Regulation (EU) No 528/2012
concerning the making available on the
market and use of biocidal products
Evaluation of active substances
Assessment Report
Pythium oligandrum Strain M1
Product-type 10
(Masonry preservative)
January 2015
The Czech Republic
Pythium oligandrum M1
Product-type 10
January 2015
CONTENTS
1. STATEMENT OF SUBJECT MATTER AND PURPOSE ................................. 3
1.1. Procedure followed ............................................................................................................................ 3
1.2. Purpose of the assessment report ............................................................................................. 3
2. OVERALL SUMMARY AND CONCLUSIONS............................................................... 4
2.1. Presentation of the Active Substance ....................................................................................... 4
2.1.1. Identity, Physico-Chemical Properties & Methods of Analysis ............................................. 4
2.1.2. Intended Uses and Efficacy ................................................................................................................ 4
2.1.3. Classification and Labelling ................................................................................................................ 5
2.2. Summary of the Risk Assessment .............................................................................................. 5
2.2.1. Human Health Risk Assessment ....................................................................................................... 5
2.2.1.1. Hazard identification ..................................................................................................................... 5
2.2.1.2. Effects assessment ........................................................................................................................ 5
2.2.1.3. Exposure assessment ................................................................................................................... 6
2.2.1.4. Risk characterisation ..................................................................................................................... 7
2.2.2. Environmental Risk Assessment ....................................................................................................... 9
2.2.2.1. Fate and distribution in the environment ............................................................................. 9
2.2.2.2. Effects assessment ...................................................................................................................... 10
2.2.2.3. PBT and POP assessment .......................................................................................................... 11
2.2.2.4. Exposure assessment ................................................................................................................. 12
2.2.2.5. Risk characterisation ................................................................................................................... 12
2.2.3. Assessment of endocrine disruptor properties ......................................................................... 12
2.3. Overall conclusions .......................................................................................................................... 13
2.4. List of endpoints ................................................................................................................................ 13
APPENDIX I: LIST OF ENDPOINTS .............................................................. 14
APPENDIX II: LIST OF INTENDED USES .................................................... 24
APPENDIX III: LIST OF STUDIES ................................................................. 25
2
Pythium Oligandrum M1
Product-type 10
January 2015
1. STATEMENT OF SUBJECT MATTER AND PURPOSE
1.1. Procedure followed
This assessment report has been established as a result of the evaluation of the active
substance Pythium oligandrum Strain M1 as product-type 10, carried out in the context of
the work programme for the review of existing active substances provided for in Article 89 of
Regulation (EU) No 528/2012, with a view to the possible approval of this substance.
On 12 July 2005, the CZ competent authorities received a dossier from the applicant. The
Rapporteur Member State accepted the dossier as complete for the purpose of the
evaluation on 4 August 2010.
On 8 November 2011 the Rapporteur Member State submitted to the Commission and the
applicant a copy of the evaluation report, hereafter referred to as the competent authority
report.
In order to review the competent authority report and the comments received on it,
consultations of technical experts from all Member States (peer review) were organised by
the Agency. Revisions agreed upon were presented at the Biocidal Products Committee and
its Working Groups meetings and the competent authority report was amended accordingly.
1.2. Purpose of the assessment report
The aim of the assessment report is to support the opinion of the Biocidal Products
Committee and a decision on the approval of Pythium oligandrum M1 for product-type 10,
and, should it be approved, to facilitate the authorisation of individual biocidal products. In
the evaluation of applications for product-authorisation, the provisions of Regulation (EU) No
528/2012 shall be applied, in particular the provisions of Chapter IV, as well as the common
principles laid down in Annex VI.
For the implementation of the common principles of Annex VI, the content and conclusions
of this assessment report, which is available from the Agency web-site shall be taken into
account.
However, where conclusions of this assessment report are based on data protected under
the provisions of Regulation (EU) No 528/2012, such conclusions may not be used to the
benefit of another applicant, unless access to these data for that purpose has been granted
to that applicant.
3
Pythium Oligandrum M1
Product-type 10
January 2015
2. OVERALL SUMMARY AND CONCLUSIONS
2.1. Presentation of the Active Substance
2.1.1. Identity, Biological and Physico-Chemical Properties and Methods of Analysis
Active substance is concentrate of spores of Pythium oligandrumStrain M1 (Technical grade
Pythium oligandrum) consisting of the remnants of the growing medium (about 95%),
mycelium and spores of Pythium oligandrum (2.5 x 106 – 10 x 106 oospores/g per a.s.).
Pythium oligandrum belongs to the Oomycota class classified as a part of newly created
kingdom Stramenopila (syn. Chromista). Pythium oligandrum is a living organism, its dormant
spores can survive temperatures under 60°C, but do not grow at temperatures > 37°C in
tissues of animal origin. Pythium oligandrum is naturally occurring in soil where it colonizes
the rhizosphere of plants. It requires a humid environment and has a minimum growth
temperature of 7ºC, optimum around 30 ºC.
The M1 strain of Pythium oligandrum was deposited in American Type Culture Collection
(ATCC) under reference number ATCC 38472 and in Czech Collection of micro-organisms under
mark M1.
Technical grade Pythium oligandrum (further on tgPO) is fine whitish powder, dispersible in
water. Quality management of the manufacturing process ensures that no toxins or pathogens
relevant for human health are present in the technical grade active ingredient.
Analysis
The micro-organism could be identified by light microscopy – it has well distinguishable spinywalled oospores- and can also be identified using PCR. Oospores are counted in Buerker's
chamber. PCR enables differentiation between different Pythium oligandrum Strain M1 strains.
Coupling PCR with other methods provides a battery of methods enabling identification of
Pythium oligandrum Strain M1 at strain level. The methods are listed and described in DOC IIA
and their detailed descriptions are given in the corresponding DOC III section or annex I to
DOC IIA. Strain level characterisation can be done by a combination of PCR, HPLC MS/MS and
methods based on biological characteristics (e.g. multiplication rate under specific conditions).
2.1.2. Intended Uses and Efficacy
Pythium oligandrum is a mycoparasite. With its hyphae, Pythium oligandrum penetrates cells
of the target organisms (mould or yeast), drawing from it necessary substances for its
nourishment. As soon as the nourishment source is exhausted, the microorganism will
transform into a spore stage and wait for more favourable conditions.
The assessment of the biocidal activity of the active substance demonstrates that it has a
sufficient level of efficacy against moulds on masonry and the evaluation of the summary data
provided in support of the efficacy of the accompanying product, establishes that the product
may be expected to be efficacious.
Efficacy tests were conducted with the product Biorepel: walls heavily infected by saprophytic
molds, monoculture of Aspergillus terreus or mixture of A. niger, A. terreus and A.
auranogriseum were sucessfuly cleansed in 4 weeks by a single treatment with suspension
containing 200mg TGAI/L water, one litre of suspension per 5 m2. Moulds on wooden carriers
were not eliminated even after 6 weeks.
The field of use envisaged is indoor application against undesirable moulds on walls and
plasters, both as curative and preventative treatment by brushing/rolling.
In addition, in order to facilitate the work of Member States in granting or reviewing
authorisations, the intended uses of the substance, as identified during the evaluation process,
are listed in Appendix II.
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Pythium Oligandrum M1
Product-type 10
January 2015
2.1.3. Classification and Labelling
The substance is a microorganism and hence is covered neither by Directive 67/548/EEC nor
by the CLP regulation. It produces no metabolite classified as hazardous according to Directive
67/548/EE or the CLP regulation.
The classification and labelling for Pythium oligandrum Strain M1 according to Regulation (EC)
No 1272/2008 (CLP Regulation) is not required as the active substance is a microorganism not
covered by this regulation. The classification according to Directive 67/548/EE or the CLP
regulation could only concern chemical substances produced by the Pythium oligandrum Strain
M1 or already contained in it. No substance that could be classified as hazardous was identified
and the toxicological studies provided no indication of toxicity.
Considering that all microbials should be regarded as potential sensitisers, the agreed warning
phrase on the product label is: “Microorganisms may have the potential to provoke sensitising
reactions”.
