety
..........................
12
3.
1
Toxicityandinfectivitysummaries(
Tier
I
acutestudies)
..........
13
3.
1.
1
Acuteoraltoxicity/
pathogenicitystudy
......................
13
3.
1.
2
Acutepulmonarytoxicity/
pathogenicitystudy
.................
13
3.
1.
3
Acutepulmonaryrange­
findingstudy.........................
14
3.
1.
4
Intraperitonealtoxicity/
infectivitystudy......................
15
3.
1.
5
Acutedermaltoxicity/
irritationstudy........................
16
3.
1.
6
Primaryeyeirritationstudy
................................
16
3.
1.
7
Subchronic,
chronictoxicityandoncogenicity
..................
16
3.
1.
8
Effects
on
the
immune
and
endocrine
systems
..................
17
Table
3.
1
Summary
of
toxicity
and
pathogenicity
studies
with
Pseudozyma
flocculosa
.............................................
17
3.
1.
9
Integratedtoxicityandinfectivitysummary
....................
19
3.
2
Hypersensitivityincidents
.................................
21
3.3
Impact
on
human
and
animal
health
arising
from
exposure
to
the
active
substanceor
toimpuritiescontainedinit
......................
21
3.
3.
1
Occupationalandbystanderexposureassessment
...............
21
Chapter
4
Residues
..............................................
22
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
4
4.
1
Residuesrelevanttoconsumer
safety.........................
22
4.
1.
1
Dietaryexposureandriskassessment
........................
22
4.
1.
2
Drinkingwaterexposureandriskassessment...................
23
4.
1.
3
Maximum
residue
limits
...................................
23
4.2
Aggregate
exposure
from
multiple
routes
including
oral,
dermal,
and
inhalation..............................................
23
4.
2.
1
Oral..................................................
23
4.
2.
2
Dermal
...............................................
24
4.
2.
3
Inhalation
.............................................
24
4.
3
Cumulativeeffects.......................................
24
4.
4
DeterminationofsafetyforU.
S.
population,
infantsandchildren....
24
Chapter
5
Fateandbehaviourintheenvironment........................
25
Chapter
6
Effectsonnon­
targetspecies...............................
25
6.
1
Birds
.................................................
25
6.
1.
1
Avianoral
.............................................
25
6.
1.
2
Avianpulmonary/
inhalation/
injection.........................
25
6.
2
Wildmammals..........................................
26
6.
3
Fish..................................................
26
6.
3.
1
Freshwater
fishandestuarine/
marineanimals...................
26
6.
4
Arthropods
............................................
27
6.
4.
1
Terrestrialarthropods
....................................
27
6.
4.
2
Aquaticarthropods
......................................
27
6.
5
Non­
arthropodinvertebrates
...............................
27
6.
6
Microorganisms.........................................
28
6.
7
Plants
................................................
28
6.
7.
1
Aquaticplants
..........................................
28
6.
7.
2
Terrestrialplants
........................................
28
Table
6.
1
Risks
of
Pseudozyma
flocculosa
strain
PF­
A22
UL
to
non­
target
organisms
.............................................
29
6.
8
Integratedenvironmentaltoxicologysummary..................
30
Chapter
7
Efficacy
data
and
information
..............................
30
7.
1
Effectiveness
...........................................
30
7.
1.
1
Intendeduse
...........................................
30
7.
1.
2
Modeofaction
.........................................
31
7.
1.
3
Crops
................................................
31
7.
1.
4
Effectivenessagainstpest
.................................
31
7.
1.
5
Totalsprayvolume
......................................
32
7.
2
Phytotoxicity
to
target
plants
(including
different
cultivars),
or
to
target
plant
products
(OECD
7.
4)
....................................................................
33
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
5
7.3
Observations
on
undesirable
or
unintended
side
effects
e.
g.
on
beneficial
and
other
non­
target
organisms,
on
succeeding
crops,
other
plants
or
parts
of
treated
plants
used
for
propagating
purposes
(e.
g.
seed,
cutting,
runners)...............................................
33
7.3.1
Impact
on
succeeding
crops
................................
33
7.3.2
Impact
on
adjacent
crops
..................................
33
7.
3.
3
Impact
on
seed
viability
...................................
33
7.
4
Economics
.............................................
33
7.
5
Sustainability
...........................................
34
7.
5.
1
Surveyofalternatives
....................................
34
7.
5.
1.
1
Non­
chemicalcontrolpractices
.............................
34
7.
5.
1.
2
ChemicalControlPractices
................................
34
Table7.
5­
1
Alternativediseasecontrolproducts
...............
35
7.
5.
2
Compatibility
with
current
management
practices
including
IPM
....
35
7.
5.
3
Contributiontoriskreduction
..............................
35
7.
5.
4
Information
on
the
occurrence
or
possible
occurrence
of
the
development
ofresistance
...........................................
36
7.
6
Conclusions............................................
36
7.
6.
1
Summary..............................................
36
Table7.
6­
1
Summaryoflabelproposalsandrecommendations...............
37
Chapter
8
Overallconclusions
......................................
37
8.
1
Productcharacterizationandanalysis.........................
37
8.
2
Toxicityandinfectivity
...................................
38
8.
3
Exposure..............................................
38
8.
4
Foodandfeedresidues
...................................
38
8.
5
Environmentalassessment
.................................
39
8.6
Efficacy
assessment
......................................
39
Chapter
9
Riskmanagementconsiderations
............................
40
9.
1
Publicinterestfinding
....................................
40
9.
2
Determination
of
3(
c)(
7)(
C)
eligibility
........................
40
9.
3
Termsandconditionsofregistration
.........................
42
9.
4
Tolerance
.............................................
43
9.
5
Codexharmonization.....................................
43
9.
6
Riskmigitation
.........................................
43
9.
7
Endangeredspecies
......................................
44
9.
8
Labelsandlabeling
......................................
44
9.
8.
1
End­
useproduct
........................................
44
9.
8.
2
Manufacturing­
useproduct................................
50
Chapter
10
Listofabbreviations
.....................................
53
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
6
Product
Monograph
Team
U.
S.
EPA/
Office
of
Pesticide
Programs/
Biopesticides
and
Pollution
Prevention
Division:

Sharlene
Matten,
Ph.
D.
Biologist,
Regulatory
Action
Leader
Ibrahim
Barsoum,
Ph.
D.
Microbiologist,
Product
Characterization
and
Human
Health
Analysis
John
Kough,
Ph.
D.
Biologist,
Senior
Scientist
Zigfridas
Vaituzis,
Ph.
D.
Microbiologist,
Environmental
Fate
and
Effects
Analysis
Barbara
Mandula,
Ph.
D.
Biologist
Suzanne
Krolikowski,
J.
D.
Office
of
General
Counsel
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
7
Chapter
1
The
active
micro­
organism,
its
properties
and
uses
1.1
Identity
of
the
active
micro­
organism
and
preparation
containing
it
Table
1.
1­
1
TGAI
Identification
Active
Micro­
organism
Pseudozyma
flocculosa
strain
PF­
A22
UL
Function
Biological
fungicide
Binomial
name:
Pseudozyma
flocculosa
(Traquair,
J.
A.,
Shaw,
L.
A.,
and
Jarvis,
W.
R.)
Boekhout,
T.
andTraquair,
J.
A.
strainPFA22
UL
Taxonomic
designation:

Kingdom:
Phylum:

Genus:
Species:
Strain:
Fungi
Deuteromycotina
Dematiaceous
Asexual
Fungi
Pseudozyma
flocculosa
PF­
A22
UL
Nominal
purity
of
active
Pseudozyma
flocculosa
strain
PF­
A22
UL
(TGAI)
consists
of
100%
active
ingredient
in
spent
fermentation
medium
corresponding
to
a
minimum
of
3
×
10
8
colony
forming
units
(CFU)/
mL
of
Pseudozyma
flocculosa
strain
PF­
A22
UL.

1.3%
w/
w
(equivalent
to
a
min.
3
×
10
8
CFU/
mL)
in
SPORODEX
L
(EP).

Identity
of
relevant
impurities
of
toxicological,
environmental
and/
or
other
significance
A
stock
culture
is
rejected
if
biological
activity
is
altered
or
if
mutations
are
detected.
If
any
contamination
is
found
in
the
media
prior
to
inoculation,
the
media
is
discarded.
If
contamination
exceeds
the
product
release
standards
for
total
aerobic
flora
(<
1000
CFU/
mL),
enterobacteria
(<
10
CFU/
mL),
fecal
streptococci
(absence
in
1
gram),
Staphylococcus
aureus
(absence
in
1
gram),
coliforms
(<
10
CFU/
mL),
Escherichia
coli
(absence
in
1
gram)
and
Salmonella
(absence
in
1
gram),
the
product
is
discarded.
No
mammalian
toxins
are
known
to
be
produced
by
strain
PF­
A22
UL.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
8
1.2
Physical
and
chemical
properties
of
technical
and
end­
use
product(
s)

Table
1.
2­
1
Technical
Product:
Pseudozyma
flocculosa
strain
PF­
A22
UL
Not
applicable.
SPORODEX
L
is
manufactured
following
a
continuous
manufacturing
process
that
does
not
involve
an
intermediate
stand­
alone
technical
product.

Table
1.
2­
2
End­
Use
Product:
SPORODEX
L
Property
SPORODEX
L
Physical
state
at
25

C
Liquid
Colour
Beige
Odour
Faint
mushroom
smell
pH
in
distilled
water
6.
4­
6.
8
Density
1.
05
g/
mL
Viscosity
51
centipoise
Corrosion
character
None
All
formulants
in
Sporodex
L
are
either
of
food
grade
quality
or
are
considered
relatively
nontoxic
(i.
e.,
EPA
list
3,
4A
or
4B).

1.3
Details
of
uses
and
further
information
SPORODEX
L
is
an
end­
use
product
containing
the
active
ingredient
Pseudozyma
flocculosa
strain
PF­
A22
UL.
SPORODEX
L
is
a
liquid
proposed
for
use
as
a
biological
fungicide
to
control
powdery
mildew
fungi
(Sphaerotheca
pannosa
var.
rosae
and
Sphaerotheca
fuliginea)
on
greenhouse
food
and
non­
food
crops,
namely
cucumber
and
roses.
SPORODEX
L
is
to
be
applied
in
an
aqueous
solution
prepared
by
diluting
500
mL
of
product
per
100
L
of
water
(or
64
U.
S.
fl
oz
per
100
U.
S.
gallons
of
water)
(equivalent
to
approximately
10
5
to
10
6
CFU/
mL).
A
wetting
agent
is
added
to
a
final
concentration
of
0.
02%
to
improve
its
efficacy.
Plants
are
to
be
treated
beginning
when
environmental
conditions
favour
development
of
powdery
mildew
or
at
the
first
sign
of
the
disease.
Plants
are
to
be
sprayed
to
the
point
of
run­
off
at
weekly
intervals.
Up
to
1500
L
of
spray
solution
is
to
be
applied
per
hectare
(or
150
U.
S.
gallons
of
spray
mixture
per
acre)
for
cut
roses
or
cucumbers
or
about
1000
L/
ha
(or
100
U.
S.
gallons
per
acre)
for
potted
roses.
After
application,
the
relative
humidity
is
to
be
maintained
above
70%
for
12
hours.

Pseudozyma
flocculosa
was
isolated
in
1986
from
the
leaves
of
red
clover,
Trifolium
pratense,
infected
with
powdery
mildew,
Erysiphe
polygoni,
by
researchers
at
Agriculture
and
Agri­
Food
Canada,
Harrow,
Ontario.
Initially,
this
organism
was
erroneously
identified
as
a
new
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
9
ascomycetous
yeast
with
an
anamorphic
state
in
the
broad
genus
Sporothrix
and
a
teleomorphic
state
in
the
genus
Stephanoascus.
In
1995,
its
taxon
was
changed
to
Pseudozyma
flocculosa
following
ribosomal
DNA
analysis.
The
genus
Pseudozyma
contains
other
smut­
like
anamorphs,
including
P.
rugulosa
(formerly
Sporothrix
rugulosa).
Pseudozyma
flocculosa
is
a
phyllosphere
epiphyte
and
hyperparasite
of
primarily
powdery
mildew
but
has
been
isolated
in
association
with
other
leaf­
surface
moulds.
It
is
widely
distributed
in
North
America
(Canada
and
USA)
and
in
Europe
on
aerial
plant
surfaces
in
field
or
greenhouse
agricultural
ecosystems.

Pseudozyma
flocculosa
antagonizes
a
number
of
different
powdery
mildew
fungi
(Sphaerotheca
pannosa
var.
rosae,
Sphaerotheca
fulginea,
Erysiphe
graminis
var.
tritici
and
Erysiphe
polygoni)
on
many
different
plants
in
greenhouse
and
field
environments
when
the
relative
humidity
is
greater
or
equal
to
70%.
This
fungus
is
a
necrotroph
mycoparasite
that
kills
susceptible
target
host
cells
upon
contact
or
in
close
proximity.
Rapid
death
and
collapse
of
host
cells
without
penetration
is
brought
about
by
the
secretion
of
three
fungitoxic
unsaturated
C­
17
fatty
acids
(9­
heptadecenoic
acid,
6­
methyl­
9­
heptadecenoic
acid
and
4­
methyl­
7,
11­
heptadecadienoic
acid)
and
an
acyclic
norterpene
(2,
6,
10,
14,
18­
pentamethyl­
2,
6,
8,
10,
12,
14,
17­
nonadecaheptene­
1,
19­
diol).
The
fungitoxins
disrupt
susceptible
plasma
membranes
and
cytoplasmic
organelles
within
30
minutes
of
exposure.
The
inhibitory
response
includes
a
loss
of
proteins
and
electrolytes.
After
24
hours,
the
host
cells
rapidly
collapse
and
die
as
a
result
of
the
activity
of
the
fungitoxins
on
the
host
cell's
membranes
and
lipids.
Sensitivity
to
the
unsaturated
C­
17
free
fatty
acids
is
related
to
a
high
degree
of
unsaturation
of
phospholipid
fatty
acids
and
a
low
proportion
of
sterols.

Chapter
2
Methods
of
analysis
2.1
Methods
for
analysis
of
the
micro­
organism
as
manufactured
2.1.1
Methods
for
identification
of
the
micro­
organism
Appropriate
methodologies
for
detection,
isolation
and
enumeration
of
P.
flocculosa
strain
PFA22
UL
were
detailed
by
the
applicant.
The
microbial
pest
control
agent
(MPCA)
is
identified
using
a
combination
of
morphological
traits,
molecular
techniques
and
biological
activity.

The
identification
of
Pseudozyma
to
the
species
level
is
done
using
a
standard
mycological
approach.
Pseudozyma
species
can
be
differentiated
from
morphologically
similar
species
such
as
Hyalodendron,
Tilletiopsis,
Sporobolomyces
and
Sporothrix.
The
branching
conidiophores
of
Pseudozyma
can
be
confused
with
those
produced
by
Hyalodendron;
however,
the
whole
cell
hydrolysates
of
this
filamentous
basidiomycete
contain
xylose
which
is
not
found
in
Pseudozyma.
Tilletiopsis
and
Sporobolomyces,
other
saprophytic
wild
yeasts
on
aerial
plant
surfaces,
are
different
from
Pseudozyma
in
that
they
produce
spores
that
are
forcibly
discharged
upon
sporulation
(ballistospores).
Furthermore,
Tilletiopsis
species
produce
a
fungus­
degrading
 
­1,
3
glucanase
that
is
not
produced
by
Pseudozyma
species.
The
genus
Sporothrix
represents
a
group
of
anamorphic
ascomycetous
yeasts
such
as
Sporothrix
schenckii
(type),
an
animal
pathogen.
Physiologically,
Pseudozyma
species
differ
greatly
from
Sporothrix
species.
Unlike
the
ascomycetous
Sporothrix
anamorphs,
P.
flocculosa
shows
positive
reactions
in
Diazonium
Blue
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
10
B
and
urease
tests
typical
of
all
basidiomycetous
yeasts.
Also,
the
major
ubiquinone
is
Q­
10
rather
than
Q­
8
or
Q­
9
typical
of
the
ascomycetes,
Saccharomycopsis
and
Stephanoascus.

Strain
PF­
A22
UL
can
be
differentiated
from
other
strains
of
P.
flocculosa
using
a
DNA­
based
technique
called
multiplex
polymerase
chain
reaction
(multiplex
PCR).
The
multiplex
PCR
system
is
essentially
a
cocktail
of
different
primers
which
allows
the
rapid
assessment
of
numerous
DNA
fragments
in
a
single
PCR
amplification.
The
protocol
is
based
on
the
amplification
of
two
nuclear
regions,
(ITS
and
NS),
and
one
mitochondrial
region
(ML).
Those
regions
were
found
to
be
discriminant
in
the
identification
of
P.
flocculosa
PF­
A22
UL.

The
integrity
and
consistency
of
the
MPCA
is
ensured
by
two
methods.
The
first
method
is
a
DNA­
based
PCR
technique
called
random
amplified
microsatellites
PCR
(RAMS).
Microsatellites
are
hypervariable
non­
coding
regions
of
DNA
within
the
genome
that
evolve
more
rapidly
than
coding
DNA.
The
other
method
is
a
bioassay
that
measures
biological
activity.
The
biological
activity
of
the
MPCA
is
measured
by
the
inhibition
zone
created
when
a
susceptible
organism
is
grown
next
to
it.
Given
that
the
pest
controlled,
Sphaerotheca
species,
is
an
obligate
biotroph,
it
cannot
be
used
directly
in
this
bioassay.
Instead,
a
Phomopsis
species
is
used
because
its
sensitivity
to
P.
flocculosa's
fungitoxic
secretions
is
similar.

2.1.2
Methods
for
establishment
of
purity
of
seed
stock
The
mother
colony
is
maintained
as
slant
cultures
at
4

C,
and
as
freeze­
dried
cultures
stored
at
­20

C.
The
genetic
stability
of
those
cultures
is
verified
at
least
once
every
six
months
using
RAMS
PCR
(see
Section
2.
1.
1
for
details).
The
frequency
of
this
analysis
is
to
be
increased
accordingly
if
the
mother
colony
begins
to
show
signs
of
reduced
yield.

No
methods
for
establishing
the
purity
of
the
mother
colonies
were
submitted;
however,
sufficient
microbial
contaminant
screening
methods
were
proposed
for
the
production
of
Pseudozyma
flocculosa
strain
PF­
A22
UL
and
SPORODEX
L.
There
are
essentially
three
types
of
screening
methods
involved
in
the
production
of
Pseudozyma
flocculosa
strain
PF­
A22
UL
and
SPORODEX
L,
namely
pre­
fermentation
sterility
tests,
MPCA
integrity
tests,
and
microbial
contaminant
screening
tests.

