epa staff evaluation and review report

EPA STAFF EVALUATION AND REVIEW REPORT

www.epa.govt.nz

Application for approval to import and manufacture Kasumin for

release

2

Application for approval to import and manufacture Kasumin for release (APP201581)

September 2013

Executive Summary

Background information

1. ETEC Crop Solutions Limited has applied for approval to import or manufacture Kasumin containing

kasugamycin as the active ingredient. Kasumin is intended for use as a bactericide antibiotic to control

Pseudomonas syringe pv. actinidiae (Psa) in kiwifruit.

2. Kasumin is a water soluble concentrate containing 20 g/L kasugamycin. It will be diluted with water and

applied as a high-volume ground-based spray to kiwifruit vines, foliage and stems.

3. Staff consider that this substance has a high level of benefits, providing a new tool for the control of Psa.

Psa has had a serious economic impact on kiwifruit production in New Zealand and any new

management tools are warmly welcomed by the industry.

Classification

4. The staff have classified Kasumin based on its composition and the effects of its components:

Hazard Endpoint HSNO classification

Acute toxicity 6.1E (dermal, inhalation)

Reproductive/developmental toxicity 6.8B

Target organ systemic toxicity 6.9B

Submissions

5. In response to public notification of the application, four submissions were received. One opposed the

application, one supported it, and two neither opposed nor supported but had concerns about some

parts of the application.

6. The main concern highlighted in the submissions is the risk that antibiotic residues pose to the bee-

product export market. The staff consider that this risk will be more appropriately assessed by ACVM

during their registration process as they have jurisdiction over the control of the quality of exports for

New Zealand.

7. Two submitters requested to be heard at a public hearing.

Risk and benefits assessment

8. The greatest concern identified during the risk assessment was uncertainty regarding the completeness

of the data package provided. In general, the staff have been able to accept the conclusions of other

regulators, in combination with study summaries to make risk assessments when the full studies were

not made available by the applicant. There are however, several outstanding data gaps which are, in the

opinion of the staff, significant and should be mitigated. Specifically:

Effects of kasugamycinic acid on aquatic organisms (fish, invertebrates, algae).

Effects of kasugamycin on the reproduction of earthworms.

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Application for approval to import or manufacture Kasumin for release (APP201581)

September 2013

Effects of kasugamycin on the carbon transformation in soil.

Effects of kasugamycin on bee larvae.

Higher tier studies on non-target arthropods with kasugamycin.

Data on dermal absorption and/or dislodgeable foliar residues and foliar half life

9. The risks to re-entry workers are identified as high, and so a re-entry interval has been proposed.

10. The risks to bystanders are acceptable provided that a control is put in place to mitigate spray-drift, given

the use of an airblast sprayer in a sparse orchard.

11. The risks to aquatic organisms, earthworms, non-target plants, bees and birds are considered low.

12. Kasumin presents a high risk to non-target arthropods in-field and a possible risk to them off-field as

well.

13. Benefits resulting from the availability and use of Kasumin will include:

The use of a new antibiotic ingredient that does not have additional uses as a human or veterinary

medicine, thus avoiding any issues with antibiotic resistance;

Reducing the economic damage caused by Psa to the kiwifruit industry; and

Replacing more harmful current control methods (e.g. streptomycin-based products).

Controls

Default controls

14. A number of default controls are prescribed by regulations under the Hazardous Substances and New

Organisms Act 1996 (the Act) as a consequence of the toxicity classifications of this substance. These

controls form the basis of the controls that are listed in Appendix C.

15. The staff consider that it is appropriate for certain variations to be made to the default controls. These

variations are discussed in Section 7 of this Evaluation and Review (E&R) Report and listed in Appendix

C.

Additional controls

16. The staff propose the addition of a number of controls for the substance under section 77A of the Act.

17. The label must contain the following statement:

“Kasumin contains the antibiotic, kasugamycin, and may be harmful to beneficial organisms, such as

predatory mites, earthworms, and bees and other pollinators.”

18. The use of Kasumin should be restricted so that:

The substance shall not be applied into, onto or over water.1

1 Where ‘water‘ means water in all its physical forms, whether flowing or not, but does not include water in any form while in a pipe, tank or cistern or water used in the dilution of the substance prior to application

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September 2013

19. The application of the substance should be limited in the following way:

Maximum application rate 100 g/ha, 4 applications maximum per season with a minimum interval of 10

days.

Kasumin must only be applied via ground-based methods.

20. An approved handler requirement should be imposed, to help mitigate the level of potential risk to the

environment.

21. A restricted entry interval should also be put in place to protect bystanders.

Conclusion

22. The staff’s risk assessment indicates that with controls in place, there is a negligible level of risk to

human health and non-negligible level of risk to the environment from the use of Kasumin.

23. Additional controls have been proposed to mitigate these non-negligible environmental risks. Even with

the additional controls in place though, the uncertainty around several endpoints means that there will

still be non-negligible risks associated with the use of Kasumin.

24. Given the high level of benefits associated with using Kasumin to control Psa in kiwifruit, it is the staff’s

opinion that a determination under clause 27 of the Methodology is appropriate.