2.2. Summary of the Risk Assessment
2.2.1. Human Health Risk Assessment
It is noted that EFSA in their expert opinion on Pythium oligandrum Strain M1 published in
2013 concluded that a derivation of reference values is not necessary and no quantitative risk
assessment is required. In the light of this and the conclusions from the data on which this
report is based measures protecting humans, animals and the environment following form the
hazard identification has been preferred in this report. The human exposure calculations and
their comparison to reference values are provided in this report for information only.
Toxicokinetics and metabolism
Pythium oligandrum produces several substances that play a role in the organism’s mode of
action. None of these substances are considered to be of toxicological concern.
Dermal absorption of particulate material is not expected.
2.2.1.1. Hazard identification
No systemic effects have been observed in any of the toxicological studies provided by the
applicant. Two studies on skin irritation and standard test of acute eye irritation provide
evidence of slight irritating properties of the TGAI.
2.2.1.2. Effects assessment
Systemic effects
As no systemic effects have been observed after neither single nor repeated dosing, the top
doses of the technical active substance are indicated as toxicity level 0 (TL0, NOAEL) values
(Table 2.1). Due to the lack of toxicity derivation of reference values and hence, a quantitative
risk assessment is not required. It is noted that the same conclusion has been reached by the
pesticide peer review on Pythium oligandrum Strain M1 M1 published in 20131. Pythium
oligandrum does not produce any known metabolites considered to be of toxicological concern:
genotoxicity, carcinogenicity and reproductive toxicity testing of specific metabolites therefore
has not been conducted.
1
EFSA Journal 2013;11(1):3034
5
Pythium Oligandrum M1
Table 2.1
Route
animal
oral gavage
rat, mouse
oral gavage
mouse
dermal
rat
inhalation
rat
4 hour
exposure
oral, in diet
rat, 70 days
Product-type 10
TL0 values
Material administered
active substance
in 12.5% suspension
active substance,
sterilised, suspended
active substance
moistened (1.5ml/g)
24 hour
semiocclusion
active substance
5 mg/L air
GM 7.45 µm
active substance
0.8, 2.4, 4 % in diet
January 2015
TL0 values
mg a.s./kg
bw
single dose
>5000
single dose
>20 000
single dose
>5000
Observation
period
Examination
14 days
clinical,
necropsy
mortality
behaviour
weight, clinical,
local signs
necropsy
single dose
>600
14 days
repeated
daily dose **
>4000
70 days
10 hours
14 days
weight, clinical signs,
behaviour,
necropsy,
histopathology of
respiratory system.
weight, clinical,
behavioural **
haematology,
necropsy, organ
weights,
histopathology
* active suspension has been filtered, resulting count of propagules in the ip dose = 1.8 x 10 5
/kg bw, corresponding to active substance dose of 360 mg/kg bw.
** for an average daily consumption of diet 100 g/kg bw; increased body weight in exposed
animals and differences in some behavioural tests are not indicative of toxicity.
Infectivity and pathogenicity
The active substance is not considered as infective or pathogenic. Maximum tolerated
temperature of 37 ºC, determined in the in-vitro study on growth of Pythium oligandrum in
tissues of animal origins, prevents deep infections in tissues of mammals.
Local effects
Two studies on the skin irritation and standard test of acute eye irritation provide evidence of
slight irritating properties of the neat a.s.
No immediate or delayed reactions were observed after 24 hour exposure of 1/10 of skin
surface (cca 10 cm2) to concentrated suspension of 5000 mg of the active substance. Local
concentration of the active substance is estimated to have been at least 500 mg/cm 2.
Human data: No clinical cases or sensitisation / allergic reactions were noted in the facilities of
the applicant.
2.2.1.3. Exposure assessment
The human exposure calculations and their comparison to reference values are provided only
for information. Exposure to a concentrated product occurs during industrial manufacture of
biocidal product, which takes place once a year, and during mixing of the active suspension by
professional operators or amateur users. Exposure is possible only in the phase of manual
pouring of the active substance or of mixed product (max. 20% a.s.). Inhalation and dermal
deposition of dust are the relevant routes of exposure.
6
Pythium Oligandrum M1
Product-type 10
January 2015
Exposure to an active suspension occurs during application of the suspension (100 – 200 mg
a.s./L) by manual tools (brushing). Inhalation and dermal deposition of the liquid aerosol are
the relevant routes of exposure.
The exposure calculations and the resultant values are provided in section 3.2 of DOC IIB.
The exposure values are provided only for information.
2.2.1.4. Risk characterisation
Due to the lack of toxicity, derivation of reference values and human exposure calculation are
considered unnecessary. Consequently, a quantitative risk assessment is not considered
necessary and is included only for information in the CAR. The conclusions on measures to
protect man, animals and the environment follow from consideration of the relevant routes of
exposure and the hazard identification.
Critical endpoints and routes of exposure
The toxicologically relevant endpoints for the active substance are summarised in
Document IIA, Chapter 3.
Dermal absorption is highly unlikely due to the nature of the active substance. Maximum
tolerated temperature of 37 ºC prevents deep infections in tissues of mammals.
No systemic toxic effects or signs of infectivity were observed in the toxicity tests.
Sensitisation has not been observed in exposed workers but in the absence of a reliable test on
sensitisation, as for other microorganisms, it has not been ruled out.
Relevant exposure routes are inhalation and dermal exposure to the powder of active
substance or product, and to liquid aerosol generated e.g. in the brushing or mixing/loading
phase.
Industrial formulation of the active substance
This operation is conducted once per year by professionals: exposure is considered to be
accidental. The estimated doses are compared with the acute NOAELs for information only.
Inhalation and dermal exposure to powder during manual loading of mixing and packaging
machines is estimated to result in the total daily (external) dose of 6.52 mg a.s./kg bw, MOE
= 5000/6.52 = 767. The estimate of dermal deposition is 0.44 mg of a.s./cm2 , < 1/1000 of
no-effect local concentration of 500 mg/cm2.
With respect to possible sensitising potency of the active substance, both inhalation and
dermal exposures of workers not using PPE may be too high during manual pouring of the
active substance and of the product. Thus, when performing this task appropriate PPE
including gloves and RPE should be worn.
Application of the biocidal product
Total daily exposure is estimated in the first tier calculation to be ≤ 0.084 mg a.s./kg bw for
professional operators. Using subacute TL0 (oral) of 4000 mg/kg bw as a measure, MOE =
4000/0.084 > 4x 104. Margins over exposure are even higher for non-professional users (>105
) for the first tier calculation.
Since Pythium oligandrum Strain M1 is a microorganism the risk of sensitizing effects during
mixing and loading and its application is assessed.
The duration of 150 minutes proposed in User guidance p.47 for professional brush-painting of
wooden objects provides a conservative estimate of exposure duration of the proposed use.
The non-professional exposure is assumed to be of shorter duration, usually not exceeding 60
minutes.
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Pythium Oligandrum M1
Product-type 10
January 2015
For non-professionals, the risk of exposure to the undiluted product at the loading/mixing
stage can be mitigated by using water soluble packaging. This type of packaging is considered
as a necessary risk mitigation measure for the non-professionals, unless reliable data or
scientifically sound justification showing that the exposure to the undiluted product poses no
unacceptable risk are provided at the product authorisation stage.
For the professional users an alternative to water soluble packaging is the use of appropriate
personal protective equipment (PPE) including gloves, long sleeved coveralls and respiratory
protective equipment (RPE, dust mask).
Dermal exposure during the application phase is considered as posing no unacceptable risk
either to professionals or non-professionals due to the high dilution rate of the working solution
This conclusion is supported by the fact that the representative product contains only 20% of
technical grade active ingredient (TGAI). TGAI contains only 5% of oo-spores and mycelium
(the rest are remnants of nutrients and agar from its manufacture). Thus, in total the biocidal
product contains only 1% of “pure” Pythium oligandrum. In reality the exposure of both
professionals and non-professionals takes place only during the brushing phase when the
product is diluted with water provided that the exposure during the mixing phase is prevented
water soluble packaging. The worst case application solution then contains 1g of Biorepel
product in 1 liter of water. The concentration of the “pure” a.i. in the application solution is
therefore 0.001% which is considered sufficiently low. This, combined with the fact that
Pythium oligandrum Strain M1 does not grow in animal tissue at 37oC and cannot therefore
elicit an allergic reaction by continually producing chemical substances during its growth, adds
weight to the conclusion that the brushing application does not pose unacceptable risk to
either non-professionals or professional users.