Prior
to
inoculation,
all
media
are
screened
for
the
presence
of
microbial
contaminants
by
plating
aliquots
of
the
medium
onto
plate
count
agar
(PCA)
plates.
If
any
microbial
contamination
is
found,
the
medium
is
discarded.
Similarly,
all
cultures
are
monitored
for
MPCA
integrity
and
microbial
contamination
by
plating
various
dilutions
onto
potato
dextrose
agar
(PDA)
plates.
If
significant
microbial
contamination
is
detected,
the
culture
is
rejected.
In
case
of
abnormal
colony
morphology
on
PDA,
a
multiplex
PCR
analysis
(see
Section
2.
1.
1
for
details)
is
performed
to
properly
identify
the
afflicted
colonies.
Furthermore,
the
bioassay
method
described
in
Section
2.
1.
1
is
done
prior
to
product
formulation
to
verify
its
biological
control
potential.
Microbial
contaminant
screening
tests
are
performed
on
the
formulated
end­
use
product
prior
to
packaging.
They
are
monitored
by
culturing
dilutions
of
formulated
end­
use
products
onto
or
into
various
media.
The
groups
of
microbial
contaminants
tested
and
their
proposed
product
release
standards
include
total
aerobic
flora
(<
1000
CFU/
mL),
enterobacteria
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
11
(<
10
CFU/
mL),
fecal
streptococci
(absence
in
1
gram),
Staphylococcus
aureus
(absence
in
1
gram),
coliforms
(<
10
CFU/
mL),
Escherichia
coli
(absence
in
1
gram)
and
Salmonella
(absence
in
1
gram).
If
any
of
the
proposed
bioburden
limits
are
exceeded,
the
entire
batch
is
rejected.

2.1.3
Methods
to
define
the
content
of
the
micro­
organism
in
the
manufactured
material
used
for
the
production
of
formulated
products
The
concentration
of
Pseudozyma
flocculosa
strain
PF­
A22
UL
is
determined
by
measuring
the
number
of
viable
colony
forming
units
(CFU)
per
millilitre
of
formulated
product.
For
this
assay,
a
25­
mL
sample
is
diluted
in
peptone,
then
plated
onto
PDA.
Microscopic
observations
made
on
the
formulated
product
ensure
that
the
MPCA
is
under
the
proper
conidial
form.
According
to
product
specifications,
the
guarantee
is
expressed
as
greater
than
3
×
10
8
CFU/
mL.
The
biological
control
potential
of
the
MPCA
is
measured
prior
to
product
formulation
using
the
bioassay
method
described
in
Section
2.
1.
1.

2.1.4
Methods
for
the
determination
of
relevant
impurities
in
the
manufactured
material
No
known
or
suspected
toxic
material
is
produced
by
Pseudozyma
flocculosa
strain
PF­
A22
UL
during
the
fermentation
process.
Although
the
majority
of
the
manufacturing
process
is
designed
to
avoid
microbial
contamination,
some
contamination
can
occur
as
the
end­
use
product
is
centrifuged
and
formulated
under
non­
sterile
conditions.
As
mentioned
in
Section
2.
1.
2,
there
are
various
methods
to
monitor
the
levels
of
various
groups
of
contaminating
microorganisms
in
the
formulated
product.

Quality
control
data
from
five
batches
(1
commercial­
scale
and
4
pilot­
scale
batches)
of
SPORODEX
L
were
assessed
using
the
microbial
contaminant
screening
methods
described
in
Section
2.
1.
1.
The
total
aerobic
flora
in
SPORODEX
L
ranged
from
150
to
2
×
10
4
CFU/
mL.
Both
the
enterobacteria
and
fecal
coliform
counts
were
0
CFU/
mL
and
no
enterococci,
E.
coli,
Staphylococcus
aureus,
orSalmonella
were
detected
in
SPORODEX
L.
It
must
be
noted
that
two
of
the
five
batches,
including
the
only
commercial
batch,
were
destroyed
due
to
microbial
contamination.
In
one
of
those
batches,
the
total
aerobic
flora
exceeded
the
product
release
standard
for
this
group
of
contaminants,
i.
e.,
<
10
3
CFU/
mL.
In
the
other,
significant
microbial
contamination
was
detected
during
a
MPCA
integrity
test
on
PDA.
Both
of
those
batches
were
rejected.

Given
that
two
batches
were
destroyed
as
a
result
of
microbial
contamination,
the
submission
of
certificates
of
analysis
for
all
production
batches
of
SPORODEX
L
will
be
required
as
a
condition
of
registration
by
the
Canadian
Pest
Management
Regulatory
Agency
(PMRA)
and
the
U.
S.
Environmental
Protection
Agency
(EPA).
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
12
2.1.5
Methods
to
show
absence
of
any
human
and
mammalian
pathogens
As
discussed
in
Section
2.
1.
2,
the
quality
assurance
program
implemented
by
the
applicant
for
the
production
of
SPORODEX
L
requires
the
destruction
of
the
batch
if
any
of
those
product
release
standards
(including
animal
and
human
pathogens)
are
exceeded
in
the
formulated
product.

2.1.6
Methods
to
determine
storage
stability,
shelf­
life
of
the
micro­
organism
Storage
stability
data
are
required
to
ensure
product
performance
and
safety.
The
data
included
in
the
submission
package
were
derived
from
a
single
batch
of
SPORODEX
L
over
a
period
of
11
months
at
­20

C.
Additional
storage
stability
data
derived
from
at
least
five
production­
scale
or
pilot­
scale
batches
are
required
to
support
label
claims
and
ensure
product
performance
and
safety.
An
expiration
date
of
three
months
from
the
date
of
manufacture
is
required
until
additional
data
are
generated.

2.2
Methods
to
determine
and
quantify
residues
(viable
or
non­
viable)
of
the
active
micro­
organism
and
relevant
metabolites
Although
Pseudozyma
species
are
ubiquitous
in
nature
and
have
been
isolated
from
a
wide
variety
of
plant
surfaces
including,
leaf
litter,
clover,
maize
and
cucumbers,
no
adverse
effects
from
dietary
exposure
have
been
attributed
to
natural
populations
of
P.
flocculosa.
Given
that
there
are
no
significant
adverse
effects
reported
in
acute
oral
toxicity/
pathogenicity
study
and
that
there
are
no
reports
in
literature
suggesting
Pseudozyma
(Sporothrix)
flocculosa
produces
mammalian
toxins,
the
establishment
of
a
maximum
residue
limit
(MRL)
is
not
required
for
Pseudozyma
flocculosa
strain
PF­
A22
UL.
Consequently,
no
method(
s)
to
quantify
Pseudozyma
flocculosa
strain
PF­
A22
UL
residues
in
food
and
feed
are
required.

Analytical
methods
for
detecting
viable
Pseudozyma
flocculosa
residues
in
animal
and
human
body
tissues
involve
blending
of
tissues
and
recovery
on
yeast
malt
agar
(YM)
or
Martin's
agar
(MA).
If
needed,
a
multiplex
PCR
analysis
(see
Section
2.
1.
1
for
details)
can
be
performed
to
discriminate
strain
PF­
A22
UL
from
other
strains
of
P.
flocculosa.

Chapter
3
Impact
on
human
health
and
safety
P.
flocculosa
strain
PF­
A22
UL
was
considered
of
low
toxicity
and
no
pathogenicity
based
on
the
results
of
the
Tier
I
toxicology
studies.
Tier
II
and
Tier
III
studies
were
not
required
because
the
results
from
the
Tier
I
studies
were
sufficient
to
satisfy
guideline
requirements.
On
the
basis
of
the
studies
submitted,
it
was
considered
a
Toxicity
Category
III
pesticide
for
acute
oral
effects
due
to
the
amount
dosed
only,
and
Toxicity
Category
IV
for
dermal
and
primary
dermal
irritation
health
effects.
These
and
additional
toxicology
studies
are
summarized
below.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
13
3.1
Toxicity
and
infectivity
summaries
(Tier
I
acute
studies)

3.1.1
Acute
oral
toxicity
/
pathogenicity
study
(OPPTS
885.3050)
(MRID#
s
451152­
04
453634­
01)

No
signs
of
toxicity
or
pathogenicity
were
noted
when
SPORODEX
WP,
a
wettable
powder
formulation,
was
administered
to
rats
via
the
oral
route.

In
an
acute
oral
toxicity
study,
groups
of
fasted
6­
7
week
old
Fisher
344
rats
(12/
sex)
were
administered
a
single
oral
dose
of
SPORODEX
WP
in
USP
sterile
water
for
injection
at
doses
of
5.8
x
10
8
colony­
forming
units
(CFU)
per
animal
for
males
and
5.
6
x
10
8
CFU
per
animal
for
females.
An
equal
number
of
animals
were
dosed
with
heat­
killed
test
substance
and
four
animals/
sex
served
as
untreated
controls.
The
animals
were
then
observed
for
a
period
of
up
to
21
days
with
interim
scheduled
sacrifices.
No
effect
on
body
weight
gain
and
no
apparent
signs
of
treatment­
related
toxicity,
infectivity
or
pathogenicity
were
observed
in
any
of
the
treated
animals
during
the
study
period.
Clearance
of
the
test
organism
occurred
by,
or
prior
to,
posttreatment
day
7.
Based
on
the
results
of
this
study,
SPORODEX
L
and
its
active
ingredient,
P.
flocculosa,
is
not
considered
toxic
or
pathogenic
to
male
or
female
Fisher
344
rats.

The
test
substance
used
in
this
study
was
a
wettable
powder
formulation
of
SPORODEX.
A
change
in
the
intended
formulation
of
the
end­
use
products
from
a
wettable
powder
to
a
liquid
formulation
(SPORODEX
L),
however,
triggered
the
need
for
a
rationale
for
the
test
substance.
The
applicant
requested
a
waiver
from
submitting
a
replacement
acute
oral
study
using
the
TGAI
or
the
liquid
formulation
based
on
the
fact
that
the
new
formulants
found
in
SPORODEX
L
are
of
food
grade
quality
and
that
the
levels
of
other
formulants
have
been
significantly
reduced.
The
toxicity
of
the
liquid
formulation
is,
therefore,
expected
to
be
less
than
that
of
the
wettable
powder
formulation
that
was
tested.

3.1.2
Acute
pulmonary
toxicity
/
pathogenicity
study
(OPPTS
885.3150)
(MRID#
s
451152­
06,
453634­
01)

The
potential
toxicity
and
pathogenicity
of
P.
flocculosa
was
tested
by
observing
the
effects
following
a
single
intratracheal
instillation
of
3.
2
x
10
7
CFU
of
the
test
organism
(TS)
to
each
of
12
male
and
12
female
CD
rats.
An
equal
number
of
animals
were
treated
with
heat­
killed
test
substance
(KTS)
and
four
animals/
sex
served
as
untreated
controls.
Animals
were
observed
for
up
to
14
days
with
interim
scheduled
sacrifices.

A
total
of
15
rats
(3/
8
male
and
2/
8
female
TS­
dosed
rats
and
6/
8
male
and
4/
8
female
KTSdosed
rats)
died
on
days
2
and
3.
Laboured
respiration,
rough
hair
coat,
ocular
discharge
and
nasal
discharge
were
observed
in
both
TS­
and
KTS­
dosed
rats.
Hunched
posture
and
lethargy
were
also
observed
in
one
female
and
one
male
TS­
dosed
rat,
respectively.
The
presence
or
absence
of
clinical
symptoms
were
not
indicative
of
spontaneous
deaths.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
14
Due
to
the
large
number
of
spontaneous
deaths
and
a
number
of
missed
data
collections,
data
for
evaluating
effects
on
body
weights,
food
consumption
and
relative
organ
weight
were
limited.
At
the
end
of
the
14­
day
long
study,
administration
of
P.
flocculosa
did
not
have
a
statistically
significant
effect
on
body
weight.
Analyses
of
daily
food
consumption
and
relative
organ
weights
were
skewed
as
they
were
either
not
determined
or
did
not
include
animals
that
died
prior
to
their
scheduled
sacrifice
dates.

At
necropsy,
liver
lesions
and
lesions
and
enlargement
of
the
lung
and
spleen
were
observed
in
both
TS­
and
KTS­
dosed
rats.
Confluent
dark
areas
were
also
seen
in
the
kidneys
of
a
single
male
TS­
dosed
rat.
These
necropsy
findings
were
considered
consistent
with
the
method
of
dosing
and
the
body's
normal
immunological
response
to
a
foreign
substance.

Pseudozyma
flocculosa
was
detected
in
the
lungs
and
lymph
nodes
and
the
stomach
and
small
intestine
of
TS­
dosed
animals
only.
Counts
in
these
tissues
were
below
the
limit
of
detection
by
day
7.

Based
on
this
study,
P.
flocculosa
is
toxic,
but
not
infective
or
pathogenic,
at
the
dose
administered
when
introduced
by
the
intratracheal
route
to
male
and
female
CD
rats.
This
acute
pulmonary
study,
however,
was
originally
classified
as
unacceptable
due
to
major
deficiencies
in
the
collected
toxicity
data
and
a
possible
dosing
error,
as
indicated
by
the
presence
of
the
MPCA
in
the
stomach
and
small
intestines
on
the
day
of
dosing.
However,
there
was
relevant
pathogenicity
information
that
indicated
clearance
of
the
MCPA.
Thus,
this
study
is
considered
to
be
supplemental
because
it
provides
acceptable
information
regarding
infectivity/
pathogenicity;
however,
this
study
does
not
differentiate
the
cause
of
certain
mortalities
in
the
TS
and
KTS
treatments.
A
confirmatory
acute
pulmonary
toxicity
/
pathogenicity
study
using
the
TGAI
and
testing
of
the
sterile
filtrate
from
the
production
culture
will
therefore
be
required
to
provide
this
additional
information
as
a
condition
of
registration.

3.1.3
Acute
pulmonary
range­
finding
study
(OPPTS
885.3150)
(MRID#
s
451152­
07,
453634­
01)

In
order
to
determine
whether
the
test
substance
(in
both
its
viable
and
non­
viable
forms),
P.
flocculosa,
was
the
cause
of
the
deaths,
a
subsequent
acute
pulmonary
range­
finding
toxicity
study
was
conducted.
In
this
range­
finding
study,
groups
of
young
adult
CD
rats
(5/
sex/
dose
level)
were
exposed
by
the
intratracheal
route
to
P.
flocculosa
(4.
2
x
10
7
CFU/
mL)
in
ASTM
Type
1
water
at
doses
of
4.
2
x
10
7
,3.
4x10
7
,6.
8x10
6
and
3.
4
x
10
6
CFU/
animal.
Animals
were
then
observed
for
14
days.
There
were
no
mortalities
and
all
animals
gained
weight
during
the
study.
Rough
hair
coat
occurred
in
a
dose­
dependent
manner
with
all
5
animals/
sex
exhibiting
this
symptom
at
the
highest
dose
of
4.
2
x
10
7
CFU/
animal.
One
female
dosed
with
4.2
x
10
7
CFU
experienced
tremors,
closed
eyes
and
rough
hair
coat.
Pseudozyma
flocculosa
was
classified
as
being
of
slight
toxicity
(EPA
Toxicity
Category
IV)
based
on
adverse
effects
observed
in
some
test
animals.

This
acute
pulmonary
study
was
considered
supplemental.
According
to
USEPA
OPPTS
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
15
885.3150,
the
minimum
dose
is
10
8
units
of
the
MPCA
per
test
animal.
The
maximum
dose
level
used
in
this
study,
however,
was
only
4.2
x
10
7
CFU/
animal.
The
maximum
dose
level
used
in
this
study,
however,
was
only
4.2
x
10
7
CFU/
animal.
Furthermore,
infectivity
was
not
addressed;
however,
the
acute
pulmonary
toxicity/
pathogenicity
study
did
address
infectivity
sufficiently.
Consequently,
this
study
does
not
satisfy
the
guideline
requirement
for
an
acute
pulmonary
study
(OPPTS
885.3150)
in
the
rat.
EPA,
in
considering
the
two
studies
together,
believes
that
there
are
sufficient
data
with
which
to
determine
the
toxicity
and
pathogenicity
of
Pseudozyma
flocculosa.
As
any
potential
inhalation
risk
that
is
raised
by
these
studies
is
primarily
a
worker
risk,
EPA
is
requiring
that
a
respirator
be
worn
by
workers
to
limit
any
inhalation
exposures.
In
addition,
a
Restricted­
Entry
Interval
(REI)
of
4
hours
is
required
for
early
entry
postapplication
workers
or
other
persons
entering
treated
greenhouses.
Finally,
a
confirmatory
acute
pulmonary
toxicity
/
pathogenicity
study
using
the
TGAI
and
testing
of
the
sterile
filtrate
from
the
production
culture
will
be
required
as
a
condition
of
registration.

3.1.4
Intraperitoneal
toxicity
/
infectivity
study
(OPPTS
885.3200)
(MRID#
s
451152­
08,
453634­
01)

In
an
acute
intraperitoneal
toxicity/
infectivity
study,
groups
of
young
adult
CD
rats
(4/
sex/
scheduled
sacrifice
date)
were
exposed
by
the
intraperitoneal
route
to
an
undiluted
suspension
of
P.
flocculosa
(TS)
ata
dose
of3.
5x10
7
CFU/
animal
(in
1.
0
mL).
Animals
were
then
observed
for
up
to
14
days.
An
equal
number
of
young
adult
CD
rats
were
similarly
injected
with
heat­
killed
test
substance
(KTS).
An
undosed
naive
control
(NC)
group
consisting
of
4
rats/
sex
was
also
included
in
the
study.
Cage
side
observation
for
clinical
symptoms
was
performed
daily
and
animal
body
weights
and
food
consumption
were
monitored.

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

No
adverse
clinical
signs
were
observed
at
any
point
of
the
study
in
any
of
the
groups
of
rats.
Body
weight
gain
of
TS­
dosed
male
rats
was
significantly
decreased
while
this
group's
food
consumption
was
significantly
increased
compared
to
NC
animals.
There
was
no
significant
difference
between
KTS­
dosed
and
NC
animals
in
terms
of
body
weight,
body
weight
gain
or
food
consumption.
Upon
necropsy
of
TS­
and
KTS­
dosed
animals,
white
nodules
and
higher
relative
spleen
weights
were
observed
and
attributed
to
a
normal
immune
response
to
a
foreign
substance.
The
detection
of
P.
flocculosa
in
the
peritoneal
fluid
lavage
of
TS­
dosed
male
rats
was
consistent
with
the
method
of
administration.
Clearance
of
P.
flocculosa
from
all
other
tissues
and
fluids
occurred
by
day
7.
No
test
substance
was
detected
from
any
of
the
organs
of
the
KTS­
dosed
or
NC
animals.