25. With the proposed controls in place, the overall level of benefit provided by the availability of Kasumin

(high) is greater than the level of adverse effects (low).

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September 2013

Table of Contents

Executive Summary ............................................................................................................................... 2

Table of Contents .................................................................................................................................. 5

1. Summary ...................................................................................................................................... 6

2. Background .................................................................................................................................. 6

3. Process, consultation and notification ..................................................................................... 7

4. Hazardous properties ................................................................................................................. 8

5. Submissions ................................................................................................................................ 8

6. Risk, cost and benefit assessment............................................................................................ 9

7. Controls ...................................................................................................................................... 16

8. Overall evaluation and recommendation ................................................................................ 19

Appendix A: Staff classification of Kasumin .................................................................................... 20

Appendix B: Risk assessment ......................................................................................................... 119

Appendix C: Controls applying to Kasumin ................................................................................... 141

Appendix D: References ................................................................................................................... 147

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September 2013

1. Summary

Application Code APP201581

Application Type To import or manufacture for release any hazardous substance under Section

28 of the Hazardous Substances and New Organisms Act 1996 (“the Act”)

Application Sub-Type Notified - Category C

Applicant ETEC Crop Solutions Limited

Purpose of the application To import Kasumin bactericide antibiotic containing kasugamycin for the control

of Pseudomonas syringe pv. actinidiae (Psa-V) in kiwifruit.

Date Application Received 28 May 2013

Submission Period 12 June 2013 – 24 July 2013

Submissions received Four submissions were received

Information request The consideration of the application was postponed to allow further data to be

provided by the applicant and assessed by the staff

2. Background

2.1. Kasumin is a bactericidal antibiotic containing 20 g/L of kasugamycin. It is in the form of a water

soluble concentrate. Kasumin is intended for use as a bactericide for the control of Pseudomonas

syringe pv. actinidiae (Psa-V) in kiwifruit.

2.2. The applicant has described the lifecycle of the substance as follows.

Transport and storage

2.3. The substance is manufactured in Japan and is formulated as a water soluble concentrate and packed

and stored in 180 litre drums. It will be repacked into labelled polyethylene (PE) containers of capacity

1-20 litres for retail distribution as marketable packs. Transport to New Zealand is via sea or air. The

substance is imported into New Zealand by the sole New Zealand agent, fully formulated and

packaged and transported to a designated depot for storage of chemical products for eventual delivery

to dealers and retail distributor premises. Storage will be in an Auckland warehouse contracted for this

service. Transport throughout New Zealand of bulk cartons containing multiples of these packs would

be by licensed road carriers, whilst transport in public passenger vehicles is not envisaged and

unlikely.

Use

2.4. The substance is to be used specifically to aid in the control of the bacterial disease Pseudomonas

syringae pv. actinidiae (Psa-V) in kiwifruit. Application is made by dilution in water and applied as a

high volume ground based spray to the vine foliage and stems. Applicator equipment would include

calibrated commercial orchard sprayers.

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September 2013

2.5. The users are expected to be commercial contractors and kiwifruit growers who are approved

handlers for storage, handling and use of hazardous pesticides.

Disposal

2.6. The plastic container is retained until emptied of the liquid contents. Any residual substance is diluted

with water and disposed of onto waste barren ground or added to the rinsate in the spray tank. The

empty container is not refilled, reused or burnt but can be disposed of by recycling in an appropriate

program (e.g. AgRecovery). The empty container is triple rinsed and rinsate added to the spray tank.

An alternate disposal method of the empty container is according to the local council by-laws for

industrial waste or burial in a suitable landfill.

3. Process, consultation and notification

3.1. The application was lodged pursuant to section 28 of the Act.

3.2. The Labour Group of the Ministry of Business, Innovation and Employment (MBIE)2, the Agricultural

Compounds and Veterinary Medicines (ACVM) Group of the Ministry of Primary Industries, the

Ministry of Health and the Department of Conservation (DOC) were advised of the application. No

comments were received.

3.3. The application was publicly notified as it was considered that there was likely to be significant public

interest in the application because it contains an active ingredient which has not previously been

approved in New Zealand. It was also considered that there might be significant interest in Kasumin

because of its intended use.

3.4. Four submissions were received. One opposed the application, one supported it, and two did not

indicate opposition or support, but had concerns about some parts of the application.

3.5. Initially, the application did not contain sufficient information for the staff of the EPA (“the staff”) to

undertake a full assessment of the substance from a scientific and technical perspective. The staff

requested that a number of studies be provided as further information under section 58 of the Act.

3.6. In response to this information request, the applicant has provided most of the requested studies to

the EPA. However, some of the requested studies are still not available, and the staff consider that

they have not received sufficient information from the applicant to assess all of the risks associated

with the release of Kasumin. As a result, the staff consider there is significant uncertainty regarding

the assessment of the risks associated with application of Kasumin in the manner proposed by the

applicant.

3.7. In preparing this report, the staff took into account:

The application form;

The submissions received;

2 Formerly the Department of Labour.

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September 2013

Additional information received from the applicant;

The response to the submissions from the applicant; and

Existing approvals for other antibiotics like streptomycin.