The by-stander inhalation exposure to Pythium oligandrum Strain M1 from the plant protection
product (PPP), Polyversum application was compared to the exposures of users and bystanders
during biocidal product application by brushing. According to the PPP assessment report (RMS
SE), by-standers were considered to be exposed for 30 minutes to spray drift when walking
next to a field being treated, considered to be 10 m from the sprayer. According to the
assessment, the estimated inhaled dose was 0.000021 mg/kg bw of the active. The exposure
of professional and non-professional users, as well as by-standers during the brushing
application is considered to be even lower, as the in-use concentration is more than fourfold
lower, and exposure from a spray drift is a worst case compared to brushing. Consequently,
the exposure via inhalation to Pythium oligandrum Strain M1 during the brushing application is
considered as safe both for professionals and non-professionals as well as for by-standers.
In summary, considering the hazard profile and the exposure, even without the use of PPE the
brushing application does not pose unacceptable risk to either by-standers, non-professional or
professional users.
Indirect exposure
Indirect exposure can accidentally take place by skin contact with freshly treated wall and/or
inhalation of the particles of active substance released from the wall after drying. The estimate
of exposure due to contact with the freshly treated wall is lower than that due to hand-brush
application by non-professional users.
Regarding the inhalation of the active particles released from the wall after it has dried the
factors influencing its release to the air have been considered. It is assumed that the exposure
via inhalation is possible only if the active substance is released form the wall surface due to a
physical disturbance and is kept in the air by a sufficiently intense air movement. These factors
are regarded as negligible for the Pythium Oligandrum Strain M1 intended use – indoor leading to negligible exposure compared to the inhalatory exposure to Pythium oligandrum
naturally present in the soil dust.
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Pythium Oligandrum M1
Product-type 10
January 2015
Conclusion
The quantitative risk assessment was provided only for information as the toxicological studies
identified no hazard to human health. As a precautionary measure applied to microorganisms
in general, it has to be considered that Pythium oligandrum Strain M1 may cause a
sensitization reaction. Therefore, significant exposure to the technical grade active ingredient
or the undiluted product by industrial workers and/or professionals should be prevented by the
use of appropriate PPE including RPE, or other RMM. Exposure of professionals and nonprofessionals to the aqueous suspension of the biocidal product during application is
considered to be acceptable without the use of PPE.
Indirect exposure due to the proposed use is considered to be acceptable. The combined
exposure is considered to be acceptable for both professionals and non-professionals.
This risk assessment for the proposed use of the technical grade Pythium oligandrum Strain
M1 in the Product Type 10 has demonstrated that there are no concerns regarding human
health.
2.2.2. Environmental Risk Assessment
2.2.2.1. Fate and distribution in the environment
Pythium oligandrum Strain M1 is intended for preventive and /or curative treatment of walls
against molds indoors only. Mostly, the applied Pythium oligandrum Strain M1 is left on the
wall even after the mould has been destroyed. Should the conditions under which humidity and
hence moulds re-occur the a.s. will “wake up” and feed on the moulds as described in the
section 1.3. For this reason the place affected shall not be rinsed after the application, hence
the emissions of Pythium oligandrum Strain M1 to the environment will be negligible. The
emissions to the STP are semi-quantified in DOC IIB with the conclusion that the STP exposure
to Pythium oligandrum Strain M1 due to application of the biocidal product Biorepel is
negligible compared to its exposure to Pythium oligandrum Strain M1 washed to STP from soil
during rain.
Biotic degradation
Pythium oligandrum Strain M1 is a micro-organism naturally occurring in the environment,
mainly in soil. It is not biodegradable in the meaning of degradation known in organic chemical
substances.
Pythium oligandrum Strain M1 occurs in soil where it colonizes the rhizosphere of plants. Its
concentration in soil depends on a number of factors primarily including pH, temperature and
occurrence of fungi it can feed on. The optimal conditions for Pythium oligandrum Strain M1
abundance in soil are provided in section 4.1. of DOC IIA.
Abiotic degradation
Hydrolysis: Pythium oligandrum Strain M1 is a micro-organism, hydrolysis is not possible.
Photolysis: Pythium oligandrum Strain M1 is a micro-organism, photolysis is not possible.
Phototransformation in air: In case of Pythium oligandum Strain M1, air is a transport
medium for oospores, a growth itself does not occur in air.
Distribution
Pythium oligandrum Strain M1 is a naturally occurring soil micro-organism. The proposed
indoor application of products based on Pythium oligandrum Strain M1 can have significant
effects on its fate, behavior and distribution in the environment. Pythium oligandrum Strain M1
occurs in relatively high densities in many agricultural soils “feeding on” many soil fungi.
Pythium oligandrum occurs mostly in rhizosphaere of soils of disturbed sites (agricultural land).
9
Pythium Oligandrum M1
Product-type 10
January 2015
It was observed that Pythium oligandrum population decreased with time after being
transplanted in sterile soil. Takenaka et all (2008). Madsen (2004) reported concentration
between 4 and 26 oospores/g soil in Danish soils, Al-Rawahi and Hancock (1997) reported 89151 oospores of Pythium oligandrum/g of raw field soil in Pakistan. Pythium oligandrum Strain
M1 does not occur in aquatic environment which is considered as unfavorable for its growth
(Foley and Deacon (1985), Klaban (2013).
Adsorption onto/desorption from soil
No adsorption or desorption, as known in organic chemical substances, occur in Pythium
oligandum Strain M1.
Accumulation
Pythium oligandrum Strain M1 is originally a soil micro-organism and it does not accumulate
nor multiplicates in living organisms. With its hyphae, the fungal organism Pythium oligandrum
penetrates cells of the parasite (mould or yeast), drawing from it necessary substances for its
nourishment. As soon as the nourishment source is exhausted, the fungus will change into the
spore stage. It does not reproduce in haematothermal organisms; temperatures above 37 °C
do not suit it (Kratochvílová, 2006).
2.2.2.2. Effects assessment
Acute and chronic toxicity to fish
No effects on fish were observed in the acute toxicity fish study. Therefore the LC50 value
could not be calculated. No fish deaths or morphological changes were observed at
concentration 100 mg active substance per litre (top dose). Therefore, the acute NOEC could
be considered as > 100 mg/L.
Acute and chronic toxicity to invertebrates
No adverse effects on invertebrates were identified by a literature research. Since direct
release of the active substance to the aquatic environment is not expected to occur, a test on
invertebrates was waived.
Growth inhibition on algae
Pythium oligandrum Strain M1 is ubiquitous as a naturally-occurring soil colonizer whose
modes of action that is not consistent with toxicity or pathogenicity to algae. This is confirmed
by an extensive literature search where no adverse effects on algae were identified. For more
details and references supporting these arguments see section 4.1 of Document IIA. Since
direct release of the active substance to the aquatic environment is not expected to occur, a
test on algae was waived.
Inhibition of activated sludge respiration rate
Pythium oligandrum Strain M1 is ubiquitous as a naturally-occurring soil colonizer whose level
in the STP will not significantly increase with the use of products that contain this strain. For
information, the added STP concentration is quantified in DOC IIB using an emission scenario
for indoor painting. In the calculation, parameters selected by expert judgement were used as
no harmonized guidance is available. The thus obtained number of oospores is significantly
lower than the number of oospores assumed to enter the STP with soil due to commonly
performed anthropogenic activities. It is further noted that Pythium oligandrum Strain M1 has
modes of action that are not consistent with toxicity or pathogenicity to bacteria occurring in
the STP sludge. This is confirmed by an extensive literature search where no adverse effect on
aquatic microorganisms was identified. Furthermore, water is not the environment, where
Pythium oligandrum could multiply (for more details and references supporting these
arguments see section 4.1 in DOC IIA). Thus, no unacceptable risk to STP is envisaged due to
the proposed use of Pythium oligandrum Strain M1.