At
the
dose
administered,
P.
flocculosa
was
slightly
toxic
but
not
pathogenic
to
male
and
female
CD
rats
when
introduced
by
the
intraperitoneal
route.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
16
3.1.5
Acute
dermal
toxicity
/
irritation
study
(
OPPTS
885.3100)
(MRID#
s
451152­
09,
453634­
01)

In
an
acute
dermal
toxicity
study,
a
single
group
of
New
Zealand
White
rabbits
(5/
sex)
was
dermally
exposed
to
1.
2
x
10
7
CFU
P.
flocculosa
(equivalent
to
approximately
0.82­
0.90
g/
kg
bw
for
males
and
0.
80­
0.91
g/
kg
bw
for
females),
for
24
hours
to
an
area
equivalent
to
approximately
10%
of
the
dorsal
skin
surface.
Following
exposure,
the
animals
were
observed
for
a
period
of
14
days.

No
treatment­
related
signs
of
toxicity
or
skin
irritation
were
observed
in
any
animal
during
the
14­
day
observation
period.
At
the
dose
administered,
P.
flocculosa
was
not
considered
toxic
or
irritating
to
the
skin.

The
recommended
test
substance
for
acute
dermal
toxicity
and
acute
dermal
irritation
studies
is
the
end­
use
product.
Instead,
the
test
substance
was
produced
by
the
test
facility
using
a
method
different
from
the
proposed
manufacturing
method.
An
acceptable
waiver
rationale
was
submitted
to
address
the
toxicity
and/
or
irritation
potential
of
the
formulation
ingredients.
The
waiver
rationale
was
based
on
the
formulation
ingredients
being
of
food­
grade
quality
or
considered
as
relatively
non­
toxic.

3.1.6
Primary
eye
irritation
study
(OPPTS
870.2400)
(MRID#
s
451152­
10,
453634­
01)

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

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

3.1.7
Subchronic,
chronic
toxicity
and
oncogenicity
Survival,
replication,
infectivity,
significant
toxicity
or
persistence
of
the
MPCA
was
not
observed
in
the
test
animals
treated
in
Tier
I
acute
oral,
pulmonary
and
intravenous
toxicity/
infectivity
tests.
Consequently,
higher
tier
tests
involving
subchronic
and
chronic
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
17
testing,
oncogenicity
testing,
mutagenicity
and
teratogenicity
were
not
required
based
on
the
lack
of
concerns
following
analysis
of
Tier
I
test
results.

3.1.8
Effects
on
the
immune
and
endocrine
systems
EPA
does
not
have
any
information
regarding
endocrine
effects
of
this
microbial
pesticide
at
this
time.
There
is
no
evidence
to
suggest
that
use
of
P.
flocculosa
strain
PF
A­
22
UL
at
the
proposed
concentrations
will
adversely
affect
the
endocrine
or
immune
systems.
The
active
ingredient,
P.
flocculosa
strain
PF­
A22
UL,
is
not
known
to
be
a
human
pathogen
nor
an
endocrine
disrupter.
The
submitted
toxicity/
pathogenicity
studies
in
the
rodent
indicate
that,
following
several
routes
of
exposure,
the
immune
system
is
still
intact
and
able
to
process
and
clear
the
active
ingredient.
Therefore,
no
adverse
effects
to
the
immune
and
endocrine
systems
are
known
or
expected.
Based
on
this
rationale,
the
registrant
waiver
request
for
OPPTS
880.3800
(Immune
Response)
was
found
to
be
acceptable.

Table
3.
1
Summary
of
toxicity
and
pathogenicity
studies
with
Pseudozyma
flocculosa
STUDY
SPECIES/
STRAIN
AND
DOSES
/
TEST
SUBSTANCE
LD50
,
MIS/
MAS
TARGET
ORGAN/
SIGNIFICANT
EFFECTS/
COMMENTS
ACUTE
STUDIES
Oral
(MRID#
s
451152­
04,
453634­
01)
Rat
­
Fisher
344,
12/
sex,


5.8
x
10
8
CFU
1
/animal

5.6
x
10
8
CFU/
animal
SPORODEX
WP

LD50
>5.
8×
10
8
CFU/
animal

LD50
>5.
6×
10
8
CFU/
animal
No
effect
on
body
weight
gain
or
feed
consumption
and
no
clinical
signs
of
treatment­
related
toxicity,
infectivity
or
pathogenicity.
No
mortalities.
Agent
cleared
from
the
gastrointestinal
tract
within
seven
days
of
dosing
and
was
not
detected
in
the
urine,
blood
or
other
organs
at
any
time.
No
significant
findings
observed
at
necropsy.
LOW
TOXICITY
AND
NO
PATHOGENICITY.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
18
Pulmonary
(MRID#
s
451152­
06
453634­
01)
Rat
­
CD,
12/
sex,
3.2
x
10
7
CFU/
animal
Pseudozyma
flocculosa
LD50
>3.
2×
10
7
CFU/
animal
Laboured
breathing,
rough
hair
coat,
ocular
discharge
and
nasal
discharge
observed
in
TS
2
­
and
KTS
3
­dosed
animals.
Hunched
posture
and
lethargy
observed
in
one
TS­
dosed
female
and
one
TS­
dosed
male,
respectively.
Mortalities
included
3

TS­
dosed,
6

KTS­
dosed,
2

TSdosed
and
4

KTS­
dosed
rats.
No
effect
on
body
weight
based
on
rats
sacrificed
on
day
14.
Daily
food
consumption
analysis
and
relative
organ
weights
either
not
determined
or
did
not
include
animals
that
died
prior
to
their
scheduled
sacrifice
dates.
Necropsy
findings
including
lesions
and
enlargement
of
the
lung,
confluent
dark
areas
in
the
kidneys,
lesions
and
enlargement
of
the
spleen
and
lung
lesions
in

and

rats
dosed
with
TS
and
KTS
were
attributed
to
the
method
of
dosing
and
the
body's
normal
immunological
response
to
a
foreign
substance.
Agent
was
detected
in
the
lungs
and
lymph
nodes,
stomach
and
small
intestines.
Clearance
from
these
organs
by
day
7.
Study
classified
as
SUPPLMENTAL.
This
study
is
considered
to
be
supplemental
because
it
provides
acceptable
information
regarding
infectivity/
pathogenicity;
however,
this
study
does
not
differentiate
the
cause
of
certain
mortalities
in
the
TS
and
KTS
treatments.
Confirmatory
acute
pulmonary
toxicity/
pathogenicity
study
is
required
using
the
TGAI
and
testing
of
the
sterile
filtrate
from
production
batches.

Pulmonary
Range
Finding
(MRID#
451152­
07
453634­
01)
Rat
­
CD,
5/
sex/
dose
level
4.2
x
10
7
CFU/
animal
3.4
x
10
7
CFU/
animal
6.8
x
10
6
CFU/
animal
3.4
x
10
6
CFU/
animal
Pseudozyma
flocculosa
LD50
>4.2
x
10
7
CFU/
animal
No
mortalities.
All
animals
gained
weight
over
the
course
of
the
14­
day
study.
Rough
haircoatoccurred
in
a
dosedependent
manner.
One
female
rat
dosed
at4.2
x
10
7
CFU
presented
with
tremors,
closed
eyes
and
rough
hair
coat.
SLIGHTLY
TOXIC;
PATHOGENICITY
NOT
DETERMINED.
Study
classified
as
SUPPLEMENTAL.
Upgraded
label
statements
required.
Confirmatory
acute
pulmonary
toxicity/
pathogenicity
study
is
required
using
the
TGAI
and
testing
of
the
sterile
filtrate
from
production
batches.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
19
Intraperitoneal
Injection
(MRID#
s
451152­
08
453634­
01)
Rat
­
Sprague
Dawley,
12/
sex,
3.5
x
10
7
CFU/
animal
Pseudozyma
flocculosa
LD50
>3.
5×
10
7
CFU/
animal
No
effect
on
body
weight
but
body
weight
gain
significantly
lower
in
TS­
dosed

rats
despite
increased
food
consumption
by
TS­
dosed

rats.
No
clinical
symptoms
or
mortalities.
White
nodules
noted
on
stomach,
caecum,
liver
or
small
intestine
of

and

rats
dosed
with
TS
and
KTS
attributed
to
normal
immunological
response
to
a
foreign
substance.
Increased
relative
spleen
weight
in

TS­
and
KTSdosed
rats
also
considered
to
be
a
normal
response.
Following
injection,
the
test
microbe
was
recovered
from
the
caecum,
kidneys,
liver,
lungs
and
associated
lymph
nodes,
spleen
and
stomach
and
small
intestines
of

and

TS­
dosed
rats.
Clearance
of
the
test
organism
occurred
within
7
days
of
administration.
SLIGHTLY
TOXIC
AND
NO
PATHOGENICITY
Dermal
Toxicity
and
Irritation
(MRID#
s
451152­
09,
453634­
01)
Rabbit
­
New
Zealand
White,
5/
sex
1.2
x
10
7
CFU/
animal
Pseudozyma
flocculosa
(equivalent
to
approximately
0.
82­
0.90
g/
kg
bw
for

and
0.
80­
0.91
g/
kg
bw
for

)
LD50
>1.
2×
10
7
CFU/
animal
(

LD50
>
0.
82­
0.90
g/
kg
bw

LD50
>
0.
80­
0.91
g/
kg
bw)
No
mortalities.
One

rabbit
lost
weight
within
the
first
week
but
experienced
a
slight
weight
gain
thereafter.
All
other
animals
gained
weight.
Slight
diarrhea
observed
in
one

7
days
after
administration.
No
other
adverse
clinical
symptoms.
No
signs
of
dermal
irritation.
LOW
TOXICITY
AND
NONIRRITATING

Eye
Irritation
(MRID#
s
451152­
10,
453634­
01)
Rabbit
­
New
Zealand
White,
6
females,
0.1
g
(equivalent
to
5.7
x
10
7
CFU/
animal)
SPORODEX
WP
MIS
4
=
1.
7
/
110
at
the
onehour
scoring
interval
MAS
5
=0.
22
Slight
conjunctival
redness
observed
in
5/
6
animals
at
the
one­
hour
scoring
interval.
By
the
24­
hour
scoring
interval,
only
2/
6
animals
continued
to
exhibit
slight
conjunctival
redness.
All
signs
of
ocular
irritation
were
absent
at
the
48­
hour
scoring
interval.
No
other
signs
of
ocular
irritation
or
adverse
clinical
symptoms.
No
mortalities.
SPORODEX
WP
formulation
expected
to
be
more
irritating
to
the
eye
than
SPORODEX
L.
MINIMALLY
IRRITATING.

1
CFU
=
Colony
Forming
Units
2
TS
=
Test
Substance
3
KTS
=
Killed
Test
Substance
4
MIS
=
Maximum
Irritation
Score
5
MAS
=
Maximum
Average
Score
(based
on
scores
from
24­,
48­
and
72­
hour
scoring
intervals)

3.1.9
Integrated
toxicity
and
infectivity
summary
The
registration
package
submitted
by
Plant
Products
Co.
in
support
of
registering
the
technical
grade
active
ingredient
(TGAI)
Pseudozyma
flocculosa
strain
PF­
A22
UL
and
the
end­
use
product
(EP)
SPORODEX
L,
was
reviewed
from
the
viewpoint
of
human
health
and
safety
and
was
determined
to
be
sufficiently
complete
to
permit
a
decision
on
registration.
The
information
provided
to
address
the
characterization
of
the
active
ingredient
as
well
as
the
manufacturing
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
20
process
and
quality
control
adequately
addressed
the
potential
human
health
and
safety
concerns
associated
with
P.
flocculosa
strain
PF­
A22
UL
and
bacterial/
fungal
contaminants
introduced
during
production.

No
signs
of
toxicity
or
pathogenicity
were
noted
when
SPORODEX
WP,
a
wettable
powder
formulation,
was
administered
to
rats
via
the
oral
route.

Intratracheal
administration
of
P.
flocculosa
resulted
in
a
significant
number
of
spontaneous
deaths
among
both
TS­
and
KTS­
dosed
animals.
Presence
of
the
test
organism
in
the
stomach
and
small
intestines
indicated
a
potential
dosing
error.
In
a
second
pulmonary
study,
there
were
no
mortalities
but
rough
hair
coat
occurred
in
a
dose­
dependent
manner.
Based
on
this
study,
P.
flocculosa
was
classified
as
slightly
toxic.
This
study
was
considered
supplemental
because
infectivity
was
not
assessed
and
because
the
test
substance
was
not
the
recommended
TGAI.
The
label
must
be
upgraded
with
a
statement
requiring
respirators
for
all
users
and
a
complete
acute
pulmonary
toxicity
/
infectivity
study,
using
the
TGAI,
will
be
required.

Pseudozyma
flocculosa
was
found
to
be
slightly
toxic
but
non­
pathogenic
when
administered
to
rats
via
intraperitoneal
injection.
There
were
no
mortalities
or
adverse
clinical
symptoms.
White
nodules
and
higher
relative
spleen
weights
were
noted
at
necropsy
and
attributed
to
a
normal
immune
response
to
a
foreign
substance.
Male
TS­
dosed
rats,
however,
exhibited
decreased
body
weight
gain
despite
increased
food
consumption
indicating
that
P.
flocculosa
was
slightly
toxic.
Clearance
of
the
test
organism
occurred
within
7
days
indicating
lack
of
pathogenicity.

P.
flocculosa
was
not
toxic
or
irritating
when
applied
dermally
to
rabbits.
A
waiver
rationale
was
submitted
to
address
the
toxicity
and/
or
irritation
potential
of
the
formulation
ingredients
in
SPORODEX
L.
All
formulation
ingredients
are
either
of
food­
grade
quality
or
classified
as
relatively
non­
toxic.
One
formulation
ingredient
may
cause
irritation
of
the
skin
with
prolonged
contact.
Standard
personal
protective
equipment
requirements
are
adequate.

Slight
conjunctival
redness
was
observed
after
administration
of
SPORODEX
WP
to
the
eyes
of
rabbits.
The
irritation
potential
of
SPORODEX
L
is
expected
to
be
less
than
that
of
SPORODEX
WP.
Standard
label
statements
instructing
users
to
avoid
contact
with
eyes
are
sufficient.

Pseudozyma
flocculosa
has
not
been
reported
to
produce
any
mammalian
toxins.
The
applicant
included
computer
literature
search
results
to
a
number
of
keywords
such
as
pseudozyma*,
tilletiopsis,
fate,
non
target,
carcin*,
mutagen*,
toxic*,
pathogen*,
antibiotic*,
polyen*,
sporothrix,
sporobolomyces,
rhodotorula,
phyllosphere
yeast*,
carcinog*
and
teratogen*.
The
literature
search
covered
AGRICOLA,
Biological
Abstracts,
CAB
Abstracts,
CHEMTOX,
RTEX
and
AGRIS
databases
from
1980
to
1999.
No
reports
of
mammalian
toxicity
were
found
in
standard
biological,
chemical
and
toxicological
abstracts.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
21
3.2
Hypersensitivity
(derminal
sensitization
study
OPPTS
870.2600
and
reports
of
incidents
OPPTS
885.3400)

The
applicant
has
also
submitted
an
acceptable
waiver
rationale
from
conducting
a
dermal
sensitization
study
based
on
the
assumption
that
most
microorganisms
contain
substances
that
could
elicit
a
hypersensitivity
response.
Pseudozyma
flocculosa
is
considered
a
potential
sensitizing
agent,
therefore,
the
statement,
"POTENTIAL
SENSITIZER"
is
required
on
the
principal
display
panels
of
the
technical
and
end­
use
formulation
labels.
The
use
of
personal
protective
equipment
will
also
be
required
to
mitigate
against
potential
dermal
sensitization
in
occupationally
exposed
workers/
handlers.

Skin
sensitizing
studies
are
not
considered
substitutes
for
timely
reports
of
hypersensitivity
incidents
subsequent
to
registration
approval.
No
adverse
effects
have
been
noted
among
researchers
who
have
worked
closely
with
P.
flocculosa
strain
PF­
A22
UL
for
up
to
10
years.
The
applicant
will
be
expected
to
report
any
subsequent
findings
of
hypersensitivity
or
other
health
incidents
to
workers,
applicators,
or
bystanders
exposed
to
the
MPCA
as
a
condition
of
registration.
Incident
reports
are
to
include
details
such
as
a
description
of
the
MPCA
and
formulation,
frequency,
duration
and
routes
of
exposure
to
the
material,
clinical
observations,
and
any
other
relevant
information.

3.3
Impact
on
human
and
animal
health
arising
from
exposure
to
the
active
substance
or
to
impurities
contained
in
it
3.3.1
Occupational
and
bystander
exposure
assessment
When
handled
according
to
the
label
instructions,
the
pulmonary,
dermal
and
ocular
routes
are
potential
routes
of
applicator
and
bystander
exposure.
Occupational
exposure
is
of
particular
concern
as
the
product
will
be
used
in
an
enclosed
environment.

U.
S.
EPA
and
Canada/
PMRA,
however,
do
not
expect
that
occupational
exposures
will
pose
an
undue
risk
on
the
basis
of
the
low
toxicity/
pathogenicity
profile.
While
submitted
acute
pulmonary
toxicity/
infectivity
studies
were
found
to
be
lacking,
inhalation
exposure
is
not
of
concern
if
the
required
respirator
is
worn
by
workers.
To
mitigate
dermal
and
inhalation
exposure
and
risk
to
workers,
use
of
appropriate
Personal
Protective
Equipment
(PPE)
will
be
required.
Furthermore
a
Restricted­
Entry
Interval
(REI)
of
4
hours
is
required
for
early
entry
(post­
application)
workers
or
other
persons
entering
treated
greenhouses.

Assuming
that
most
microorganisms
contain
substances
that
would
elicit
positive
hypersensitivity
reactions,
P.
flocculosa
strain
PF­
A22
UL
is
considered
a
potential
sensitizing
agent,
and
a
"POTENTIAL
SENSITIZER"
statement
will
be
required
on
the
principal
display
panel
of
the
TGAI
and
end­
use
formulation
labels.

The
label
does
not
allow
applications
to
turf,
residential
or
recreational
areas.
Because
the
use
sites
are
in
greenhouses,
exposure
to
infants
and
children
in
school,
residential
and
daycare
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
22
facilities
is
likely
to
be
minimal
to
non­
existent.
Consequently,
the
health
risk
to
infants
and
children
is
expected
to
be
negligible
to
non­
existent.

Chapter
4
Residues
In
examining
aggregate
exposure,
FFDCA
section
408
directs
U.
S.
EPA
to
consider
available
information
concerning
exposures
from
the
pesticide
residue
in
food
and
all
other
nonoccupational
exposures,
including
drinking
water
from
ground
water
or
surface
water
and
exposure
through
pesticide
use
in
gardens,
lawns,
or
buildings
(residential
and
other
indoor
uses).

Based
on
the
data
and
analyses
outlined
above,
U.
S.
EPA
has
concluded
that
there
is
a
reasonable
certainty
that
no
harm
will
result
from
aggregate
exposure
to
the
U.
S.
population,
including
infants
and
children,
to
residues
of
P.
flocculosa
strain
PF­
A22
UL
arising
from
use
on
greenhouse­
grown
cucumbers
and
roses.
This
includes
all
anticipated
dietary
exposures
and
all
other
exposures
for
which
there
is
reliable
information.

4.1
Residues
relevant
to
consumer
safety
4.
1.
1
Dietary
exposure
and
risk
assessment
The
proposed
food
use
pattern
is
likely
to
result
in
residues
in
or
on
food
and
feed.
Residues
of
the
microbial
pesticide
are
likely
to
be
removed
from
treated
food
by
washing,
peeling,
cooking
and
processing.
Even
if
residues
are
not
removed,
however,
EPA
believes
that
dietary
exposure
to
the
microbial
agent
will
result
in
negligible
to
no
risk
to
consumers.
Although
Pseudozyma
species
are
ubiquitous
in
nature
and
have
been
isolated
from
a
wide
variety
of
plant
surfaces
including
leaf
litter,
clover,
maize
and
cucumber,
no
adverse
effects
from
dietary
exposure
have
been
attributed
to
natural
populations
of
Pseudozyma
flocculosa.
Furthermore,
no
adverse
effects
were
observed
at
maximum
hazard
dose
levels
in
the
acute
oral
toxicity
/
pathogenicity
study
and
there
are
no
reports
of
known
mammalian
toxins
being
produced
by
the
MPCA.
Subchronic
and
chronic
dietary
exposure
studies
were
not
required
because
the
Tier
I
acute
oral
study
demonstrated
a
low
level
of
toxicity
and
no
pathogenicity
potential
for
the
active
microorganism.
Because
of
the
low
toxicity
profile
and
low
potential
exposure
of
the
MPCA
expected
for
the
proposed
uses,
there
is
no
concern
for
chronic
risks
posed
by
dietary
exposure
for
the
general
population
or
sensitive
subpopulations,
such
as
infants
and
children.
In
addition,
an
extensive
literature
search
yielded
no
reports
of
mammalian
toxins
being
produced
by
P.
flocculosa
(see
section
3.
1.
9).
The
fungitoxic
unsaturated
C­
17
fatty
acids
and
acyclic
norterpene
produced
by
the
MPCA
have
not
been
reported
to
be
toxic
to
mammals.
Neither
this
organism
nor
its
close
relatives
are
listed
among
microbial
contaminants
of
food.
Therefore,
EPA
expects
negligible
to
no
dietary
risk
from
exposure
to
naturally­
occurring
and
isolated
P.
flocculosa
strain
PF­
A22
UL
residues.

4.1.2
Drinking
water
exposure
and
risk
assessment
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
23
Although
heavy
rainfall
likely
carries
P.
flocculosa
into
neighboring
aquatic
environments,
growth
and
survival
of
terrestrial
fungi
such
as
P.
flocculosa
is
limited
in
such
environments.
Thus,
it
is
not
expected
to
proliferate
in
aquatic
habitats
following
incidents
of
direct
or
indirect
exposure
(e.
g.,
runoff
from
treated
greenhouses).
Moreover,
P.
flocculosa
is
not
considered
to
be
a
risk
to
drinking
water
because
of
minimal
to
non­
existent
toxicity.
Accordingly,
drinking
water
is
not
specifically
screened
for
P.
flocculosa
as
a
potential
indicator
of
microbial
contamination
or
as
a
direct
pathogenic
contaminant.
Both
percolation
through
soil
and
municipal
treatment
of
drinking
water
would
reduce
the
possibility
of
significant
transfer
of
residues
to
drinking
water.
Therefore,
the
potential
of
exposure
and
risk
to
humans
via
drinking
water
is
likely
to
be
minimal
to
non­
existent
for
this
MPCA.

4.1.3
Maximum
residue
limits
Although
Pseudozyma
species
are
ubiquitous
in
nature
and
have
been
isolated
from
a
wide
variety
of
plant
surfaces
including
leaf
litter,
clover,
maize
and
cucumber,
no
adverse
effects
from
dietary
exposure
have
been
attributed
to
natural
populations
of
Pseudozyma
flocculosa.
Furthermore,
no
adverse
effects
were
observed
in
the
acute
oral
toxicity
/
pathogenicity
study
and
there
are
no
reports
of
known
mammalian
toxins
being
produced
by
the
MPCA.
Therefore,
the
establishment
of
a
tolerance
or
maximum
allowable
residue
limit
is
not
required
for
P.
flocculosa
strain
PF­
A22
UL
under
Section
408
of
the
Federal
Food
Drug
and
Cosmetic
Act.

4.2
Aggregate
exposure
from
multiple
routes
including
oral,
dermal,
and
inhalation
The
current
label
does
not
allow
applications
to
turf,
residential
or
recreational
areas.
Because
the
use
sites
are
in
greenhouses,
exposure
to
the
U.
S.
population
including
infants
and
children
in
school,
residential
and
daycare
facilities
is
likely
to
be
minimal
to
non­
existent.
Consequently,
the
health
risk
posed
by
P.
flocculosa
strain
PF
A­
22
UL
from
non­
occupational
dermal
and
inhalation
exposures
to
the
general
public,
including
infants
and
children,
is
expected
to
be
negligible
to
non­
existent.
Any
concerns
for
potential
inhalation
risk
is
for
occupational
exposures,
and
as
mentioned
previously,
will
be
mitigated
by
the
requirement
of
a
respirator
and
restriction
of
the
reentry
interval.

4.2.1
Oral
Oral
exposure
would
occur
primarily
from
eating
treated
foods
and
from
drinking
water.
Residues
of
the
active
microorganism
can
be
easily
removed
from
treated
commodities
by
washing,
cooking,
peeling
and
processing.
Transfer
of
the
active
microogranism
to
drinking
water
is
not
likely
as
discussed
previously.
Thus
dietary
exposure
and
risk
are
likely
to
be
minimal
to
non­
existent.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
24
4.2.2
Dermal
Non­
occupational
dermal
exposure
and
risk
to
adults,
infants
and
children
are
not
likely
if
the
pesticide
is
used
as
labeled.
The
label
does
not
allow
applications
to
turf,
residential
or
recreational
areas.
The
only
use
sites
are
for
greenhouse­
grown
cucumbers
and
roses.

Dermal
exposure
via
the
skin
would
be
the
primary
route
of
exposure
for
applicators.
Since
unbroken
skin
is
a
natural
barrier
to
microbial
invasion
of
the
human
body,
dermal
absorption
could
occur
only
if
the
skin
were
cut,
if
the
microbe
were
a
pathogen
equipped
with
mechanisms
for
entry
through
or
infection
of
the
skin,
or
if
metabolites
were
produced
that
could
be
dermally
absorbed.
P.
flocculosa
is
not
known
to
be
a
human
pathogen
nor
is
it
known
to
produce
metabolites
that
are
dermally
absorbed.
Based
on
the
minimal
adverse
effects
in
the
intraperitoneal
study,
it
is
PMRA's
and
EPA's
opinion
that
even
cut
skin
should
not
pose
a
significant
risk
to
health
via
entry
of
absorbed
P.
flocculosa
into
the
body.

Although
the
MPCA
has
been
found
to
be
non­
toxic
and
non­
irritating
following
dermal
exposure,
it
is
a
potential
sensitizer.
Label
restrictions
and
risk
mitigation
measures
are
required
to
protect
populations
who
are
likely
to
be
primarily
exposed
to
the
pesticide.
Such
exposure
to
pesticide
handlers
can
be
ameliorated
if
they
wear
long­
sleeved
shirts,
long
pants,
shoes
and
socks.

4.2.3
Inhalation
Inhalation
would
be
another
route
of
exposure
for
mixer/
loader
applicators
and
possibly
earlyentry
workers.
Based
on
the
results
of
the
pulmonary
study
in
which
lesions
were
noted
on
the
lungs
of
some
treated
animals,
pesticide
handlers
must
wear
a
dust/
mist
filtering
respirator
with
the
NIOSH
prefix
N­
95,
R­
95,
P­
95
or
HE
filter
for
biological
products.

4.3
Cumulative
effects
The
Agency
has
considered
available
information
on
the
cumulative
effects
of
such
residues
and
other
substances
that
have
a
common
mechanism
of
toxicity.
These
considerations
included
the
cumulative
effects
on
infants
and
children
of
such
residues
and
other
substances
with
a
common
mechanism
of
toxicity.
EPA
is
not
aware
of
any
other
bacteria
or
other
substances,
besides
naturally­
occurring
strains
of
Pseudozyma,
that
share
a
common
mechanism
of
toxicity
with
this
active
ingredient.
Given
the
low
toxicity
and
pathogenicity
profile
of
P.
flocculosa,
even
if
there
were
any
other
substances
with
which
P.
flocculosa
shared
a
common
mechanism
of
toxicity,
no
adverse
cumulative
effects
are
expected.

4.4
Determination
of
safety
for
U.
S.
population,
infants
and
children
Based
on
the
toxicology
data
submitted
and
other
relevant
information
in
the
Agency's
files,
there
is
reasonable
certainty
no
harm
will
result
from
aggregate
exposure
of
residues
of
Pseudozyma
flocculosa
strain
PF­
A22
UL
to
the
U.
S.
population,
including
infants
and
children,
under
reasonably
foreseeable
circumstances
when
the
microbial
pesticide
product
is
used
as
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
25
labeled.
This
includes
all
anticipated
dietary
exposures
and
all
other
exposures
for
which
there
is
reliable
information.
The
Agency
has
arrived
at
this
conclusion
based
on
data
submitted
demonstrating
low
toxicity
at
the
maximum
doses
tested
and
a
lack
of
information
showing
adverse
effects
from
exposure
to
naturally
occurring
P.
flocculosa
as
well
as
a
consideration
of
the
product
as
currently
registered
and
labeled.
As
a
result,
EPA
establishes
an
exemption
from
tolerance
requirements
pursuant
to
FFDCA
408(
c)
and
(d)
for
residues
of
Pseudozyma
flocculosa
strain
PF­
A22
UL
in
or
on
all
food
commodities.

FFDCA
section
408
provides
that
EPA
shall
apply
an
additional
tenfold
margin
of
exposure
(safety)
for
infants
and
children
in
the
case
of
threshold
effects
to
account
for
prenatal
and
postnatal
toxicity
and
the
completeness
of
the
data
base
unless
EPA
determines
that
a
different
margin
of
exposure
(safety)
will
be
safe
for
infants
and
children.
Margins
of
exposure
(safety)
are
often
referred
to
as
uncertainty
(safety)
factors.
In
this
instance,
based
on
all
the
available
information,
the
Agency
concludes
that
P.
flocculosa
strain
PF­
A22
UL
is
practically
non­
toxic
to
mammals,
including
infants
and
children.
Thus,
there
are
no
threshold
effects
of
concern
and,
as
a
result
the
provision
requiring
an
additional
margin
of
safety
does
not
apply.
Further,
the
provisions
of
consumption
patterns,
special
susceptibility,
and
cumulative
effects
do
not
apply.
As
a
result,
EPA
has
not
used
a
margin
of
exposure
(safety)
approach
to
assess
the
safety
of
P.
flocculosa
strain
PF­
A22
UL.

Chapter
5
Fate
and
behavior
in
the
environment
Environmental
fate
data
(Tier
II)
were
not
triggered
as
adverse
effects
on
non­
target
organisms
are
not
expected
from
the
proposed
greenhouse
use
of
P.
flocculosa
strain
PF­
A22UL.
Environmental
fate
data
waiver
requestsby
Plant
Products
Co.
Ltd.
were
found
to
be
acceptable
(MRID#
451152­
11).

Chapter
6
Effects
on
non­
target
species
6.1
Birds
6.1.1
Avian
oral
6.1.2
Avian
pulmonary/
inhalation/
injection
(OPPTS
885.4050
and
OPPTS
885.4100,
MRID#
451152­
12)

Plant
Products
Co.
Ltd.
submitted
an
acceptable
justification
for
a
data
waiver
from
avian
oral
toxicity
studies
and
avian
pulmonary/
inhalation/
injection
studies.
The
waiver
request
was
based
on
the
rationale
that
P.
flocculosa
is
a
naturally­
occurring
soil
microorganism
whose
level
in
the
environment
will
not
significantly
increase
with
greenhouse
use
of
SPORODEX
L
and
that
an
extensive
literature
search
yielded
no
reports
of
adverse
effects
in
birds.

Use
of
SPORODEX
L
in
commercial
greenhouses
is
not
expected
to
result
in
increased
exposure
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
26
or
adverse
effects
in
birds.
Birds
will
not
be
directly
exposed
to
the
product
at
the
time
of
application.
Greenhouse
practices
designed
to
limit
exposure
to
the
outside
environment
will
limit
post­
application
exposure
to
birds.
Furthermore,
while
the
body
temperature
of
duck
and
quail
species
is
approximately
40

C,
Pseudozyma
species
do
not
grow
at
temperatures
beyond
37

C.
Consequently,
testing
is
considered
unnecessary
to
assess
the
risks
of
SPORODEX
L
to
avian
wildlife.

6.2
Wild
mammals
(OPPTS
885.4150,
MRID#
451152­
12)

Plant
Products
Co.
Ltd.
submitted
an
acceptable
justification
for
a
data
waiver
from
wild
mammal
studies.
The
waiver
request
was
based
on
the
rationale
that
P.
flocculosa
is
a
naturallyoccurring
soil
microorganism
whose
level
in
the
environment
will
not
significantly
increase
with
greenhouse
use
of
SPORODEX
L
and
that
an
extensive
literature
search
and
the
studies
submitted
to
address
human
health
issues
yielded
no
reports
or
evidence
of
significant
adverse
effects
in
wild
mammals.

Use
of
SPORODEX
L
in
commercial
greenhouses
are
not
expected
to
result
in
increased
exposure
or
adverse
effects
in
wild
mammals.
Wild
mammals
will
not
be
directly
exposed
to
the
product
at
the
time
of
application.
Greenhouse
practices
designed
to
limit
exposure
to
the
outside
environment
will
limit
post­
application
exposure
to
wild
mammals.
Consequently,
testing
is
considered
unnecessary
to
assess
the
risks
of
SPORODEX
L
to
wild
mammals.

6.3
Fish
6.3.1
Freshwater
fish
and
Estuarine/
Marine
animals
(OPPTS
885.4200
and
OPPTS
885.4280,
MRID#
451152­
12)

Plant
Products
Co.
Ltd.
submitted
an
acceptable
justification
for
a
data
waiver
from
freshwater
fish
and
estuarine/
marine
aniimal
studies.
The
waiver
request
was
based
on
the
rationale
that
P.
flocculosa
is
a
naturally­
occurring
microorganism
whose
level
in
the
environment
will
not
significantly
increase
with
greenhouse
use
of
SPORODEX
L
and
that
an
extensive
literature
search
yielded
no
reports
of
adverse
effects
in
freshwater
fish
and
estuarine/
marine
animals.

Use
of
SPORODEX
L
in
commercial
greenhouses
is
not
expected
to
result
in
increased
exposure
or
adverse
effects
in
freshwater
fish
and
estuarine/
marine
animals.
Consequently,
testing
is
considered
unnecessary
to
assess
the
risks
of
SPORODEX
L
to
freshwater
fish
and
estuarine/
marine
animals.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
27
6.4
Arthropods
6.4.1
Terrestrial
arthropods
(OPPTS
885.4340
and
885.4380,
MRID#
451152­
12)

Plant
Products
Co.
Ltd.
submitted
an
acceptable
justification
for
a
data
waiver
from
terrestrial
arthropod
and
honey
bee
testing.
The
waiver
request
was
based
on
the
rationale
that
P.
flocculosa
is
a
naturally­
occurring
microorganism
whose
level
in
the
environment
will
not
significantly
increase
with
greenhouse
use
of
SPORODEX
L
and
that
an
extensive
literature
search
yielded
no
reports
of
adverse
effects
in
terrestrial
arthropods.

Terrestrial
arthropods,
outside
of
greenhouse
facilities
using
SPORODEX
L,
will
not
be
directly
exposed
to
the
product
at
the
time
of
application.
Greenhouse
practices
designed
to
limit
exposure
to
the
outside
environment
will
limit
post­
application
exposure
to
terrestrial
arthropods.
Use
of
SPORODEX
L
in
commercial
greenhouses
is
not
expected
to
result
in
increased
exposure
or
adverse
effects
in
terrestrial
arthropods
and
honeybees.
Consequently,
testing
is
considered
unnecessary
to
assess
the
risks
of
SPORODEX
L
to
terrestrial
arthropods
and
honey
bees.

6.4.2
Aquatic
arthropods
(OPPTS
885.4240,
MRID#
451152.12)

Plant
Products
Co.
Ltd.
submitted
an
acceptable
justification
for
a
data
waiver
from
aquatic
arthropod
studies.
The
waiver
request
was
based
on
the
rationale
that
P.
flocculosa
is
a
naturally­
occurring
microorganism
whose
level
in
the
environment
will
not
significantly
increase
with
greenhouse
use
of
SPORODEX
L
and
that
an
extensive
literature
search
yielded
no
reports
of
adverse
effects
in
aquatic
arthropods.

Use
of
SPORODEX
L
in
commercial
greenhouses
is
not
expected
to
result
in
increased
exposure
or
adverse
effects
in
aquatic
arthropods.
Consequently,
testing
is
considered
unnecessary
to
assess
the
risks
of
SPORODEX
L
to
aquatic
arthropods.