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Pythium Oligandrum M1
Product-type 10
January 2015
Atmosphere
Air might be a transport medium for oospores but probably not an environment for growth of
the Pythium oligandrum Strain M1.
Terrestrial compartment
Pythium oligandrum Strain M1 naturally occurs in various soil types in which it may persist.
The conditions for Pythium oligandrum Strain M1 occurrence in the terrestrial compartment are
provided in detail in section 4.1 of DOC IIA. Its mobility and persistence in soil is primarily
dependent on the presence of fungus which, in turn, are dependent on the presence of plants.
The conditions for Pythium oligandrum Strain M1 occurrence in the terrestrial compartment are
provided in greater detail in DOC IIA.
It has no dangerous effects on bees and other arthropods, and soil organisms (earthworms are
in contact with the active substance, which is naturally occurring in soil, for their whole
lifecycle without any adverse effects).
The risk for plants is provided in the assessment report elaborated in support of Pythium
oligandrum Strain M1 inclusion to Annex I to Directive 91/414 EC. The active substance is not
harmful to plants.
Determination of predicted no effect concentrations for aquatic compartment
Due to the substance nature, no data for PNEC calculation are available. If used as specified,
the active substance shall not enter the aquatic environment.
Determination of predicted no effect concentrations for terrestrial compartment
Due to the substance nature, no data for PNEC calculation are available.
Determination of predicted no effect concentrations for STP micro-organism
Due to the substance nature, no data for PNEC calculation are available. If used as specified,
the active substance shall not enter sewage treatment plants.
2.2.2.3. PBT and POP assessment
P assessment
Pythium oligandrum Strain M1 is a microorganism and therefore a PBT assessment, as it is
normally performed for chemical substances, is irrelevant. However, according to Addendum to
TNsG on Data Requirements for microorganisms appropriate information on the persistence of
the microorganism in all the relevant compartments has to be given, unless it can be justified
that exposure of the particular environmental compartment to the micro-organism is unlikely
to occur. Since Pythium oligandrum Strain M1 is intended for indoor use only, direct emissions
to any environmental compartment are unlikely to occur. The exposure of STP is semiquantified in DOCIIB with the conclusion that it is negligible. It follows that indirect exposure of
surface water or soil is also negligible. For more information on possible Pythium oligandrum
Strain M1 survival or multiplication in these environmental compartments (see section 4.1. of
DOC IIA). In the light of this, it can be concluded that persistence or multiplication of Pythium
oligandrum Strain M1 in the environment due to the proposed use is of no concern and no
further studies are needed on this endpoint.
B assessment
Pythium oligandum Strain M1 is a micro-organism, it does not meet criteria for being classified
as bioaccumulative. The strain M1 has been shown not to be able to grow at 37 ºC (see
B.6.2.2) (
), thus preventing multiplication in mammals.
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Pythium Oligandrum M1
Product-type 10
January 2015
T assessment
Pythium oligandum Strain M1 is a micro-organism and thus cannot be toxic per se. Regarding
chemical substances Pythium oligandrum Strain M1 contains or produces, their toxicity was
tested in toxicological studies (see section 3.10. of DOC IIA). These studies show that Pythium
oligandrum Strain M1 contains no substances resulting in it being classified as toxic and T
classification is therefore excluded.
Overall assessment
Pythium oligandum Strain M1 is neither a PBT/vPvB nor is it POP.
2.2.2.4. Exposure assessment
Aqueous medium
The proposed use as a fungicide intended for the treatment of walls indoors ensures that
aquatic environment exposure is considered to be negligible. For more information including
semi-quantitative assessment of Pythium oligandrum Strain M1 emissions to STP on its
application see section 3.3. in DOC IIB.
Terrestrial medium
The proposed use as a fungicide intended for the treatment of walls indoors ensures that there
are no direct emissions to soil, and neither direct or indirect emissions to STPs. Theoretically,
Pythium oligandrum Strain M1 could enter the terrestrial compartment when the STP sludge is
applied on soil. However, only negligible amount of Pythium oligandrum Strain M1 can get to
the STP following Biorepel application (for semi-quantitative assessment of Pythium
oligandrum Strain M1 emissions to STP on its application see section 3.3. in DOC IIB).
Furthermore STP sludge is treated befeore it is applied onto the soil thus preventing any viable
Pythium oligandrum Strain M1 oospores from getting to the soil (see section 4.1 in DOC IIA for
details).
Air
There is no direct emission to the atmosphere due to Pythium oligandrum Strain M1
application.
Exposure scenarios
Pythium oligandum is part of the environment. If used as specified there are no significant
emissions to the environment. However semi quantitative assessment of Pythium oligandrum
Strain M1 emissions to STP is provided in section 3.3. in DOC II B with the conclusion that the
emissions to STP due to Biorepel application are negligible and significantly lower than the
emissions due to rain.
2.2.2.5. Risk characterisation
Pythium oligandum Strain M1 is a natural part of the environment; no dangerous properties for
the environment were identified during the risk assessment. Thus, if used as specified in this
Pythium oligandrum Strain M1 poses no unacceptable risk to the environment.
2.2.3. Assessment of endocrine disruptor properties
Pythium oligandrum Strain M1 or substances it produces are not endocrine disruptors. No
toxicity due to mechanism disrupting the endocrine signalling or glands was observed in the
submitted toxicological studies nor any evidence exists suggesting a potential for endocrine
disruption by Pythium oligandrum Strain M1 in humans following long term manufacture and
usage.
12
Pythium Oligandrum M1
Product-type 10
January 2015
2.3. Overall conclusions
The outcome of the assessment for Pythium oligandrum Strain M1 in product-type 10 is
specified in the BPC opinion following discussions at the BPC 8 meeting of the Biocidal Products
Committee (BPC). The BPC opinion is available from the ECHA web-site.
2.4. List of endpoints
The most important endpoints, as identified during the evaluation process, are listed in
Appendix I.
13
Pythium Oligandrum M1
Product-type 10
January 2015
Appendix I: List of endpoints
Chapter 1:
Identity, Biological Properties, Details of Uses, Further Information, and Proposed
Classification and Labelling
Active micro-organism
Function (e.g. fungicide)
Pythium oligandrum, M1 strain, Technical grade
fungicide
Rapporteur Member State
Czech Republic
Identity (Annex IIA, point II.)
Name of the organism:
Taxonomy:
Species, subspecies, strain:
identification:
Culture collection:.
Minimum and maximum concentration of the
micro-organism used for manufacturing of the
formulated product (cfu/g; cfu/L, etc.):
Identity and content of relevant impurities in
thetechnical grade micro-organism:
Is the MCPA genetically modified; if so
provide type of modification
Pythium oligandrum
Species: Pythium oligandrum; genus: Pythium;
family:
Pythiaceae; order: Perensporales; class: Oomycetes.
Pythium oligandrum, M1
Pythium oligandrum is identified using microscopic
taxonomic analysis of species-characteristic spinywalled oospores and oogonia. This method cannot
be used for identification at strain level, but is
sufficient to distinguish Pythium oligandrum from
other Pythium species, and most significantly
Pythium insidiosum.
Pythium oligandrum can be identified at strain level
by a battery of methods including PCR, HPLC MS
MS and multiplication at approprtiate medium. The
PCR on its own has been used to differentiate
between different strains of Pythium oligandrum.
ATCC 38472
Beetween 15 to 20% of the TGAI is used to
manufacture the product.
TGAI conatins approx. 5 % (w/w) of spores of
Pythium oligandrum with remnants of lifeless
mycelium, the number of oospores per g TGAI
ranges from 2.5x 106 – 10 x 106
approx. 95 % (w/w) of fermented remnants of cereal
based growing medium
MPCA is not genetically modified
Biological properties of the micro-organism
.
Origin and natural occurrence, background
level:
Pythium oligandrum is a ubiquitous inhabitant of
the soil where it colonizes the rhizosphere and
rhizoplane of plants and parasitizes other fungi.
Pythium oligandrum has been recorded from soil
and plants in several countries in Europe, Africa,
Australia and North andSouth America.
The background level of Pythium oligandrum Strain
14
Pythium Oligandrum M1
Product-type 10
Target organisms.:
Mode of action:
Host specifity:
Life cycle:
Infectivity, dispersal and colonisation ability:
Relationship to known pathogens:
Genetic stability:
Production of relevant metabolites/toxins:
January 2015
M1 in Danish soil was quantified to be 4 and 26
oospores. g–1 soil (Madsen, 2004). Al-Rawahi
(1997) reported 89-151/g soil.
Saphrophytic moulds occurring on walls and
plastrons
Mycoparasitism, mediated by intimate hyphal
interactions and space competition
Pythium oligandrum is a hyperparazitic micromycet
parasiting more than 20 genera of fungal organisms.
Pythium oligandrum has two cycles of reproduction,
one sexual and one asexual, with the sexual
accounting for around 20 % of the total
reproduction.
Pythium oligandrum is naturally occurring in soil
where is colonizes the rhizosphere of plants. It is
assumed that possible leakage of the microorganism to soil will not increase the levels of
Pythium oligandrum in the environment other than
locally and for a limited time period. Pythium
oligandrum M1 was shown not to be able to grow
on animal tissues and it never grows at 37 O C and
more. Thus infectivity to humans is not expected.
Several Pythium species have been reported to cause
disease in fish and plants (Rivierre et al, 2005). The
only species in the genus that infects mammals is
Pythium insidiosum, known to cause life-threatening
infections in humans and animals. P. insidiosum
differs from all known. Pythium species by
the septation of the main hyphae, as produced in
particular on many agar media, and by the formation
of conspicious, thick-walled fertilization tubes. It
has filamentous, non-inflated sporangia, large
antheridia and high optimum temperature with a
peak growth rate at 37°C and inhibited growth at
40°C.
Stock cultures of lyophilised samples of original
cultures are used for manufacturing of the active
substance, thus preventing genetic drift. In the
available literature there is no indication that
Pythium oligandrum is able to exchange genetic
material with other organisms.
Pythium oligandrum produces several substances
each of which plays a role in the organism’s mode
of action.
These substances include oligandrin, ethylene, cell
wall hydrolytic enzymes such as cellulases,
chitinases,tryphtanime (TNH2) and two types of cell
wall protein fractions, D-type containing two major
proteins with molecular mass of ~28 kDa, and Stype containing one major protein with molecular
mass of ~27 kDa. None ofthese substances are
considered to be of toxicological concern.
As the knowledge about the mode of action of
Pythium oligandrum is still limited, it could be
15
Pythium Oligandrum M1
Product-type 10
Resistance/sensitivity to antibiotics/antimicrobial agents used in human or veterinary
medicine:
Classification and proposed labelling
with regard to micro-organism
January 2015
expected that additional metabolites are produced.
The product is not intended to be mixed with other
fugicides or antibiotics. Pythium oligandrum Strain
M1 is rather sensitive to some fungicides or
antibiotics.
This is supported by Foley & Deacon (1985) who
reported that pentachloronitrobenzene,
chloramphenicol, gallic acid and aureomycin were
moderately or markedly inhibitory to both
mycoparasitic and non-mycoparasitic Pythium spp.
Also, in the applicant´s experience Pythium
oligandrum Strain M1 M1 is sensitive to fungicides
used in agriculture (sulphur, tebuconazol,
chlorthalonil, fenhexamid, mancozeb, folpet,
prothioconazole, fenbuconazole etc.) On the hand,
antibiotics are used in selective agars serving for
isolation of Pythium oligandrum from soils. AlRawahi and Hancock (1997) used for this purpose
semi-selective medium based on cornmeal agar
containing 400 mg of saponin, 25 mg of pimaricin,
50 mg of penicillin G, 50 mg of polymyxin B, 60
mg of streptomycin sulfate, and 20 mg of rose
Bengal per liter. This implies that Pythium
oligandrum Strain M1 is rather resistant against
these antibiotics in the given concentrations.
As a micro-organism Pythium oligandrum
Strain M1 does not fall under the CLP
regulation or Directive 67/548/EE. However,
as other microorganisms it is considered as
a potential sensitizer and its label shall
contain the following phrase:
”Microorganisms may have the potential to
provoke sensitising reactions”
16
Pythium oligandrum M1
Chapter 2:
Product-type 10
January 2015
Methods of Analysis
Analytical methods for the active substance
Manufactured micro-organism(principle of
method):
Impurities and contaminating micro-organisms in
manufactured material (principle of method):
The content of oospores is enumareted under light
microscope in Buerkers chamber.
The following control is performed on the input
inoculum. 1 cm2 of pure culture of Pythium oligandrum
is put in cultivation bouillon, cultivated for 3-4 days at
room temperature and spread on non-selective agar
medium. In case of contamination, the culture of P.
oligandrum excluded from the next technology steps.
The laboratory batches are macroscopically observed
each day of the fermentation process and contaminated
batches are excluded from the production
The same method for quantification of oospores is used
as that used for the active substance.
For each batch, the viability of the oospores is checked
in the following way: 1 g of the preparation is
homogenized in 50 mL of sterile demineralised water and
one drop of Tween 80 is added. The suspension number
of germinating oospores is counted using microscope
after 1, 24, 48, 72, 96 and 120 hours. Enumeration is
carried our four times at each time interval and the
median is calculated.
Analytical methods for residues (viable and non-viable)
Analytical methods for residues of the active micro- The proposed use does not result any significant increase
organism (principle of method):
of M1 strain in any of the environmental comprtments.
Thuspost approval monitoring on a regular basis is not
justified.
In case of accident Pythium oligandrum can be identified
by microscopic taxonomic analysis. Quantifying P.
oligandrum oospores can be made by
diluting the sample and counting the oospores in
Bürker’s chamber. The strain M1 can be differentiated
from other strains using PCR method utilizing
differences in ITS regions and sequences of of
cytochromoxidase genes (Cox I and Cox II) between
differnet Pythium oligandrum Strain M1 strains (
Analytical methods for residues of relevant
metabolites (principle of method):
This method can be complemented by differentiation of
Pythium oligandrum Strain M1 strains from each other
by sequencing the internal transcribed spacer 1 (ITS1,complete sequence, ITS-2(partial sequence) and 5.8S
gene (complete sequence) using FastDNA spin kit. For
determination of the Pythium sequences the primers
described by Schurko and coworkers (2003) and the
protocol described by Allain-Boule and coworkers (2004)
can be utilized.
Pythium oligandrum Strain M1 Oligandrum produces no
toxicologically relevant metabolite . The metabolites
produced by Pythium oligandrum Strain M1 can be
determined by LC MS/MS method combining
digestion of the proteins with trypsin, LC-MS/MS
separation and fragmentation of peptides and
identification of proteins by means of database searching
with obtained MS/MS spectra.
17
Pythium oligandrum M1
Chapter 3:
Product-type 10
January 2015
Impact on Human Health
Medical data, surveillance and observations:
Sensitisation (experience in humans and study
results; type of study):
Acute toxicity
Toxicity after acute oral exposure:
Toxicity after acute inhalation exposure:
Toxicity after acute intraperitoneal/ subcutaneous
exposure:
Infectivity (Annex IIA, point 6.3)
Infectivity after acute oral exposure
Infectivity after acute inhalation exposure
Infectivity after acute intraperitoneal/ subcutaneous
exposure:
Pathogenicity
Pathogenicity after acute oral exposure
Pathogenicity after acute inhalation exposure
Pathogenicity after acute intraperitoneal/
subcutaneous exposure:
No literature reports or any other reports of adverse
effects due to Pythium oligandrum.
No study has been performed.
Pythium oligandrum M1 should be considered as a
potential sensitizer.
LD50 > 5 000 mg kg-1 bw, corresponding to 2.5 x 107
oospores kg-1 bw
LC50 >5 mg/L corresponding to 1 – 1.5 x 104
oospores L-1
LD50 > 3.6 104oospores/animal
Not possible to perform (see Infectiveness study,
below).