6.5
Non­
arthropod
invertebrates
(OPPTS
885.4240
and
885.4340,
MRID#
451152­
12)

Plant
Products
Co.
Ltd.
submitted
an
acceptable
justification
for
a
data
waiver
from
nonarthropod
invertebrate
studies.
The
waiver
request
was
based
on
the
rationale
that
P.
flocculosa
is
a
naturally­
occurring
microorganism
whose
level
in
the
environment
will
not
significantly
increase
with
greenhouse
use
of
SPORODEX
L
and
that
an
extensive
literature
search
yielded
no
reports
of
adverse
effects
in
non­
arthropod
invertebrates.

Greenhouse
practices
designed
to
limit
exposure
to
the
outside
environment
will
limit
postapplication
exposure
to
non­
arthropod
invertebrates.
Use
of
SPORODEX
L
in
commercial
greenhouses
is
not
expected
to
result
in
increased
exposure
or
adverse
effects
in
non­
arthropod
invertebrates.
Consequently,
testing
is
considered
unnecessary
to
assess
the
risks
of
SPORODEX
L
to
non­
arthropod
invertebrates.

6.6
Microorganisms
(OPPTS
885.1100,
MRID#
451152­
12)
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
28
In
place
of
submitting
microorganism
studies,
Plant
Products
Co.
Ltd.
submitted
a
summary
of
the
host
range
and
mode
of
action
for
P.
flocculosa.
This
information
has
been
reviewed
for
Part
M2,
Product
Characterization
and
Analysis.
The
applicant
has
also
based
a
waiver
rationale
on
the
natural
occurrence
and
limited
additional
exposure
which
will
be
expected
due
to
the
proposed
uses
of
SPORODEX
L.

A
potential
exists,
particularly
in
a
greenhouse
environment
where
conditions
are
optimal
for
growth
(e.
g.,
high
relative
humidity,
controlled
temperatures),
for
P.
flocculosa
to
adversely
affect
non­
target
microorganisms.
Based
on
the
natural
occurrence,
mode
of
action
and
limited
host
range
of
P.
flocculosa,
however,
non­
target
microorganism
testing
with
SPORODEX
L
will
not
be
required
to
assess
the
magnitude
of
this
impact.

6.7
Plants
(OPPTS
885.4300,
MRID#
451152­
12)

6.7.1
Aquatic
plants
Plant
Products
Co.
Ltd.
submitted
an
acceptable
justification
for
a
data
waiver
from
aquatic
plant
testing.
The
waiver
request
was
based
on
observations
of
no
adverse
effects
in
greenhouse
trials
on
target
crops,
the
natural
occurrence
of
P.
flocculosa,
the
proposed
use
pattern
and
an
extensive
literature
search
which
yielded
no
reports
of
adverse
effects
in
aquatic
plants.

Use
of
SPORODEX
L
in
commercial
greenhouses
is
not
expected
to
result
in
increased
exposure
or
adverse
effects
in
aquatic
plants.
Adequate
containment
measures,
aimed
at
minimizing
exposure
to
the
outside
environment,
are
in
place.
Consequently,
testing
is
considered
unnecessary
to
assess
the
risks
of
SPORODEX
L
to
aquatic
plants.

6.7.2
Terrestrial
plants
Plant
Products
Co.
Ltd.
submitted
an
acceptable
justification
for
a
data
waiver
from
terrestrial
plant
studies.
The
waiver
request
was
based
on
observations
of
no
adverse
effects
in
greenhouse
trials,
the
natural
occurrence
of
P.
flocculosa,
the
proposed
use
pattern
and
an
extensive
literature
search
which
yielded
no
reports
of
adverse
effects
in
terrestrial
plants.

Despite
the
common
occurrence
of
P.
flocculosa,
no
incidents
of
adverse
effects
on
terrestrial
plants
have
been
noted.
The
strain
of
P.
flocculosa
used
in
SPORODEX
L
was,
in
fact,
isolated
from
the
leaves
of
red
clover
grown
in
Harrow,
Ontario.
Furthermore,
visual
inspections
of
cucumber,
rose
and
tomato
plants,
treated
with
P.
flocculosa
for
numerous
efficacy
trials,
yielded
no
signs
of
phytotoxicity.

SPORODEX
L
will
be
used
in
commercial
greenhouses
only.
Non­
target
terrestrial
plants
will
not
be
directly
exposed
to
the
product
at
the
time
of
application.
Greenhouse
practices
designed
to
limit
exposure
to
the
outside
environment
will
limit
post­
application
exposure
to
non­
target
terrestrial
plants.
Use
of
SPORODEX
L
in
commercial
greenhouses
is
not
expected
to
result
in
increased
exposure
or
adverse
effects
in
terrestrial
plants.
Consequently,
testing
is
considered
unnecessary
to
assess
the
risks
of
SPORODEX
L
to
non­
target
terrestrial
plants.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
29
Table
6.
1
Risks
of
Pseudozyma
flocculosa
strain
PF­
A22
UL
to
non­
target
organisms
Organism
Exposure
Test
Substance
Conclusions
Birds
Oral/
Pulmonary
/
Inhalation
/
Injection
Waiver
rationale
submitted
in
lieu
of
data
The
waiver
rationale
submitted
by
the
company
was
ACCEPTED
based
on
a
limited
potential
for
risk.

Wild
mammals
Acute
Waiver
rationale
submitted
in
lieu
of
data
The
waiver
rationale
submitted
by
the
company
was
ACCEPTED
based
on
a
limited
potential
for
risk.

Freshwater
fish
Acute
Waiver
rationale
submitted
in
lieu
of
data
The
waiver
rationale
submitted
by
the
company
was
ACCEPTED
based
on
a
limited
potential
for
risk.

Arthropods
Acute
Waiver
rationale
submitted
in
lieu
of
data
The
waiver
rationale
submitted
by
the
company
was
ACCEPTED
based
on
a
limited
potential
for
risk.

Non­
arthropod
invertebrates
Acute
Waiver
rationale
submitted
in
lieu
of
data
The
waiver
rationale
submitted
by
the
company
was
ACCEPTED
based
on
a
limited
potential
for
risk.

Microorganisms
Acute
Waiver
rationale
submitted
in
lieu
of
data
Although
non­
target
microorganisms
may
be
at
potential
risk,
the
waiver
rationale
submitted
by
the
company
was
ACCEPTED
based
on
limited
host
range.

Plants
Acute
Waiver
rationale
submitted
in
lieu
of
data
The
waiver
rationale
submitted
by
the
company
was
ACCEPTED
based
on
a
limited
potential
for
risk.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
30
6.8
Integrated
environmental
toxicology
summary
Acceptable
waiver
rationales
were
submitted
to
address
environmental
toxicology
requirements.
These
waivers
were
based
on
minimal
increased
exposure
of
non­
target
organisms
resulting
from
greenhouse
use
of
SPORODEX
L.
No
reports
of
adverse
effects
on
birds,
wild
mammals,
freshwater
fish,
aquatic
and
terrestrial
arthropods,
non­
arthropod
invertebrates,
and
aquatic
and
terrestrial
plants
organisms
have
been
reported
in
the
literature.
Studies
to
assess
the
effect
of
P.
flocculosa
on
these
organisms
are
not
required.

Pseudozyma
flocculosa
is
a
saprophytic
fungal
epiphyte
and
a
hyperparasite
of
powdery
mildew.
Rapid
death
and
collapse
of
susceptible
host
cells
occurs
via
the
secretion
of
three
fungitoxic
unsaturated
C­
17
fatty
acids
and
an
acyclic
norterpene.
The
fungitoxins
disrupt
susceptible
plasma
membranes
and
cytoplasmic
organelles
while
the
acyclic
norterpene
has
limited
antifungal
potential.
Despite
the
mode
of
action
associated
with
P.
flocculosa,
its
host
range
is
limited
to
mainly
powdery
mildews
(e.
g.,
Sphaerotheca
pannosa
var.
rosae,
S.
fulginea,
Erysiphe
graminis
var
tritici
and
E.
polygoni).
Although
in
vitro
bioassays
have
shown
that
soil­
borne
fungi
such
as
Trichoderma,
Fusarium,
Pythium,
Phytophthora
and
Rhizoctonia
species
and
selected
Gramnegative
(e.
g.,
Xanthomonas
campestris)
and
Gram­
positive
(e.
g.,
Bacillus
subtilis)
bacteria
were
weakly
to
moderately
susceptible
to
P.
flocculosa,
P.
flocculosa
being
a
phyllosphere
epiphyte
and
a
non­
rhizosphere
competent
organism
is
not
expected
to
have
significant
effects
on
soil­
borne
microorganisms.
Based
on
the
limited
host
range
of
P.
flocculosa,
no
non­
target
microorganism
testing
will
be
required.

The
formulants
in
the
end­
use
product
do
not
pose
an
environmental
risk
when
used
at
the
proposed
concentrations
and
application
rate
for
control
of
powdery
mildew
on
roses
and
cucumbers
grown
in
greenhouses.

Chapter
7
Efficacy
data
and
information
A
summary
of
the
efficacy
data
and
information
are
provided
below.

7.1
Effectiveness
7.1.1
Intended
use
For
control
of
powdery
mildew
in
greenhouse
crops:
cut
roses
or
cucumbers
or
potted
roses.
Mix
500
mL
of
SPORODEX
L
for
each
100
L
water
(or
64
U.
S.
fl
oz
per
100
U.
S.
gallons
of
water)
(equivalent
to
approximately
10
5
to
10
6
CFU/
mL).
Add
a
wetting
agent
at
0.
02%.

Apply
up
to
1500
L
of
water
per
ha
(or
150
U.
S.
gallons
of
spray
mixture
per
acre)
for
cut
roses
or
cucumbers
and
1000
L
per
ha
for
potted
roses
(or
100
U.
S.
gallons
per
acre)
.
Spray
foliage
of
plants
to
runoff
at
weekly
intervals,
beginning
when
environmental
conditions
favour
development
of
powdery
mildew
or
at
first
sign
of
the
disease.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
31
Maintain
RH
above
70%
for
12
hours
after
application.

Use
of
chemical
fungicides
at
the
same
time
as
SPORODEX
may
inhibit
this
product's
activity
against
powdery
mildew.

Keep
frozen
at
­20
o
C
or
less
in
the
freezer
until
use;
thaw
at
room
temperature
prior
to
using.

7.1.2
Mode
of
action
SPORODEX
L
is
a
liquid
formulation
containing
Pseudozyma
flocculosa
(synonym
Sporothrix
flocculosa)
at3x10
8
CFU/
mL.
The
strain
of
P.
flocculosa
that
is
the
basis
of
this
product
was
isolated
from
powdery
mildew
on
weeds
near
Harrow,
Ontario
in
1988.
It
has
been
found
to
be
antagonistic
to
most
species
of
powdery
mildew
pathogens,
and
appears
to
be
common
in
horticultural
and
agricultural
environments
where
powdery
mildews
are
found.
Host
range
may
include
antagonism
to
Sphaerotheca
and
Erysiphe
but
it
is
less
active
on
Trichoderma,
Fusarium,
Pythium,
Phytophthora
and
Rhizoctonia
according
to
in
vitro
bioassays.
Pseudozyma
destroys
the
integrity
of
host
cell
membranes
through
the
action
of
fatty
acid
metabolism,
causing
cell
leakage,
but
does
not
appear
to
colonize
host
hyphae.
Optimum
conditions
for
infection
of
host
fungi
are
26
o
C
and
>
70%
RH.
Pseudozyma
will
colonise
leaves
in
the
absence
of
powdery
mildew
but
undergoes
rapid
reproduction
only
when
the
disease
is
present.

7.1.3
Crops
SPORODEX
L
is
intended
for
use
on
greenhouse
roses
and
cucumbers.

7.1.4
Effectiveness
against
pest
Eleven
trials
with
Pseudozyma
flocculosa
on
cucumber
were
conducted
in
the
Netherlands
and
Canada
in
research
or
commercial
greenhouses.
Plants
were
grown
in
rockwool
according
to
normal
hydroponic
production
practices.
SPORODEX
WP
(two
early
formulations)
or
SPORODEX
L
were
applied
at
intervals
as
per
label
directions
to
plants
which
were
usually
inoculated
with
powdery
mildew
(Sphaerotheca
spp.)
For
comparison,
commercial
fungicide
standards
were
applied
as
needed.
Powdery
mildew
(%
diseased
leaf
area
on
whole
plants)
was
assessed
at
intervals
and
an
area
under
disease
progress
curve
(AUDPC)
for
the
whole
season
was
generated
for
comparison
of
treatments.
Cucumbers
were
harvested
and
graded,
and
total
yield
or
first
class
grade
yield
were
recorded.

In
seven
cucumber
trials,
based
on
total
disease
over
the
season,
SPORODEX
WP
provided
1848
control
which
was
significantly
different
from
the
check
but
less
than
that
provided
by
the
chemical
fungicides
(44­
66%).
In
two
comparative
trials
under
moderate
disease
pressure,
a
newer
formulation
of
SPORODEX
WP
showed
significantly
better
control
(>
56%)
than
the
check
and
than
the
older
formulation
which
was
not
effective.
Yield
of
SPORODEX­
treated
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
32
cucumbers
was
generally
greater
than
the
check
and
slightly
lower
than
chemically
treated
plants.

Rose
powdery
mildew
studies
were
conducted
in
Canadian
and
Columbian
greenhouses.
SPORODEX
WP
was
comparable
in
efficacy
to
various
chemical
fungicides
and
often
resulted
in
better
quality
or
yield
of
roses.
The
product
was
less
effective
where
high
humidity
was
not
maintained.

One
confirmatory
trial
with
the
proposed
SPORODEX
L
formulation
showed
that
it
is
as
effective
as
myclobutanil
against
powdery
mildew
on
cucumber.
In
this
trial,
an
application
of
pine
oil
(fertiliser)
in
midseason
adversely
affected
SPORODEX
L
which
resulted
in
lack
of
disease
control
on
lower
leaves
and
showed
that
this
product
is
not
compatible.
Results
are
available
from
two
additional
trials
in
the
Netherlands
and
B.
C.,
which
showed
that
reduction
of
powdery
mildew
and
improved
rose
yield
with
SPORODEX
L
were
comparable
to
results
with
dodemorph­
acetate.
Efficacy
studies
showed
a
need
to
maintain
high
humidity
(>
70%
RH)
for
continued
viability
and
effectiveness
of
SPORODEX
products.

These
studies
show
that
SPORODEX
can
provide
comparable
efficacy
to
chemical
fungicide
sprays
in
controlling
powdery
mildew
and
improving
yield
of
cucumbers
and
quality
of
greenhouse
roses.
SPORODEX
significantly
reduced
powdery
mildew
compared
with
untreated
checks.
Although
the
early
formulation
was
not
as
effective
as
chemical
standards,
limited
trials
with
more
recent
formulations
suggest
that
SPORODEX
L
will
be
as
effective
as
chemical
standards
provided
that
high
humidity
is
maintained.
Further,
it
does
not
cause
phytotoxic
effects
which
indirectly
lowered
yields
as
were
seen
with
some
chemical
treatments.

The
proposed
rate
of
SPORODEX
L
was
500
mL
product
in
100
L
water,
applied
to
runoff
(1500
L/
ha
for
cut
roses
and
cucumbers
and
1000
L/
ha
for
potted
roses).
This
delivers
approximately
1
x
10
9
CFU/
L
of
P.
flocculosa
which
was
the
concentration
used
in
most
of
the
efficacy
trials
with
various
formulations.
A
lower
rate
may
also
be
effective
but
should
not
be
considered
until
crop
management
practices
have
developed
to
give
more
consistent
disease
control
performance
at
the
current
rate.

7.1.5
Total
spray
volume
Mix
500
mL
of
SPORODEX
L
for
each
100
L
water
(or
64
U.
S.
fl
oz
per
100
U.
S.
gallons
of
water)
(equivalent
to
approximately
10
5
to
10
6
CFU/
mL).
Add
a
wetting
agent
at
0.
02%.

Apply
up
to
1500
L
of
water
per
ha
(or
150
U.
S.
gallons
of
spray
mixture
per
acre)
for
cut
roses
or
cucumbers
and
1000
L
per
ha
for
potted
roses
(or
100
U.
S.
gallons
per
acre)
.
Spray
foliage
of
plants
to
runoff
at
weekly
intervals,
beginning
when
environmental
conditions
favour
development
of
powdery
mildew
or
at
first
sign
of
the
disease.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
33
7.2
Phytotoxicity
to
target
plants
(including
different
cultivars),
or
to
target
plant
products
(OECD
7.
4)

No
adverse
effects
of
SPORODEX
formulations
were
noted
in
greenhouse
trials
with
cucumber
or
rose.
The
additive
paraffin
oil
(1%)
used
with
SPORODEX
was
noted
to
cause
a
slight
oedema
(water
blisters)
on
the
underside
of
rose
leaves
of
one
cultivar
and
the
use
of
oil
was
discontinued
in
that
trial.
The
oil
was
typically
used
at
lower
concentrations
in
other
trials
and
is
not
recommended
on
the
SPORODEX
L
label.

7.3
Observations
on
undesirable
or
unintended
side
effects
e.
g.
on
beneficial
and
other
non­
target
organisms,
on
succeeding
crops,
other
plants
or
parts
of
treated
plants
used
for
propagating
purposes
(e.
g.
seed,
cutting,
runners)
(OECD
7.5)

SPORODEX
L
was
tested
in
commercial
greenhouses
throughout
its
development,
using
typical
production
practices
including
IPM
and
biological
control
organisms.
In
these
efficacy
trials,
observations
suggested
that
the
product
has
no
adverse
effect
on
crop
plants,
or
on
beneficial
insects
or
mites
with
respect
to
pest
control.
However,
direct
assays
to
confirm
no
adverse
effect
of
SPORODEX
L
on
specific
biocontrol
insects
and
arthropods
were
not
conducted.

7.3.1
Impact
on
succeeding
crops
(OECD
7.
5.
1)

Not
applicable
to
greenhouse
use.

7.3.2
Impact
on
adjacent
crops
(OECD
7.
5.
2)

Not
applicable
to
greenhouse
use;
adjacent
crops
(if
any)
are
typically
grown
within
separate
compartments.

7.3.3
Impact
on
seed
viability
(OECD
7.5.3)

Not
applicable
to
proposed
crops.

7.4
Economics
According
to
the
applicant,
the
farm
gate
value
of
greenhouse
cucumbers
in
Ontario
is
$25
million.
There
are
also
greenhouse
areas
in
BC,
Alberta
and
Quebec.
Crops
are
worth
$100­
200
per
ha
annually.
There
are
about
24
ha
of
greenhouse
roses
in
Canada,
mostly
in
Ontario,
with
some
operations
in
BC,
Alberta
and
Quebec.
Grade
#1
roses
are
priced
at
$0.50
per
stem.