Not possible to perform (see Infectiveness study,
below).
Not possible to perform (see Infectiveness study,
below).
Not pathogenic
Not pathogenic
Not pathogenic
Genotoxicity
Pythium oligandrum does not produce any known
metabolites considered to be of toxicological concern.
Genotoxicity testing of specific metabolites
is therefore not required. There are no validated
methods for genotoxicity testing of whole cell
extracts: false positive and false negative results can
both be anticipated and there is a lack of relevant
positive and negative controls, making the results of
any such test difficult to interpret. Until suitable
methods are developed, genotoxicity testing of cell
extracts is not required.
Cell culture study:
Not applicable since Pythium oligandrum is an
oomycete without any known mechanism for
intracellular replication.
Short term toxicity/pathogenicity:
Waived
18
Pythium oligandrum M1
Product-type 10
January 2015
Specific toxicity, pathogenicity and infectiveness studies
Dermal toxicity:
LD50 > 5 000 mg kg-1 bw, corresponding to
approximately 1 x 107 oospores kg-1 bw.
Neurotixicity:
NOAEL = 4g .kg -1 bw day -1 corresponding to 4 x 106
oospores kg -1 bw day -1
Infectiveness study:
Propagules were detected in blood and caecum samples
of all animals at 0 hours after administration. At 1, 3, 6,
and 24 hours, no propagules were found.
Homogenization had a lethal effect on the oospores. On
the basis of the results of this study, performing a
combined toxicity/pathogenicity test on Pythium
oligandrum appeared irrelevant
Growth ability study:
Slight growth of P. oligandrum on potato extract
agar at 26°C and 35°C but no growth at 37°C and
40°C or at any temperature on sheep’s blood agar.
Fructification was not noted at any temperature.
Skin irritation
Not irritating
Eye irritation:
Not irritating
AOEL:
ADI:
Acceptable exposure scenarios
Professional users
Non-professional users
Indirect exposure as a result of use
Not relevant due to lack of significant adverse
effects in relevant studies.
Not relevant due to lack of significant adverse
effects in relevant studies.
Brush and roller application.
Brush and roller application.
Improbable scenario.
19
Pythium oligandrum M1
Product-type 10
January 2015
Chapter 4: Fate and Behaviour in the Environment
Route and rate of degradation in water (Annex IIA, point 7.6, IIIA, point XII.2.1, 2.2)
Hydrolysis of active substance and
relevant metabolites (DT50) (state pH
and temperature)
Not relevant.
Photolytic / photo-oxidative degradation
of active substance and resulting
relevant metabolites
Not relevant
Readily biodegradable (yes/no)
Not relevant
Biodegradation in seawater
Not relevant
Non-extractable residues
Not relevant.
Distribution in water / sediment systems
(active substance)
Not relevant.
Distribution in water / sediment systems
(metabolites)
Not relevant
Route and rate of degradation in soil (Annex IIIA, point VII.4, XII.1.1, XII.1.4;
Annex VI, para. 85)
Mineralization (aerobic)
Not relevant
Laboratory studies (range or median,
with number of measurements, with
regression coefficient)
Not relevant
Field studies (state location, range or
median with number of measurements)
Not relevant
Anaerobic degradation
Not relevant
Soil photolysis
Not relevant
Non-extractable residues
Not relevant
Relevant metabolites - name and/or
code, % of applied a.i. (range and
maximum)
Not relevant
Soil accumulation and plateau
concentration
Not relevant
20
Pythium oligandrum M1
Product-type 10
January 2015
Adsorption/desorption (Annex IIA, point XII.7.7; Annex IIIA, point XII.1.2)
Ka , Kd
Not relevant
Kaoc , Kdoc
Not relevant
pH dependence (yes / no) (if yes type of
Not relevant
dependence)
Fate and behaviour in air (Annex IIIA, point VII.3, VII.5)
Direct photolysis in air
Not relevant
Quantum yield of direct photolysis
Not relevant
Photo-oxidative degradation in air
Not relevant
Volatilization
Not relevant
Monitoring data, if available (Annex VI, para. 44)
Soil (indicate location and type of study)
Not relevant
Surface water (indicate location and type
of study)
Not relevant
Ground water (indicate location and type
of study)
Not relevant
Air (indicate location and type of study)
Not relevant
21
Pythium oligandrum M1
Product-type 10
January 2015
Chapter 5: Effects on Non-target Species
Toxicity data for aquatic species (most sensitive species of each
group)
(Annex IIA, point 8.2, Annex IIIA, point 10.2)
Species
Timescale
Endpoint
Toxicity
Fish
No risk can be
identified with the
proposed use.
Poecillia reticulata
-
Invertebrates
No risk can be
identified with the
proposed use.
Waived
Algae
No risk can be
identified with the
proposed use.
Waived
Microorganisms
Waived
No risk can be
identified with the
proposed use.
Effects on earthworms or other soil non-target organisms
Waived
Acute toxicity to …………………………………..
(Annex IIIA, point XIII.3.2)
Waived
Reproductive toxicity to …………………………
(Annex IIIA, point XIII.3.2)
Effects on soil micro-organisms (Annex IIA, point 7.4)
Nitrogen mineralization
Not relavant.
Carbon mineralization
Not relevant.
Effects on terrestrial vertebrates
Acute toxicity to mammals
(Annex IIIA, point XIII.3.3)
No risk can be identified with the proposed
use.
Acute toxicity to birds
(Annex IIIA, point XIII.1.1)
Waived
Dietary toxicity to birds
Waived
22
Pythium oligandrum M1
Product-type 10
January 2015
(Annex IIIA, point XIII.1.2)
Reproductive toxicity to birds
(Annex IIIA, point XIII.1.3)
Waived
Effects on honeybees (Annex IIIA, point XIII.3.1)
Acute oral toxicity
-
Acute contact toxicity
Indications that there might not be any
effects with approximately 103 oospores per
bee.
Effects on other beneficial arthropods (Annex IIIA, point XIII.3.1)
Acute oral toxicity
-
Acute contact toxicity
The indicative test on bees can also be valid
for other arthropods, in which there is an
indication of no effect at an exposure rate of
approximately 103 oospores per bee, which
means that one bee need to be sprayed with
1 l.
-
Acute toxicity to …………………………………..
Bioconcentration (Annex IIA, point 7.5)
Bioconcentration factor (BCF)
Not relevant.
Depration time
Not relevant.
(DT50)
(DT90)
Level of metabolites (%) in organisms
accounting for > 10 % of residues
Not relevant.
23
Pythium oligandrum M1
Product-type 10
January 2015
Appendix II: List of Intended Uses
Object
and/or
situation
Product
name
Organisms
controlled
Formulation
Type
(d-f)
Conc.
of a.s.
(i)
Application
method
kind
(f-h)
number
min max
Applied amount per treatment
interval
between
applications
(min)
g a.s./L
min max
water
L/m2
min
max
Wall
(curative)
Wall
Biorepell
(preventive,
paint)
powder
20%
brushing
2
2
2 hours
0.02*
0.2* 1.5*
g a.s./m2 *two
min max treatments
with different
doses and
concentrations
performed
0.04
molds
Biorepell
(preventive)
Wall
Saprophytic
Saprophytic
powder
20%
brushing
1
1
n.a.
0.04 – 0.1
molds
Biorepell
Saprophytic
powder
20%
brushing
1
1
n.a.
0.12 – 0.12
10*
0.04
0.15
0.006
0.5
0.05
0.072
0.00864
0.1
0.012
molds
24
Re
marks:
Pythium Oligandrum
Product-type 10
January 2015
Appendix III: List of studies
Data protection is claimed by the applicant in accordance with Article 60 of Regulation (EU)
No 528/2012.
Section
Author(s)[2] Year Title[3]
Data
No
/
Source Company Protection
Reference
Report
No. Claimed
No[1]
GLP
(where (Yes/No)
relevant)
Owner
(Un)Published
IIA 2.1
IIIA 1.3.3
Jong SC, &
De Lucio, A.
1978 Re: Phytopath. Z. 90: 113115, 1977 Pythium
oligandrum a strain for
biological control of
damping off pathogens
YES
Biopreparáty,
spol. s r.o.
YES
Biopreparáty,
spol. s r.o
NO
Public
NO
Public
GLP: No
Published: No
IIA 2.1
IIIA 1.3.3
Laichmanová M
2005 Záznam o deponování
kultury za účelem
bezpečného uložení, Czech
Collection of Microorganisms
GLP: No
IIA 2.3
IIA 2.4.3
IIA 5.1
IIIA 1.3.
IIIA 2.5
IIIA 2.6
IIIA 2.10
IIIA 4.1
IIA 2.4.3
IIIA 1.3.4
IIIA 4.1
Van der PlaatsNiterink J
Published: No
1981 MONOGRAPH OF THE
GENUS PYTHIUM , Studies
in Mycology, 21: 1-242
GLP: No
Published: Yes
Godfrey SAC,
Monds, RD,
Lash DT
and Marshall JW
2003 Identification of
Pythium oligandrum
using species-specific
ITS rDNA PCR
oligonucleotides.
Mycological
research 107(7):790-
[1]
Section Number/Reference Number should refer to the section number in Doc III-A or III-B. If the study is non-key, and
hence not summarised in Doc III but mentioned in Doc II, it should be included in the reference list alongside related references
and its location in Doc II indicated in brackets. (If there is a need to include a cross-reference to PPP references then an
additional column can be inserted).
[2]
Author’s Name should include the author’s surname before initial (s) to enable the column to be sorted alphabetically. If the
Human Rights Charter prevents author’s surnames on unpublished references being included in non-confidential documents, then
it will be necessary to consider including ‘Unpublished [number/year & letter] ’ in Doc II, and both ‘ Unpublished [number/year
& letter]’ and the ‘Authors Name’ in the reference list’. This may necessitate the need for an additional column to state whether a
reference is unpublished which can then be sorted.
[3]
Title, Source (where different from company), Company, Report No., GLP (where relevant), (Un)Published should contain
information relevant to each item (ideally on separate lines within the table cell for clarity). If useful, the name of the electronic
file containing the specific study/reference could be added in brackets.
25
Pythium Oligandrum
Product-type 10
January 2015
Section
Author(s)[2] Year Title[3]
Data
No
/
Source Company Protection
Reference
Report
No. Claimed
[1]
No
GLP
(where (Yes/No)
relevant)
(Un)Published
796.
GLP: No
Published: Yes
IIIA 1.4.3
2002
YES
IIIA 1.4.3
2006
YES
IIIA 1.4.3
2014
YES
IIIA 1.4.3
2014a
YES
26
Owner
Pythium Oligandrum
Product-type 10
January 2015
Section
Author(s)[2] Year Title[3]
Data
No
/
Source Company Protection
Reference
Report
No. Claimed
[1]
No
GLP
(where (Yes/No)
relevant)
(Un)Published
IIIA 1.4.3
2014b
YES
IIIA 1.4.3
2014c
YES
IIIA 1.4.3
2014d
YES
IIIA 1.4.3
2014e
YES
27
Owner
Pythium Oligandrum
Product-type 10
January 2015
Section
Author(s)[2] Year Title[3]
Data
No
/
Source Company Protection
Reference
Report
No. Claimed
[1]
No
GLP
(where (Yes/No)
relevant)
Owner
(Un)Published
IIIA 2.1.1
Martin FN, &
Hancock JG
1987 THE USE OF PYTHIUM
OLIGANDRUM FOR
BIOLOGICAL
CONTROL OF
PREEMERGENCE
DAMPING-OFF
CAUSED BY PYTHIUM
ULTIMUM,
Phytopathology, 87
(7): 1013-1020
NO
Public
NO
Public
NO
Public
NO
Public
NO
Public
GLP: No
Published: Yes
IIIA 2.1.1
Veselý D
1977 POTENTIAL
BIOLOGICAL
CONTROL OF
DAMPING-OFF
PATHOGENS IN
EMERGING SUGAR
BEETS BY PYTHIUM
OLIGANDRUM
DRECHSLER.
Phytopath. Z. 90,
113-115 GLP: No
Published: Yes
IIA 2.3
IIA 6
IIIA 2.2.2
Laing SAK, &
Deacon JW
1991 Video microscopical
comparison of
mycoparasitism by Pythium
oligandrum, P. nunn, and
an unnamed Pythium
species, Mycological
Research 95 (4): 469-479.
GLP: No
Published: Yes
IIA 2.3
IIA 5.1
IIA 5.2.4
IIA 6
IIIA 2.2.2
Lewis K, Whipps
JM, & Cook RC
1989 Mechanisms of biological
disease control with special
reference to the case study
of Pythium oligandrum as
an antagonist. In:
Biotechnology of Fungi for
Improving Plant Growth.
British Mycological Society.
GLP: No
Published: Yes
IIA 2.3
IIIA 2.3
Brožová J
2002 Exploitation of the
mycoparasitic fungus
Pythium oligandrum in
plant protection. Plant
28
Pythium Oligandrum
Product-type 10
January 2015
Author(s)c21 Year TitleC3 l
Section
Data
No
Source Company Protection
I
Reference
Report
No. Claimed
NoC1 l
(where
GLP
(Yes/No)
relevant)
Owner
(Un)Published
Protection Science. 38: 2935
GLP:No
Published: Yes
IIA 2.3
Deacon J
2006 Fungal biology, 4th edition
NO
Public
NO
Public
Blackwell Publishing Ltd
GLP:No
Published: Yes
IIA 2.3
Kwon-Chung
KJ
IIA 2 .3
IIA 5 .1
IIA 5 .2.4
llA 2.3
1994 Phylogenetic
Spectrum of Fungi
That Are Pathogenic
to Humans, Clinical
Infectious Diseases,
19 (Suppl 1) : 1-7.
Rivierre C,
2005
laprie C,
IIIA 2. 3
YES
2006
PYTHIOSIS IN AFR ICA,
Guiard-Marigny
Diseases, 11: 479-481
O, Bergeaud P,
GLP:No
Berthelemy M, &
Gui llot J
Published: Yes
Fletcher JT,
NO
Public
NO
Public
Emerging lnfectuos
1990 PYTHJUM OLIGANORUM
ASSOCIATED WITH A
Smewin BJ, &
O'Brien A
CROPPING DISORDER OF
AGAR/CVS BJSPORUS. Plant
Pathology, 39, 603-605.
GLP: No
IIIA 2 .4
IIA 2.4.2
-
Published: Yes
2003
YES
2013
YES
29
Pythium Oligandrum
Product-type 10
January 2015
Author( s) £2 l Year TitleC3 l
Section
Data
Source Company Protection
No
I
Reference
Report
No. Claimed
No£1 l
GLP
(where (Yes/No)
relevant)
(Un)Published
IIA 2.4.2
2013a
YES
IIA 2.4.3
2013b
YES
IIA 2.4.3
2013
YES
IIA 2.4.3
2013a
YES
30
Owner
Pythium Oligandrum
Product-type 10
January 2015
Section
Author(s)[2] Year Title[3]
Data
No
/
Source Company Protection
Reference
Report
No. Claimed
[1]
No
GLP
(where (Yes/No)
relevant)
Owner
(Un)Published
IIA 2.4.3
Vallance J
2009
Influence of Pythium
oligandrum Biocontrol
on Fungal and
Oomycete Population
Dynamics in the
Rhizosphere
NO
Public
NO
Public
NO
Public
NO
Public
GLP: No
Published: Yes
IIA 2.4.3
Schroeder KL,
Martin FN,
de Cock
AWAM,
Lévesque CA,
Spies CFJ,
Okubara PA, &
Paulitz TC
2013
Molecular Detection
and Quantification of
Pythium Species:
Evolving Taxonomy,
New Tools, and
Challenges. Plant
Disease, January
2013, Volume 97,
Number 1 Pages 4 –
20
GLP: No
IIA 2.4.3
Lévesque CA,
& de Cock
AWAM
,
2003
Published: Yes
Molecular phylogeny
and taxonomy of the
genus Pythium.