Although
it
rarely
affects
fruit,
powdery
mildew
spreads
rapidly
on
leaves
and
can
cause
a
loss
of
photosynthetic
area
and
water
stress,
leading
to
reduced
flower
production
or
yield
and
up
to
100%
loss.
Mildew
can
also
affect
marketability
and
price
of
roses
as
there
is
zero
tolerance
for
the
presence
of
white
mildew
spots
on
the
leaves
and
blooms
and
they
will
be
downgraded.
The
price
difference
to
growers
for
grade
#1
to
grade
#2
roses
is
$0.15
per
stem,
and
control
of
mildew
could
potentially
increase
revenues
by
$6,
000
per
ha.
Fungicides
are
currently
used
to
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
34
control
mildew
but
can
have
adverse
effects
on
yield,
fruit
and
flower
quality.

7.5
Sustainability
7.5.1
Survey
of
alternatives
Powdery
mildew
is
currently
managed
by
environment
control,
tolerant
cultivars,
sanitation
and
fungicides.
The
trend
in
greenhouse
production
is
to
reduce
chemical
pesticide
use
as
much
as
possible.
Thus,
there
is
a
need
for
alternative
products
to
use
in
the
disease
management
program.

7.5.1.1
Non­
chemical
control
practices
Powdery
mildew
is
partly
managed
by
environment
control;
however
this
is
difficult
to
balance
because
the
different
stages
of
disease
development
are
favoured
by
different
conditions.
For
example,
both
low
humidity
and
free
water
on
leaves
followed
by
rapid
drying
have
been
found
to
reduce
disease,
yet
daily
fluctuations
in
humidity
can
increase
disease.
In
general,
growers
should
avoid
conditions
which
lead
to
succulent
foliage,
ie.
shading,
overcrowding,
overwatering
or
overfertilizing.
The
fungal
spores
will
not
survive
long
outside
of
host
plant
material,
so
a
thorough
cleanup
and
break
period
of
2­
3
weeks
between
crops
can
reduce
carryover
of
inoculum.
Seedlings
which
are
already
started
should
be
cultivated
in
isolation
from
the
older
producing
plants.
Teardown
and
replant
of
the
cucumber
crop
is
a
labour
intensive
operation
due
to
the
volume
of
vine
material
handled,
so
the
plants
are
usually
maintained
as
long
as
is
profitable.

Mildew­
resistant
cucumber
and
rose
cultivars
are
not
available
in
practice
for
Canadian
conditions;
although
some
tolerance
is
available,
these
cultivars
may
not
be
commercially
desirable.
For
instance,
in
B.
C.
cucumber
production,
more
resistant
varieties
are
grown
in
fall
when
mildew
pressure
is
high;
however,
they
are
lower
yielding
and
more
prone
to
Botrytis
and
gummy
stem
than
mildew­
susceptible
varieties
grown
earlier
in
the
year.
Choice
of
rose
varieties
is
dependant
on
market
acceptability
for
colour
and
other
characteristics
rather
than
tolerance
to
powdery
mildew.

7.5.1.2
Chemical
Control
Practices
Few
chemical
products
are
available
for
powdery
mildew
control
in
greenhouses.
Benomyl,
myclobutanil,
and
sulfur
are
registered
in
Canada
for
cucumber
and
dodemorph­
acetate
and
copper
are
registered
for
roses.
The
most
effective
products
are
systemic,
must
be
applied
frequently,
may
be
toxic
to
beneficials
and
are
prone
to
development
of
resistance
in
the
powdery
mildew
pathogens.
Both
myclobutanil
and
dodemorph
were
noted
to
cause
phytotoxicity
to
flowers
and
fruit
under
some
conditions
and
reduced
leaf
size
has
been
reported
for
both
cucumber
and
rose.
Benomyl
is
registered
but
not
expected
to
be
marketed
beyond
2001.
Silicon
has
been
investigated
as
a
disease
preventative,
either
applied
into
hydroponic
solution
or
as
a
foliage
spray,
but
has
not
been
consistently
effective
on
its
own.
Milsana
is
another
non­
fungicidal
product
under
investigation
but
not
used
commercially
in
Canada.
The
trend
in
greenhouse
production
is
to
reduce
chemical
pesticide
use
as
much
as
possible.
Thus,
there
is
a
need
for
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
35
alternative
products
to
use
in
the
disease
management
program.

Table
7.
5­
1
Alternative
disease
control
products
Active
Ingredient
End­
Use
Products
Fungicide
Activity
Site
Application
Rate
(product/
1000
L)
Comments
Cucumber
Rose
Benomyl
Benlate
50WP
(+
Manzate)
tubulin
(multisite)
550­
850
g
2.25
kg
No
longer
marketed
prone
to
resistance
Myclobutanil
Nova
40W
demethylation
340
g/
ha
Can
affect
leaf
and
fruit
growth,
prone
to
resistance
Sulfur
Kumulus,
MicroNiasul
multisite
1.
2­
1.
5
kg
Harmful
to
some
beneficial
mites,
can
be
phytotoxic
Copper
Phyton
27
multisite
1.
25­
2.
5
L
Dodemorph
acetate
Meltatox
40EC
isomerase
2.
5
L
Can
reduce
bloom
quality
7.5.2
Compatibility
with
current
management
practices
including
IPM
SPORODEX
L
has
potential
to
reduce
or
replace
chemical
fungicide
sprays
on
cucumbers
and
roses
and
efficacy
trials
showed
that
it
can
be
alternated
with
some
of
these
products.
SPORODEX
L
also
appears
to
be
compatible
with
IPM
practices
for
control
of
insects
and
mites
(see
section
7.
3).
However,
SPORODEX
L
has
not
been
tested
for
compatibility
with
all
chemical
products
or
with
other
microbial
disease
control
organisms;
therefore
the
grower
should
be
referred
to
the
manufacturer
for
updated
information.

IPM
practices
currently
include
the
monitoring
of
crop
for
signs
of
early
disease
which
is
necessary
to
ensure
that
SPORODEX
L
is
applied
at
the
earliest
opportunity
for
maximum
effectiveness.
At
present,
the
value
of
SPORODEX
L
is
limited
by
its
susceptibility
to
changing
environmental
conditions.
Growers
and
extension
staff
will
need
to
invest
further
work
in
determining
the
best
local
production
practices
for
viability
and
efficacy
of
SPORODEX
L
in
the
greenhouse
to
obtain
optimum
powdery
mildew
control
and
thereby
reduce
the
need
for
chemical
products.

7.5.3
Contribution
to
risk
reduction
It
is
expected
that
SPORODEX
L
will
be
used
in
greenhouses
to
control
powdery
mildew
in
situations
of
lower
disease
pressure
and
at
the
beginning
of
the
growing
season
to
delay
the
progress
of
the
disease.
In
this
way
it
may
aleviate
or
defer
the
need
for
chemical
fungicide
applications
thus
reducing
the
associated
risks
of
pesticide
resistance,
effects
to
workers
and
to
the
environment.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
36
7.5.4
Information
on
the
occurrence
or
possible
occurrence
of
the
development
of
resistance
Powdery
mildew
pathogens
have
been
known
to
develop
resistance
to
chemical
fungicides;
however,
resistance
to
Pseudozyma
flocculosa
is
less
likely
because
of
its
broad
mode
of
action
and
lack
of
persistence
on
the
crop
plant.
Pseudozyma
flocculosa
destroys
the
integrity
of
host
cell
membranes,
causing
cell
leakage.
Optimum
conditions
for
colonization
do
not
occur
continuously
in
the
greenhouse
environment.
It
will
colonise
leaves
in
the
absence
of
powdery
mildew
but
undergoes
rapid
reproduction
only
when
the
disease
is
present.
For
these
reasons,
it
is
not
anticipated
that
resistance
to
P.
flocculosa
will
develop;
however,
as
a
general
principal,
it
is
best
not
to
rely
continuously
on
any
one
product
for
disease
control.

SPORODEX
L
does
have
a
role
in
prolonging
effectiveness
of
chemical
fungicides.
By
reducing
pathogen
populations
and
the
number
of
fungicide
sprays
applied
for
control
of
powdery
mildew,
SPORODEX
L
may
reduce
pressure
on
the
pathogen
to
develop
resistance
to
more
site­
specific
fungicides.

7.6
Conclusions
7.6.1
Summary
SPORODEX
L
is
a
liquid
formulation
containing
Pseudozyma
flocculosa
at
3
x
10
8
CFU/
mL
for
control
of
powdery
mildew
in
greenhouse
roses
and
cucumbers.
The
proposed
rate
of
SPORODEX
L
is
500
mL
product
in
100
L
water
(or
64
U.
S.
fl
oz
per
100
U.
S.
gallons
of
water),
applied
to
runoff
(1500
L/
ha
for
cut
roses
and
cucumbers
(or
150
U.
S.
gallons
of
spray
mixture
per
acre)
and
1000
L/
ha
for
potted
roses
(or
100
U.
S.
gallons
per
acre)).

Eleven
trials
with
Pseudozyma
flocculosa
on
cucumber
were
conducted
in
the
Netherlands
and
Canada
in
research
or
commercial
greenhouses.
Five
rose
powdery
mildew
studies
were
conducted
in
Canadian
and
Columbian
greenhouses.
SPORODEX
significantly
reduced
powdery
mildew
compared
with
untreated
checks.
Although
the
early
formulation
was
not
as
effective
as
chemical
standards,
limited
trials
with
more
recent
formulations
suggest
that
SPORODEX
L
will
be
as
effective
as
chemical
standards
provided
that
high
humidity
is
maintained.
Further,
it
does
not
cause
phytotoxic
effects
which
indirectly
lowered
yields
as
were
seen
with
some
chemical
treatments.
Further
work
is
needed
on
managing
the
greenhouse
environment
for
full
disease
control
benefits
of
SPORODEX
L
to
be
realized.

SPORODEX
L
is
a
microbial
product
which
may
be
affected
by
co­
application
of
fungicides
and
other
products.
The
label
precautions
should
be
expanded
to
advise
the
grower
of
this
and
include
guidance
for
better
performance.
SPORODEX
L
has
not
been
shown
to
adversely
affect
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
37
other
tools
such
as
biocontrol
agents,
shows
no
phytotoxic
effects
on
the
crop
and
is
generally
compatible
with
IPM
practices
which
are
being
adopted
for
greenhouse
production.

Table
7.
6­
1
Summary
of
label
proposals
and
recommendations
Proposed
Recommendation
(based
on
Value
Assessment)

Greenhouse
crops
Cucumber
as
proposed
Roses
(potted
or
cut)
as
proposed
Rate
500
mL
/100L
of
water
(or
64
U.
S.
fl
oz
per
100
U.
S.
gallons
of
water)
with
20
mL
wetting
agent
(3
U.
S.
fl
oz).
Use
up
to
1500
L
spray
per
ha
for
cut
roses,
cucumbers
(or
150
U.
S.
gallons
of
spray
mixture
per
acre)
,
up
to
1000
L
for
potted
roses
(or
100
U.
S.
gallons
per
acre)
as
proposed
Application
method
Diluted
spray
applied
to
foliage
to
runoff
as
proposed
Timing
of
applications
Weekly
from
first
disease
or
when
environmental
conditions
favour
development
of
disease
as
proposed
Conditions
Maintain
RH
>70%
for
12
hours
Donotapplyatsame
time
as
chemical
fungicides
provide
additional
details/
guidance
Chapter
8
Overall
risk
assessment
conclusions
8.1
Product
characterization
and
analysis
The
product
characterization
data
for
both
Pseudozyma
flocculosa
strain
PF­
A22
UL
and
SPORODEX
L
are
adequate
to
assess
their
safety
to
human
health.
The
technical
material
was
fully
characterized
and
the
specifications
were
supported
by
the
analysis
of
a
sufficient
number
of
batches.
Quality
control
procedures
employed
during
product
manufacture
and
formulation
are
adequate
to
ensure
an
absence
of
contaminating
microorganisms
of
concern
including
primary
human
and
animal
pathogens.
However,
due
to
the
microbial
contamination
noted
in
the
representative
quality
control
data,
the
submission
of
certificates
of
analysis
will
be
required
for
all
production
batches
of
SPORODEX
L
as
a
condition
of
registration
by
the
PMRA
and
the
EPA.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
38
Additional
storage
stability
data
on
pilot­
scale
or
production­
scale
batches
are
required
to
ensure
product
performance
and
safety.
Until
these
data
are
available,
an
expiration
date
of
3
months
when
the
product
is
stored
at
­20

C
is
required
on
the
product
labels.

8.2
Toxicity
and
infectivity
The
acute
toxicity
and
infectivity
studies
submitted
in
support
of
P.
flocculosa
and
of
SPORODEX
L
were
determined
to
be
sufficiently
complete
to
permit
a
decision
on
registration.
Pseudozyma
flocculosa
was
of
low
toxicity
in
the
rat
when
administered
via
the
oral
and
dermal
routes.
Slight
toxicity
was
observed
when
P.
flocculosa
was
administered
via
the
pulmonary
and
intraperitoneal
routes.
Pseudozyma
flocculosa
was
not
pathogenic
or
infective
via
the
oral
and
intraperitoneal
routes.
Pathogenicity
via
the
pulmonary
route
could
not
be
determined.
No
dermal
irritation
was
observed
but
mild
ocular
irritation
was
noted.
Waiver
requests
were
submitted
to
address
the
use
of
alternative
test
substances
(e.
g.,
SPORODEX
WP,
MPCA
produced
by
the
test
facility)
in
the
acute
toxicity
and
infectivity
studies.
These
waivers
were
based
on
the
nature
of
the
formulation
ingredients
in
SPORODEX
L
and
the
reduced
irritation
potential
of
a
liquid
formulation
compared
to
that
of
a
wettable
powder
formulation.

8.3
Exposure
The
potential
for
dermal,
eye
and
inhalation
exposure
for
pesticide
handlers
exists,
with
the
major
source
of
exposure
to
workers
being
generally
dermal.
To
mitigate
dermal
and
inhalation
exposure
and
risk
to
workers
during
mixing/
loading,
application
and
post­
application
activities,
use
of
appropriate
Personal
Protective
Equipment
(PPE)
will
be
required.
PPE
will
include
longsleeved
shirts,
long
pants,
waterproof
gloves,
dust/
filter
respirator,
shoes
and
socks.
Furthermore
a
Restricted­
Entry
Interval
(REI)
of
4
hours
is
required
for
early
entry
(post­
application)
workers
or
other
persons
entering
treated
greenhouses.

It
is
assumed
that
most
microorganisms
contain
substances
that
would
elicit
positive
hypersensitivity
reactions.
Pseudozyma
flocculosa
strain
PF­
A22
UL
is
considered
a
potential
sensitizing
agent,
and
a
"POTENTIAL
SENSITIZER"
statement
will
be
required
on
the
principal
display
panel
of
the
TGAI
and
end­
use
formulation
labels.

8.4
Food
and
feed
residues
Although
Pseudozyma
species
are
ubiquitous
in
nature
and
have
been
isolated
from
a
wide
variety
of
plant
surfaces
including
leaf
litter,
clover,
maize
and
cucumber,
no
adverse
effects
from
dietary
exposure
have
been
attributed
to
natural
populations
of
Pseudozyma
flocculosa.
Residues
of
the
active
micoorganism
can
be
easily
removed
from
treated
commodities
by
washing,
cooking,
peeling
and
processsing.
Even
if
residues
are
not
removed,
dietary
exposure
to
the
microbial
agent
is
unlikely
to
result
in
any
undue
hazard
to
consumers
because
no
adverse
effects
were
observed
at
maximum
hazard
dose
levels
in
the
submitted
Tier
I
acute
oral
study.
Furthermore,
an
extensive
literature
search
yielded
no
reports
of
mammalian
toxins
being
produced
by
P.
flocculosa.
Based
on
the
low
level
of
toxicity
and
lack
of
pathogenic
potential
for
the
active
microorganism
observed
in
the
Tier
I
acute
oral
study,
there
are
no
acute,
subchronic
or
chronic
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
39
toxicological
concerns
for
P.
flocculosa.
Therefore,
the
establishment
of
a
a
tolerance
for
residues
of
P.
flocculosa
strain
PF­
A22
UL
is
not
required
(rather
an
exemption
from
the
requirement
of
a
tolerance
will
be
granted)
as
defined
under
Section
408
of
the
Federal
Food
Drug
and
Cosmetic
Act.

8.5
Environmental
assessment
Acceptable
waivers
were
submitted
to
address
environmental
toxicology
requirements.
Nontarget
organisms,
including
birds,
wild
mammals,
freshwater
fish,
aquatic
and
terrestrial
arthropods,
non­
arthropod
invertebrates,
and
aquatic
and
terrestrial
plants
are
not
expected
to
face
increased
exposure
to
P.
flocculosa
due
to
use
of
SPORODEX
L
in
commercial
greenhouses.
No
reports
of
adverse
effects
on
these
non­
target
organisms
have
been
reported
in
the
literature.
Studies
to
assess
the
effect
of
P.
flocculosa
on
these
organisms
are
not
required.

Pseudozyma
flocculosa
is
a
saprophytic
fungal
epiphyte
and
a
hyperparasite
of
powdery
mildew.
Rapid
death
and
collapse
of
susceptible
host
cells
occurs
via
the
secretion
of
three
fungitoxic
unsaturated
C­
17
fatty
acids
and
an
acyclic
norterpene.
The
fungitoxins
disrupt
susceptible
plasma
membranes
and
cytoplasmic
organelles
while
the
acyclic
norterpene
has
limited
antifungal
potential.
Despite
the
mode
of
action
associated
with
P.
flocculosa,
its
host
range
is
limited
to
mainly
powdery
mildews
(e.
g.,
Sphaerotheca
pannosa
var.
rosae,
S.
fulginea,
Erysiphe
graminis
var
tritici
and
E.
polygoni).
Although
in
vitro
bioassays
have
shown
that
soil­
borne
fungi
such
as
Trichoderma,
Fusarium,
Pythium,
Phytophthora
and
Rhizoctonia
species
and
selected
Gramnegative
(e.
g.,
Xanthomonas
campestris)
and
Gram­
positive
(e.
g.,
Bacillus
subtilis)
bacteria
were
weakly
to
moderately
susceptible
to
P.
flocculosa,
P.
flocculosa
being
a
phyllosphere
epiphyte
and
a
non­
rhizosphere
competent
organism
is
not
expected
to
have
significant
effects
on
soilborne
microorganisms.
Based
on
the
limited
host
range
of
P.
flocculosa,
no
non­
target
microorganism
testing
will
be
required.

The
formulants
in
the
end­
products,
SPORODEX
L,
do
not
pose
an
environmental
risk
when
used
at
the
proposed
concentrations
and
application
rate
for
control
of
powdery
mildew
on
roses
and
cucumbers
grown
in
greenhouses.
Consequently,
SPORODEX
L
is
expected
to
pose
little
environmental
risk
when
used
in
accordance
with
the
label
directions.
Furthermore,
no
special
precautionary
or
environmental
hazard
statement
is
required
on
the
label
for
SPORODEX
L.

8.
6
Efficacy
assessment
SPORODEX
L
applied
at
proposed
label
rates
can
be
effective
in
controlling
powdery
mildew
on
greenhouse
cucumber
and
roses
provided
that
high
humidity
is
maintained.
Further
work
is
needed
on
managing
the
greenhouse
environment
for
full
disease
control
benefits
of
SPORODEX
L
to
be
realized.
SPORODEX
L
has
not
been
shown
to
adversely
affect
other
tools
such
as
biocontrol
agents,
shows
no
phytotoxic
effects
on
the
crop
and
is
generally
compatible
with
IPM
practices
which
are
being
adopted
for
greenhouse
production.

Chapter
9
Risk
Management
Considerations
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
40
9.1
Public
interest
finding
The
Agency
believes
the
use
of
Pseudozyma
flocculosa
strain
PF­
A22
UL
under
this
conditional
registration
would
be
in
the
public
interest.
The
criteria
for
Agency
evaluation
of
public
interest
findings
is
outlined
in
51
CFR
No.
43,
Wednesday
March
5,
1986.
Under
part
IV.
A.,
the
proposed
product
may
qualify
for
an
automatic
presumptive
finding
if
it
is
for
minor
use,
is
a
unique
replacement
for
pesticides
of
concern,
or
is
for
use
against
a
public
health
pest.

Pseudozyma
flocculosa
strain
PF­
A22
UL
is
intended
for
formulation
into
end­
use
products
for
control
of
powdery
mildew
on
greenhouse­
grown
cucumbers
and
roses.
These
uses
are
for
minor
use
crops
and,
therefore,
the
product
qualifies
for
an
automatic
presumptive
finding
and
its
use
is
presumed
to
be
in
the
public
interest.

9.2
Determination
of
3(
c)(
7)(
C)
eligibility
Pursuant
to
FIFRA
section
3(
c)(
7)(
C),
EPA
may
conditionally
register
a
new
pesticide
active
ingredient
if:
1)
insufficient
time
has
elapsed
since
the
imposition
of
the
data
requirement
for
those
data
to
be
developed
and
all
other
required
data
have
been
submitted,
2)
the
use
of
the
pesticide
product
during
the
period
of
the
conditional
registration
will
not
cause
any
unreasonable
adverse
effect
on
human
health
and
the
environment,
and
3)
the
registration
and
the
use
of
the
pesticide
during
the
conditional
registration
is
in
the
public
interest.
The
Agency
believes
that
all
these
criteria
have
been
fulfilled.

For
Pseudozyma
flocculosa
strain
PF­
A22
UL
and
the
end­
product,
SPORODEX
L,
the
first
criterion
under
FIFRA
section
3(
c)(
7)(
C)
mentioned
above
has
been
met.
Insufficient
time
has
elapsed
since
the
imposition
of
the
following
outstanding
confirmatory
data
requirement:
Acute
Pulmonary
Toxicity/
Pathogenicity.
Agency
scientists
have
reviewed
the
data
submitted
or
cited
by
Plant
Products
Co.
Ltd.
with
respect
to
health
effects
and
ecological
effects
and
have
identified
no
unreasonable
adverse
effects
to
human
health
or
to
non­
target
organisms.
However,
to
confirm
these
expected
results,
the
additional
data
described
below
will
be
required
as
a
condition
of
registration.

A
complete
acute
pulmonary
toxicity
/
pathogenicity
study
(U.
S.
EPA
OPPTS
885.3150)
must
be
conducted
using
the
technical
grade
active
ingredient
(TGAI)
(Pseudozyma
flocculosa
strain
PF­
A22
UL)
and
the
sterile
filtrate
of
the
production
culture.

The
two
acute
pulmonary
studies
that
were
submitted
did
not
have
fully
acceptable
toxicity
and
pathogenicity
results
contained
in
a
single
study.
Both
of
these
studies
were
considered
supplemental.
However,
taken
together,
parts
of
each
study
were
acceptable
for
making
a
regulatory
decision.
That
is,
the
acute
pulmonary
toxicity/
pathogenicity
study
had
acceptable
pathogenicity
data,
but
not
toxicity
data
and
the
acute
pulmonary
toxicity/
pathogenicity
range
finding
study
had
acceptable
toxicity
data,
but
did
not
address
pathogenicity.
In
addition,
the
Agency
decided
that
it
would
be
prudent
to
test
the
sterile
filtrate
of
the
production
batch
to
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
41
determine
whether
there
were
any
toxic
components
of
concern.
Testing
the
sterile
filtrate
would
not
have
been
foreseeable
by
the
registrant
and
a
period
reasonable
sufficient
for
generation
of
the
data
has
not
elapsed.
Thus,
a
confirmatory
acute
pulmonary
toxicity
/
pathogenicity
study
using
the
TGAI
and
testing
of
the
sterile
filtrate
from
the
production
culture
will
be
required
to
provide
this
additional
information
as
a
condition
of
registration.
This
study
must
be
submitted
to
the
EPA
no
later
than
October
3,
2003.

All
other
required
data
for
registration
have
been
submitted.
The
Agency
is
also
imposing
a
continuing
monitoring
requirement
on
the
registrant
as
a
term
of
registration;
this
requirement
is
not
a
3(
c)(
7)(
C)
condition
and
is
not
connected
to
the
two
year
term
of
the
conditional
registration.
In
addition,
the
registrant
may
submit
further
storage
stability
studies
to
support
a
change
in
the
labeling
to
increase
the
labeled
shelf
life
beyond
three
months.
The
submitted
storage
stability
study,
however,
is
sufficient
to
support
the
current
3
month
label
language.

The
applicant
has
submitted
or
cited
data
to
allow
EPA
to
make
the
finding
necessary
to
satisfy
the
second
criterion
for
conditional
registration
under
FIFRA
3(
c)(
7)(
C)
as
mentioned
above.
Plant
Products
Co.
Ltd.
submitted
and/
or
cited
satisfactory
data
pertaining
to
the
proposed
use.
The
human
health
effects
data
and
non­
target
organism
effects
data
are
considered
sufficient
for
the
period
of
the
conditional
registration
(2
years).
These
data
demonstrate
that
no
foreseeable
human
health
hazards
or
ecological
effects
are
likely
to
arise
from
the
use
of
the
product
and,
under
the
terms
and
conditions
of
the
registration.
As
any
potential
inhalation
risk
that
is
raised
by
the
supplementary
acute
pulmonary
toxicity/
pathogenicity
data
is
primarily
a
worker
risk,
EPA
is
requiring
that
a
respirator
be
worn
by
workers
to
limit
any
inhalation
exposures.
In
addition,
a
Restricted­
Entry
Interval
(REI)
of
4
hours
is
required
for
early
entry
(post­
application)
workers
or
other
persons
entering
treated
greenhouses.
Accordingly,
the
Agency
has
concluded
that
use
of
the
pesticide
product
during
the
period
of
the
conditional
registration
will
not
cause
any
unreasonable
adverse
effect
on
human
health
and
the
environment
The
Agency
also
believes
that
the
third
criterion
for
a
FIFRA
3(
c)(
7)(
C)
conditional
registration
has
been
fulfilled
because
the
use
of
Pseudozyma
flocculosa
strain
PF­
A22
UL
under
this
registration
would
be
in
the
public
interest.
The
criteria
for
Agency
evaluation
of
public
interest
findings
is
outlined
in
51
FR
7628
(Mar.
5,
1986).
The
proposed
product
qualifies
for
an
automatic
presumptive
finding
because
all
intended
uses
are
for
minor
use
crops,
i.
e.,
greenhousegrown
cucumbers
and
roses.
In
addition,
the
Agency
is
not
aware
of
any
other
information
which
would
alter
EPA's
presumption
that
use
of
this
pesticide
during
the
period
of
conditional
registration
would
be
in
the
public
interest.

Therefore,
Pseudozyma
flocculosa
strain
PF­
A22
UL
is
eligible
for
a
FIFRA
3(
c)(
7)(
C)
conditional
registration.
The
proposed
registration
sites
are
for
greenhouse­
grown
cucumbers
and
roses.

9.3
Terms
and
conditions
of
registration
The
active
ingredient
Pseudozyma
flocculosa
strain
PF­
A22
UL,
and
the
formulated
product
SPORODEX
L
for
control
of
powdery
mildew
on
greenhouse­
grown
roses
and
cucumbers
have
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
42
been
granted
temporary
registration
pursuant
to
Section
17
of
the
Pest
Control
Products
Regulations
(Canada/
PMRA)
and
a
conditional
registration
under
Section
3(
c)(
7)(
C)
of
the
Federal
Insecticide
Fungicide
and
Rodenticide
Act
(U.
S.
EPA).

This
FIFRA
3(
c)(
7)(
C)
conditional
registration
will
automatically
expire
at
midnight,
September
30,
2004.

The
following
list
gives
the
terms
and
conditions
of
the
registration.

1)
A
complete
acute
pulmonary
toxicity
/
pathogenicity
study
(U.
S.
EPA
OPPTS
885.3150)
must
be
conducted
using
the
technical
grade
active
ingredient
(TGAI)
(Pseudozyma
flocculosa
strain
PF­
A22
UL)
and
the
sterile
filtrate
of
the
production
culture.
This
study
must
be
submitted
to
the
EPA
no
later
than
October
3,
2003.

2)
Certificates
of
analysis
must
be
submitted
to
the
Agency
prior
to
the
release
of
all
production
batches
of
SPORODEX
L
biological
fungicide
(U.
S.
EPA
OPPTS
885­
1100
through
885­
1600).
Bioassay
results,
conidial,
total
aerobic
flora,
enterobacteria,
enterococci,
fecal
coliform,
E.
coli,
Staphylococci
and
Salmonella
counts
must
be
determined
for
each
production
batch
and
be
included
in
each
certificate
of
analysis.
Certificates
of
analysis
must
be
submitted
until
sufficient
consistency
with
regards
to
microbial
contaminants
has
been
established.

3)
Additional
storage
stability
data
(OPPTS
830.6317)
derived
from
at
least
five
production­
scale
or
pilot­
scale
batches
are
required
to
ensure
product
performance
and
safety
during
the
shelf­
life
of
the
product.
SPORODEX
L
Biological
Fungicide
should
be
tested
over
a
period
of
time
and
in
accordance
with
the
desired
storage
and
use
conditions
appearing
on
the
product
label.
An
interim
expiration
date
of
3
months
from
the
date
of
manufacture
when
SPORODEX
L
Biological
Fungicide
is
stored
at
­20

C
is
required
until
additional
data
are
submitted
and
approved
by
the
Agency.
For
consideration
of
these
data
prior
to
the
expiration
date
of
the
conditional
registration,
additional
data
should
be
submitted
to
the
Agency
no
later
than
October
3,
2003.

4)
Finally,
although
a
skin
sensitization
study
on
the
microbial
active
ingredient
Pseudozyma
flocculosa
strain
PF­
A22
UL
was
not
required
by
the
Agency,
the
reporting
of
any
incidents
of
hypersensitivity
subsequent
to
registration
is
a
standard
practice
for
microbial
products.
The
registrant
will
be
expected
to
report
any
subsequent
findings
of
hypersensitivity
or
other
health
incidents
to
workers,
applicators,
or
bystanders
exposed
to
the
MPCA
(microbial
pest
control
agent)
as
an
ongoing
condition
of
registration.
Incident
reports
under
FIFRA
section
6(
a)
2
are
to
include
details
such
as
a
description
of
the
MPCA
and
formulation,
frequency,
duration
and
routes
of
exposure
to
the
material,
clinical
observations,
and
any
other
relevant
information.

9.4
Tolerance
EPA
has
established
an
exemption
from
the
requirement
of
a
tolerance
for
residues
of
the
Pseudozyma
flocculosa
strain
PF­
A22
UL
in
or
on
all
food
commodities.
Based
on
the
toxicology
data
submitted
and
other
relevant
information
in
the
Agency's
files,
there
is
reasonable
certainty
no
harm
will
result
from
aggregate
exposure
of
residues
of
Pseudozyma
flocculosa
strain
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
43
PF­
A22
UL
to
the
U.
S.
population,
including
infants
and
children,
under
reasonably
foreseeable
circumstances
when
the
microbial
pesticide
product
is
used
as
labeled.
This
includes
all
anticipated
dietary
exposures
and
all
other
exposures
for
which
there
is
reliable
information.
The
Agency
has
arrived
at
this
conclusion
based
on
data
submitted
demonstrating
low
toxicity
at
the
maximum
doses
tested
and
a
lack
of
information
showing
adverse
effects
from
exposure
to
naturally
occurring
P.
flocculosa
as
well
as
a
consideration
of
the
product
as
currently
registered
and
labeled.
As
a
result,
EPA
establishes
an
exemption
from
tolerance
requirements
pursuant
to
FFDCA
408(
c)
and
(d)
for
residues
of
Pseudozyma
flocculosa
strain
PF­
A22
UL
in
or
on
all
food
commodities.

FFDCA
section
408
provides
that
EPA
shall
apply
an
additional
tenfold
margin
of
exposure
(safety)
for
infants
and
children
in
the
case
of
threshold
effects
to
account
for
prenatal
and
postnatal
toxicity
and
the
completeness
of
the
data
base
unless
EPA
determines
that
a
different
margin
of
exposure
(safety)
will
be
safe
for
infants
and
children.
Margins
of
exposure
(safety)
are
often
referred
to
as
uncertainty
(safety)
factors.
In
this
instance,
based
on
all
the
available
information,
the
Agency
concludes
that
P.
flocculosa
strain
PF­
A22
UL
is
practically
non­
toxic
to
mammals,
including
infants
and
children.
Thus,
there
are
no
threshold
effects
of
concern
and,
as
a
result
the
provision
requiring
an
additional
margin
of
safety
does
not
apply.
Further,
the
provisions
of
consumption
patterns,
special
susceptibility,
and
cumulative
effects
do
not
apply.
As
a
result,
EPA
has
not
used
a
margin
of
exposure
(safety)
approach
to
assess
the
safety
of
P.
flocculosa
strain
PF­
A22
UL.

9.5
Codex
harmonization
There
are
no
Codex
harmonization
considerations
since
there
are
no
Codex
Maximum
Residue
Limits
set
for
food
use
of
this
active
ingredient.

9.6
Risk
mitigation
There
is
minimal
or
negligible
potential
risk
to
wildlife,
or
ground
and
surface
water
contamination
for
products
containing
this
active
ingredient.
Dietary
risk
will
be
adequately
mitigated
by
washing,
peeling,
cooking
and
processing
of
treated
foods.
To
mitigate
dermal
and
inhalation
exposure
and
risk
to
workers
during
mixing/
loading,
application
and
post­
application
activities,
use
of
appropriate
Personal
Protective
Equipment
(PPE)
will
be
required.
PPE
will
include
long­
sleeved
shirts,
long
pants,
waterproof
gloves,
dust/
filter
respirator,
shoes
and
socks.
Furthermore
a
Restricted­
Entry
Interval
(REI)
of
4
hours
is
required
for
early
entry
postapplication
workers
or
other
persons
entering
treated
greenhouses.

It
is
assumed
that
most
microorganisms
contain
substances
that
would
elicit
positive
hypersensitivity
reactions.
Pseudozyma
flocculosa
strain
PF­
A22
UL
is
considered
a
potential
sensitizing
agent,
and
a
"POTENTIAL
SENSITIZER"
statement
will
be
required
on
the
principal
display
panel
of
the
TGAI
and
end­
use
formulation
labels.

No
special
precautionary
or
environmental
hazard
statements
are
required
on
the
label
for
SPORODEX
L.
There
are
no
reports
of
adverse
effects
due
to
the
use
of
P.
flocculosa.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
44
Exposure
to
P.
flocculosa
due
to
use
of
SPORODEX
L
in
commercial
greenhouses
will
be
limited.
Studies
to
assess
the
effect
of
P.
flocculosa
on
these
organisms
are
not
required.
The
formulants
in
the
end­
products,
SPORODEX
L,
do
not
pose
an
environmental
risk
when
used
at
the
proposed
concentrations
and
application
rate
for
control
of
powdery
mildew
on
roses
and
cucumbers
grown
in
greenhouses.
Consequently,
SPORODEX
L
is
expected
to
pose
little
environmental
risk
when
used
in
accordance
with
the
label
directions.

9.7
Endangered
species
The
Agency
has
no
evidence
to
believe
that
any
endangered
of
threatened
species
will
be
adversely
affected
if
products
containing
Pseudozyma
flocculosa
strain
PF­
A22
UL
are
used
as
labeled
(i.
e.,
greenhouse­
grown
cucumbers
and
roses).
Therefore,
the
Agency
has
determined
that
this
action
will
have
"no
effect"
on
listed
species.
No
specific
labeling
is
required.

9.8
Labels
and
labeling
It
is
Canada/
PMRA's
and
U.
S.
EPA's
position
that
the
labeling
for
products
containing
Pseudozyma
flocculosa
strain
PL­
A22
UL
must
comply
with
the
current
pesticide
labeling
requirements.
SPORODEX
®
L
is
manufactured
following
a
continuous
manufacturing
process
that
does
not
involve
an
intermediate
stand­
alone
technical
product.

9.8.1
End­
use
product
(SPORODEX
®
L)

A
joint
U.
S./
Canada
NAFTA
label
containing
the
necessary
regulatory
language
for
SPORODEX
®
L
is
provided
below.
This
label
was
approved
by
Canada/
PMRA,
June
3,
2002.

DRAFT
U.
S./
CANADA
LABEL
(Front
Panel)

SPORODEX
L
BIOLOGICAL
FUNGICIDE
For
Control
of
Powdery
Mildew
on
Greenhouse
Roses
and
Cucumbers.