Mycol. Res.
108:1363–1383
GLP: No
Published: Yes
Schurko AM,
Mendoza L,
Lévesque CA,
Désaulniers
NL,
De Cock
AWAM, &
Klassen GR
2003
A molecular
phylogeny of
Pythium, Mycol. Res.
107:537–544
GLP: No
Published: Yes
31
Pythium Oligandrum
Product-type 10
January 2015
Section
Author(s)[2] Year Title[3]
Data
No
/
Source Company Protection
Reference
Report
No. Claimed
[1]
No
GLP
(where (Yes/No)
relevant)
Owner
(Un)Published
IIA 2.4.3
Allain-Boulé
N, Tweddell R,
Mazzola M,
Bélanger R, &
Lévesque CA
2004
Pythium
attrantheridium sp.
nov.: taxonomy and
comparison with
related species.
Mycol. Res. 108:795–
805
NO
Public
NO
Public
NO
Public
NO
Public
NO
Public
YES
Biopreparáty,
GLP: No
Published: Yes
IIIA 2.6
Mendoza L,
Hernandez F, &
Ajello L
1993
Life Cycle of the Human
and Animal Oomycete
Pathogen Pythium
insidiosum, Journal of
Clinical Microbiology, vol.
31, no 11: 2967-73.
GLP: No
Published: Yes
IIIA 2.8
IIIA 2.8
IIIA 2.8
IIA 4.2
IIIA
Picard K, Ponchet
M, Blein JP, Tirilly
Y, & Benhamou N
2000
Hase S, Shimizu A,
Nakaho K,
Takenaka S, &
Takahashi H
2006
Le Floch G, Rey P,
Benizri E,
Benhamou N, &
Trilly Y
2003
1979
Oligandrin. A
Proteinaceous Molecule
Produced by the
Mycoparasite Pythium
oligandrum Induces
Resistance to Phytophthora
parasitica Infection in
Tomato Plants, Plant
Physiology 124 (1): 379395.
GLP: No
Published: Yes
Induction of transient
ethylene and reduction in
severity of tomato bacterial
wilt by Pythium
oligandrum, Plant
Pathology, 55, 537–543
GLP: No
Published: Yes
Impact of auxincompounds produced by
the antagonistic fungus
Pythium oligandrum or the
minor pathogen Pythium
group F on plant growth,
Plant and soil, 104 (1): 1109.
GLP: No
Published: Yes
Realisation of the
experiment – acute toxicity
32
Pythium Oligandrum
Product-type 10
January 2015
Author(s)c21 Year TitleC3 l
Section
Data
No
Source Company Protection
I
Reference
Report
No. Claimed
NoC1 l
(where
GLP
(Yes/No)
relevant)
Owner
(Un)Published
5. 2.2 .1
IIA 4.2
IIIA
5. 2.2.1
IIA 4 .2
IIIA
5. 2.2.1
spol. s r.o
GLP: No
-
Published: No
1980
Realisation of the
experiment - acute t oxicity
YES
Biopreparat y,
spol. s r.o
of given preparat ion
GLP:No
Published: No
2001
Oospores of fungic
YES
organism Pythium
o/igandrum Drechsler -
Biopreparat y,
spol. s r.o
Acute Oral Toxicity in Rats
Published: No
IIA 4.2
IIIA
5. 2.2 .1
2001
Oospores of f ungic
organism Pythium
YES
Biopreparaty,
spol. s r.o
YES
Biopreparat y,
spol. s r.o
YES
Biopreparat y,
spol. s r.o
YES
Biopreparat y,
spol. s r.o
o/igandrum Drechsler -
GLP: Yes
Published: No
IIA 4.2 .2
IIIA
5. 2.2.2
2002
Concentrat e of Pythium
o/igandrum Drechsler
oospores inclu ding ot her
propagules - Acute
Inhalation Toxicity - Lli mit
GLP: Yes
Published: No
I
IIA 4 .2.3
IIIA
5. 2 .2 .3
IIA 4.3. 1
IIIA
5. 2.2 .3
I-
2005
Propagules Pythium
o/igandrum DV 74 - Acute
GLP: Yes
Published: No
12003
Concentrate of Pythium
o/igandrum Drechsler
oospores inclu ding ot her
33
Pythium Oligandrum
Product-type 10
January 2015
Author( s) £2 l Year TitleC3 l
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No£1 l
(where
GLP
(Yes/No)
relevant)
Owner
(Un)Published
propagules Subcut aneous
Dermal Tolerance - Rabbit
IIA 4. 3 .1
YES
2005
Biopreparaty,
spol. s r.o
GLP: Yes
Published: No
IIA 4.2 .1
IIIA
5. 2.2 .4
2001
Concentrate of Pythium
o/igandrum Drechsler
YES
Biopreparaty,
spol. s r.o
oospores i ncluding other
propagules - Dermal Acut e
GLP: Yes
Published: No
IIIA
5. 2.2.4
2002 Concentrate of Pythium
o/igandrum Drechsler
YES
Biopreparaty,
spol. s r.o
GLP: Yes
Published: No
IIA 4 .5
IIIA 5.2 .5
1981
Realisation of experiment sub-acut e t oxicity of
biologica l preparation for
YES
Biopreparaty,
spol. s r.o
GLP: No
Published: No
IIA 5 .1
IIIA 7 .1
Foley, MF, &
Deacon, JW
1985 Isolation of Pythium
o/igandrum and other
necrotrophic mycoparasites
from soil. Trans.Br. Mycol.
Soc. 84(4):631-639
GLP: No
34
NO
Public
Pythium Oligandrum
Product-type 10
January 2015
Author(s)c21 Year TitleC3 l
Section
Data
No
Source Company Protection
I
Reference
Report
No. Claimed
NoC1 l
(where
GLP
(Yes/No)
relevant)
Owner
(Un)Published
Published: Yes
IIA 5 .1
IIIA 7 .1
2013a Horizontal mobility of
Pythium oligandrum
in defined soil
environment.
YES
IIIA 7 .1
2013b Behaviour of Pythium
oligandrum on
interface of
sedimentary phase
and water
environment
YES
Biopreparaty,
spol. s r.o.
NO
Public
NO
Public
NO
Public
IIA 5 .1
Al- Rawah i
AK,
& Hancock JG
1997
Rhizosphere
Competence of
Pythium oligandrum.
Phytopathology
87 : 951-959.
Biopreparaty,
spol. s r.o.
GLP: No
Published: Yes
IIA 5 .1.2
Madsen MA, & de
Neergaard E
2004
Interactions Between the
Mycoparasite Pythium
o/igandrum and Sclerotia of
the Plant Pathogen
Sclerotinia sclerotiorum .
European Journal of Plant
IIA 5.1. 2
Takenaka S,
2008
Pathology, Vol. 105, Issue 8,
761-768).
Colonization of Pythium
o/igandrum in the Tomato
Sekiguchi H,
Nakaho K, Tojo
Rh izosphere for Biological
Control of Bacterial Wi lt
Disease Analyzed by Rea lTime PCR and Confocal
M, Masunaka A,
& Takahash i H
Laser-Scanning Microscopy
IIIA 8 .2 .1
-
Phytopathology. 2008,
2003
Feb;98{2):187-95
Concentrate of Pythium
o/igandrum Drechsler
oospores including other
propagules - Test of Acute
Toxicity (Poeci/ia reticu/ata)
35
YES
Biopreparaty,
spol. s r.o.
Pythium Oligandrum
Product-type 10
Author( s) £2 l Year TitleC3 l
Section
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No
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Reference
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No. Claimed
No£1 l
(where
GLP
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relevant)
January 2015
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(Un)Published
GLP : YES
Published : No
IIIA 8 .3
VeselyV
1992
Classification of the
YES
preparation accord ing to its
effect on bees - bind ing
expertise, Extract from the
protocol No.330 BeeKeeping Research Institute,
Doi, Laboratory report: 330
No. of decision 3953/92,
report date: 11th and 12'h
December 1992
GLP: No
Published : No
36
Biopreparaty,
spol. s r.o.