COMMERCIAL
READ
THE
LABEL
BEFORE
USING
POTENTIAL
SENSITIZER
ACTIVE
INGREDIENT/
GUARANTEE:
Pseudozyma
flocculosa
strain
PF­
A22
UL
.
.
1.3%
OTHER
INGREDIENTS:
...............................................
98.
7%
TOTAL:
................................................
100.0%
(Contains
a
minimum
of
3
×
10
8
colony
forming
units/
mL)

KEEP
OUT
OF
REACH
OF
CHILDREN
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
45
CAUTION
U.
S.
EPA
Registration
No.:
(Pending
as
File
Symbol
69697­
3)
U.
S.
EPA
Establishment:
69697­
CAN­
001
REGISTRATION
NUMBER
XXXXX
PEST
CONTROL
PRODUCTS
ACT
NetContents:
1
L(
32
U.
S.
floz)

Manufactured
by:
Plant
Products
Co.
Ltd.
314
Orenda
Road
Brampton,
Ontario
L6T
1G1,
Canada
905­
793­
7900
Distributed
by:
Plant
Products
Co.
Ltd.
6160
Riverside
Dr.
Suite
103
Dublin,
OH
43017
Lot
Number:
[XXX]

Date
of
manufacture:
[XXX]
Use
within
3
months
of
the
date
of
manufacture
KEEP
FROZEN
UNTIL
USE
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
46
(Back
Panel)

PRECAUTIONS
/
PRECAUTIONARY
STATEMENTS
HAZARDS
TO
HUMANS
AND
DOMESTIC
ANIMALS.
May
cause
sensitization.
Avoid
contact
with
skin,
eyes
or
clothing.
Avoid
breathing
mist.
Wash
thoroughlywith
soap
and
water
after
handling.

PERSONAL
PROTECTIVE
EQUIPMENT
(PPE):
Applicators
and
other
handlers
must
wear
long­
sleeved
shirt
and
long
pants,
waterproof
gloves
and
shoes
plus
socks.
All
mixers/
loaders
and
applicators
must
wear
a
dust/
mist­
filtering
respirator
(MSHA/
NIOSH
approval
number
prefix
TC21C
or
a
NIOSH
approved
respirator
with
any
N­
95,
R­
95,
P­
95
or
HE
filter
for
biological
products.
Remove
contaminated
clothing
and
follow
manufacturer's
instructions
for
cleaning
/
maintaining
PPE
before
reuse.
If
no
such
instructions
are
available
use
clothing
detergent
and
hot
water
for
cleaning
all
washable
PPE.
Keep
and
wash
PPE
separately
from
other
laundry.

USER
SAFETY
RECOMMENDATIONS:
Users
should
wash
hands
before
eating,
drinking,
chewing
gum,
using
tobacco
or
using
the
toilet.
Remove
PPE
immediately
after
handling
this
product.
Wash
the
outside
of
gloves
before
removing.
As
soon
as
possible,
wash
thoroughly
and
change
into
clean
clothing.

ENVIRONMENTAL
HAZARDS
Do
not
apply
directly
to
water,
or
to
areas
where
surface
water
is
present,
or
to
intertidal
areas
below
the
mean
highwater
mark.
Do
not
contaminate
water
by
cleaning
of
equipment
or
disposal
of
equipment
washwaters.

FIRST
AID
If
on
skin
or
clothing

Take
off
contaminated
clothing.


Rinse
skin
immediately
with
plenty
of
water
for
15
–
20
minutes.


Call
a
poison
control
center
or
doctor
for
treatment
advice.
If
inhaled

Move
person
to
fresh
air.


If
person
is
not
breathing,
call
911
or
an
ambulance,
then
give
artificial
respiration,
preferably
by
mouth­
to­
mouth,
if
possible.


Call
a
poison
control
center
or
doctor
for
treatment
advice.
If
in
eyes

Hold
eye
open
and
rinse
slowly
and
gently
with
water
for
15
­
20
minutes.


Remove
contact
lenses,
if
present,
after
the
first
5
minutes,
then
continue
rinsing
eye.


Call
a
poison
control
center
or
doctor
for
treatment
advice.
If
swallowed

Call
a
poison
control
center
or
doctor
immediately
for
treatment
advice.


Have
person
sip
a
glass
of
water
if
able
to
swallow.


Do
not
induce
vomiting
unless
told
to
do
so
by
the
poison
control
center
or
doctor.


Do
not
give
anything
by
mouth
to
an
unconscious
person.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
47
Seek
medical
attention
IMMEDIATELY
if
irritation
occurs
and
persists
or
is
severe.
Have
container,
label
or
product
name
and
product
registration
number
with
you
when
calling
a
poison
control
center
or
doctor,
or
when
seeking
medical
attention.

TOXICOLOGICAL
INFORMATION
/
NOTE
TO
PHYSICIAN
No
specific
antidote
is
available.
Treat
the
patient
symptomatically.

DIRECTIONS
FOR
USE
In
the
U.
S.
­
It
is
a
violation
of
Federal
law
to
use
this
product
in
a
manner
inconsist
ent
with
its
labeling.
For
any
requirements
specific
to
your
State
or
Tribe,
consult
the
agency
responsible
for
pesticide
regulation.
Do
not
apply
this
product
in
a
way
that
will
contact
workers
or
other
persons,
either
directly
or
thorough
drift.
Only
protected
handlers
may
be
in
the
area
during
application.

In
Canada
­
NOTICE
TO
USER:
This
control
product
is
to
be
use
only
in
accordance
with
the
directions
on
this
label.
It
is
an
offence
under
the
PEST
CONTROL
PRODUCTS
ACT
to
use
a
control
product
under
unsafe
conditions.

In
the
U.
S.
Agricultural
Use
Requirements
Use
this
product
only
in
accordance
with
its
labeling
and
with
the
Worker
Protection
Standard,
40
CFR
Part
170.
This
standard
contains
requirements
for
the
protection
of
agricultural
workers
on
farms,
forests,
nurseries
and
greenhouses,
and
handlers
of
agricultural
pesticides.
It
contains
requirements
for
training,
decontamination,
notification,
and
emergency
assistance.
It
also
contains
specific
instructions
and
exceptions
pertaining
to
the
statements
on
this
label
about
personal
protective
equipment
(PPE),
and
restricted
entry
intervals
(REI).
The
requirements
in
this
box
only
apply
to
uses
of
this
product
that
are
covered
by
the
Worker
Protection
Standard.

Do
not
enter
or
allow
worker
entry
into
treated
areas
during
the
restricted
entry
interval
of
4
hours.

PPE
requirement
for
early
entry
to
treated
areas
that
is
permitted
under
the
Worker
Protection
Standard
and
that
involves
contact
with
anything
that
has
been
treated,
such
as
plants,
soil
or
water,
is
coveralls,
waterproof
gloves
and
shoes
plus
socks.

In
Canada
Do
not
enter
or
allow
worker
entry
into
treated
areas
during
the
restricted
entry
interval
(REI)
of
4
hours.
Workers
can
enter
treated
areas
during
the
REI
if
appropriate
PPE
is
worn,
including
a
long
sleeved
shirt,
long
pants,
shoes
plus
socks
and
waterproof
gloves
as
well
as
a
NIOSH
approved
respirator
with
any
N­
95,
R­
95,
P­
95
or
HE
filter
for
biological
products.

GENERAL
INFORMATION
SPORODEX
L
is
a
naturally
occurring
fungus
which
is
an
antagonist
to
the
powdery
mildew
disease
organism.
SPORODEX
L
is
an
aqueous
liquid
formulation
of
Pseudozyma
flocculosa
formulated
to
control
powdery
mildew
disease
on
the
listed
crops.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
48
APPLICATION
RATES
CROP
DISEASE
RATE
Greenhouse
roses
Greenhouse
cucumbers
Powdery
mildew
disease
In
U.
S.
and
Canada:
500
mL
per
100
L
of
water.
Add
20
mLofanappropriate
wetting
agent
per
100
L
of
spray
mixture.
or
64
U.
S.
fl
oz
per
100
U.
S.
gallons
of
water.
Add
3
U.
S.
fl
oz
of
an
appropriate
wetting
agent
per
100
U.
S.
gallons
of
spray
mixture.

Add
SPORODEX
L
to
water.
Spray
foliage
to
run­
off
at
weekly
intervals,
beginning
when
environmental
conditions
favor
development
of
powdery
mildew,
or
at
first
sign
of
disease.

Apply
up
to
1500
L
of
spray
mixture
per
hectare
(150
U.
S.
gallons
of
spray
mixture
per
acre)
for
cut
roses,
cucumbers
or
about
1000
L
per
hectare
(100
U.
S.
gallons
per
acre)
for
potted
roses.

Maintain
relative
humidity
above
70%
for
12
hours
after
application,
for
example,
by
using
shade
curtain
or
by
applying
SPORODEX
L
late
in
the
day
or
during
prolonged
cloudy
conditions.

NOTE:
Use
of
chemicals
at
the
same
time
as
SPORODEX
L
may
inhibit
this
product's
activity
against
powdery
mildew.
Do
not
tank
mix
SPORODEX
L
with
chemical
pesticides.

SPORODEX
L
has
not
been
tested
for
compatibility
with
all
chemical
and
biological
products
(including
biological
control
insects
and
arthropods)
used
in
greenhouse
production.
For
details
on
compatibility
contact
the
distributor
or
manufacturer,
or
test
effectiveness
on
a
small
number
of
plants
prior
to
commercial
scale
use.

STORAGE
AND
DISPOSAL
Do
not
contaminate
water,
food
or
feed
by
storage
or
disposal.
Pesticide
Storage:
Store
frozen
at
­20

C
(­
4

F)
or
less
and
keep
away
from
food
or
feed.
Keep
product
in
original
container
during
storage
and
keep
container
lid
tightly
closed
when
not
in
use.
This
product
should
be
used
within
3
months
of
the
date
of
manufacture
when
stored
at
­20

C
(­
4

F).
Thaw
at
room
temperature
prior
to
using.

In
the
U.
S.
Pesticide
Disposal:
Wastes
resulting
from
the
use
of
this
product
may
be
disposed
of
on
site
or
at
an
approved
waste
disposal
facility.
Container
Disposal:
Completelyemptypackage
into
application
equipment.
Then
dispose
of
empty
package
in
a
sanitary
landfill
or
by
incineration,
or,
if
allowed
by
State
and
local
authorities,
by
burning.
If
burned,
stay
out
of
smoke.
Do
not
reuse
container.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
49
In
CanadaDISPOSAL
1.
Triple­
or
pressure­
rinse
the
empty
container.
Add
the
rinsings
to
the
spray
to
the
spray
mixture
in
the
tank.
2.
Follow
provincial
instruction
for
any
required
additional
cleaning
of
the
container
prior
to
its
disposal.
3.
Make
the
container
unsuitable
for
further
use.
4.
Dispose
of
the
container
in
accordance
with
provincial
requirements.
5.
For
information
on
the
disposal
of
unused,
unwanted
product,
contact
the
manufacturer
or
the
provincial
regulatory
agency.
Contact
the
manufacturer
and
the
provincial
regulatory
agency
in
case
of
a
spill,
and
for
clean­
up
of
spills.

In
the
U.
S.
­
NOTICE
TO
USER:
Seller
makes
no
warranty,
express
or
implied,
of
merchantability,
fitness
or
otherwise
concerning
the
use
of
this
product
other
than
indicated
on
the
label.
User
assumes
all
risks
of
use,
storage,
or
handling
not
in
strict
accordance
with
label
instructions.

In
Canada
­
NOTICE
TO
BUYER:
Seller's
guarantee
shall
be
limited
to
the
terms
set
out
on
the
label
and,
subject
thereto,
the
buyer
assumes
the
risk
to
persons
or
property
arising
from
the
use
or
handling
of
this
product
and
accepts
the
product
on
that
condition.

SPORODEX
L
is
a
trademark
of
Plant
Products
Co.
Ltd.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
50
9.8.2
Manufacturing­
use
product
Manufacturing
Use
Label
NOT
TO
BE
USED
DIRECTLY
FOR
TREATMENT
OF
PESTS
PSEUDOZYMA
FLOCCULOSA
Strain
PF­
A22
Technical
Grade
Active
Ingredient
for
Manufacturing
Use
Only.

ACTIVE
INGREDIENT:
Pseudozyma
flocculosa
StrainPF­
A22UL
...............
9.
00%
OTHER
INGREDIENTS:
..............................................
91.
00%
TOTAL:
................................................
100.0%
(Contains
a
minimum
of
2
×
10
9
colony
forming
units/
mL)

KEEP
OUT
OF
REACH
OF
CHILDREN
CAUTION
U.
S.
EPA
Registration
No.:
(Pending
as
File
Symbol
69697­
R)
U.
S.
EPA
Establishment:
69697­
CAN­
001
Net
Contents:
(XX)

Manufactured
by:
Plant
Products
Co.
Ltd.
314
Orenda
Road
Brampton,
Ontario
L6T
1G1,
Canada
905­
793­
7000
Lot
Number:
[XXX]

Date
of
manufacture:
[XXX]
Use
within
3
months
of
the
date
of
manufacture
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
51
PRECAUTIONARY
STATEMENTS
HAZARDSTOHUMANS
ANDDOMESTICANIMALS.
CAUTION.
Maycause
sensitization.
Avoid
contact
with
skin,
eyes
or
clothing.
Avoid
breathing
mist.
Wash
thoroughly
with
soap
and
water
after
handling.

PERSONAL
PROTECTIVE
EQUIPMENT
(PPE):
Wear
a
dust/
mist­
filtering
respirator
(MSHA/
NIOSH
approval
number
prefix
TC­
21C),
or
a
NIOSH
approved
respirator
with
any
N­
95,
R­
95,
P­
95
or
HE
filter
for
biological
products.
Remove
contaminated
clothing
and
follow
manufacturer's
instructions
for
cleaning
/
maintaining
PPE
before
reuse.
If
no
such
instructions
are
available
use
clothing
detergent
and
hot
water
for
cleaning
all
washable
PPE.
Keep
and
wash
PPE
separately
from
other
laundry.

ENVIRONMENTAL
HAZARDS
Do
not
discharge
effluent
containing
the
product
into
lakes,
streams,
ponds,
estuaries,
oceans,
or
other
waters
unless
in
accordance
with
the
requirements
of
the
National
Pollutant
Discharge
Elimination
System
(NDPES)
permit
and
the
permitting
authority
has
been
notified
in
writing
prior
to
discharge.
Do
not
discharge
effluent
containing
this
product
to
sewer
systems
without
previously
notifying
the
local
sewage
treatment
plant
authority.
For
guidance,
contact
your
State
Water
Board
or
Regional
Office
of
the
EPA.

FIRST
AID
If
on
skin
or
clothing

Take
off
contaminated
clothing.


Rinse
skin
immediately
with
plenty
of
water
for
15
–
20
minutes.


Call
a
poison
control
center
or
doctor
for
treatment
advice.
If
inhaled

Move
person
to
fresh
air.


If
person
is
not
breathing,
call
911
or
an
ambulance,
then
give
artificial
respiration,
preferably
by
mouth­
to­
mouth,
if
possible.


Call
a
poison
control
center
or
doctor
for
treatment
advice.
If
in
eyes

Hold
eye
open
and
rinse
slowly
and
gently
with
water
for
15
­
20
minutes.


Remove
contact
lenses,
if
present,
after
the
first
5
minutes,
then
continue
rinsing
eye.


Call
a
poison
control
center
or
doctor
for
treatment
advice.
If
swallowed

Call
a
poison
control
center
or
doctor
immediately
for
treatment
advice.


Have
person
sip
a
glass
of
water
if
able
to
swallow.


Do
not
induce
vomiting
unless
told
to
do
so
by
the
poison
control
center
or
doctor.


Do
not
give
anything
by
mouth
to
an
unconscious
person.

Have
container,
label
or
product
name
and
product
registration
number
with
you
when
calling
a
poison
control
center
or
doctor,
or
when
seeking
medical
attention.

DIRECTIONS
FOR
USE
It
is
a
violation
of
Federal
law
to
use
this
product
in
a
manner
inconsistent
with
its
labeling.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
52
FOR
MANUFACTURING
USE
ONLY.
Only
for
formulation
into
end­
use
products,
for
use
to
control
plant
diseases
in/
on
agricultural
commodities.
Not
for
direct
treatment
of
pests.
Do
not
use
from
damaged,
punctured
or
unsealed
containers
STORAGE
AND
DISPOSAL
Do
not
contaminate
water,
food
or
feed
by
storage
or
disposal.
Pesticide
Storage:
Store
refrigerated
and
keep
away
from
food
or
feed.
Pesticide
Disposal:
Wastes
resulting
from
the
use
of
this
product
may
be
disposed
of
on
site
or
at
an
approved
waste
disposal
facility.
Container
Disposal:
Triple
rinse
(or
equivalent).
Then
offer
for
recycling
or
reconditioning,
or
puncture
and
dispose
of
in
a
sanitary
landfill,
or
incineration,
or,
if
allowed
by
state
and
local
authorities,
by
burning.
If
burned,
stay
out
of
smoke.
Completely
empty
package
into
application
equipment.

NOTICE
TO
USER:
Seller
makes
no
warranty,
express
or
implied,
of
merchantability,
fitness
or
otherwise
concerning
the
use
of
this
product
other
than
indicated
on
the
label.
User
assumes
all
risks
of
use,
storage,
or
handling
not
in
strict
accordance
with
label
instructions.
Product
Monograph
­
Pseudozyma
flocculosa
strain
PF­
A22
UL
53
Chapter
10
List
of
abbreviations
AUDPC
area
under
disease
pressure
curve
bw
body
weight
CFU
colony
forming
units
DNA
deoxyribonucleic
acid
dw
dry
weight
EPA
Environmental
Protection
Agency
(U.
S.)
FFDCA
Federal
Food
Drug
and
Cosmetic
Act
(U.
S.)
FIFRA
Federal
Insecticide
Fungicide
Rodenticide
Act
(U.
S.)
FQPA
Food
Quality
Protection
Act
(U.
S.)
EP
end­
use
product
IPM
integrated
pest
management
KTS
killed
test­
substance
LD50
lethal
dose
50%
MA
Martin's
agar
MAS
maximum
average
score
MIS
maximum
irritation
score
MPCA
microbial
pest
control
agent
MRL
maximum
residue
limit
(Canada)
NC
naive
control
PCA
plate
count
agar
PCR
polymerase
chain
reaction
PDA
potato
dextrose
agar
PMRA
Pest
Management
Regulatory
Agency
(Canada)
PPE
personal
protective
equipment
RAMS
random
amplified
microsatellites
REI
restricted
entry
interval
RH
relative
humidity
TGAI
technical
grade
of
the
active
ingredient
TS
test
substance
U.
S.
United
States
of
America
U.
S.
EPA
United
States
Environmental
Protection
Agency
YM
yeast
malt
agar