4. Hazardous properties

4.1. The staff determined the hazard profile of Kasumin based on the information provided by the applicant

and other available information as documented in Appendix A.

Table 1: Hazard classification of Kasumin

Hazard Endpoint Applicant Staff

Acute toxicity - 6.1E (dermal, inhalation)

Eye irritancy 6.4A No

Reproductive/developmental

toxicity - 6.8B

Target organ systemic toxicity - 6.9B

Aquatic toxicity 9.1C No

5. Submissions

5.1. This application was open for submissions from 12 June 2013 to 24 July 2013. Four submissions were

received. The submissions are available on the EPA website.

5.2. The main concerns identified by submitters were around the risk that antibiotic residues pose to the

bee-product export market. The staff consider this is an adverse effect which is better considered

during ACVM registration. This is because it relates to the safety of exported food and food-related

products which is covered under the Animal Products Act (1999). The Ministry for Primary Industries

(MPI) are also tasked with considering risks to trade under the Agricultural Compounds and Veterinary

Medicines (ACVM) Act (1997).

5.3. Concerns were also raised by submitters about the lack of proven benefits of the substance. It is

difficult to accurately estimate the magnitude of a benefit prior to distribution through the marketplace,

and so the staff assessment must carefully consider the likelihood of a benefit arising. This is in part

due to the fact that evidence of efficacy testing is usually submitted as part of an ACVM registration,

but not with a HSNO application. The staff consider it is highly likely that some magnitude of benefit

will be realised through the use of this substance. The staff also note the submission from David

Tanner (on behalf of Zespri, Kiwifruit Vine Health and New Zealand Kiwifruit Growers Incorporated)

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September 2013

which demonstrates that the kiwifruit industry consider Kasumin is likely to be effective in the fight

against Psa and thus has a high level of benefits.

5.4. Submitters also raised concerns that there is insufficient information provided to assess the risk to

bees. The staff share this concern and a study on the effects of kasugamycin on bee larvae has been

requested, but not provided to date.

6. Risk, cost and benefit assessment

6.1. The staff’s identification and assessment of risks and costs (adverse effects) and benefits (positive

effects) is set out in this section and supported by information in Appendix B.

Risks and costs

Human health

6.2. The staff have evaluated the potential of kasugamycin and Kasumin to cause adverse effects to the

health and safety of humans during all stages of the substance’s lifecycle using qualitative risk

assessment methodology, and qualitative assessment to determine risks associated with use of the

substance, entry into treated areas and effects on bystanders.

6.3. The staff have classified Kasumin as acutely toxic (6.1E), a reproductive / developmental toxicant

(6.8B), and a target organ toxicant (6.9B).

6.4. In the staff’s opinion, chronic hazards normally require repeated exposure to the substance for the

adverse effects to occur and are therefore most relevant to the end-users.

6.5. The risks of Kasumin to human health and safety at various stages of its lifecycle are summarised in

Table 2.

Environmental

6.6. The staff have evaluated the potential of Kasumin to cause adverse effects to the environment during

all stages of the substance’s lifecycle using a qualitative risk assessment methodology, and qualitative

assessment to determine risks associated with use of the substance.

6.7. The staff have not classified Kasumin as a class 9 substance.

6.8. The risks of Kasumin to the environment at various stages of its lifecycle are summarised in Table 3.

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September 2013

Table 2: Assessment of risks to human health for Kasumin

Lifecycle Description Likelihood Magnitude Matrix Comment Level of risk

Manufacture3

and

packaging

Acute toxicity

(dermal, inhalation)

Highly

improbable

Minimal Negligible Manufacturing and packaging facilities in New Zealand will

be required to meet the HSNO requirements for equipment,

emergency management and Personal Protective

Equipment (PPE). Compliance with HSNO information

provisions (e.g. labels, advertising, Safety Data Sheets

(SDS), and MBIE Health and Safety requirements will also

apply.

Negligible

Reproductive /

developmental

toxicity

Highly

improbable

Major Low While the qualitative descriptors indicate a low level of risk

driven by the major chronic effects, the staff consider that

these requirements will make the likelihood of exposure

that would lead to a chronic effect so highly improbable that

the level of risk for the chronic toxic adverse effects is

negligible.

Negligible

Target organ

toxicity

Highly

improbable

Major Low Negligible

Importation,

transport,

storage

Acute toxicity


Highly

improbable

Minimal Negligible Workers and bystanders will only be exposed to the

substance during this part of the lifecycle in isolated

incidents where spillage occurs. HSNO controls (e.g.

labels, SDS and packaging) and adherence to the Land

Transport Rule 45001, Civil Aviation Act 1990 and Maritime

Transport Act 1994 (as applicable) will apply.

Negligible

3 The applicant intends to import (not manufacture) Kasumin. However, it is possible that the substance could be manufactured in New Zealand in the future. Consequently,

the risks associated with the manufacture of Kasumin have been evaluated so the approval of this substance will be applicable to both the import and manufacture Kasumin.

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September 2013


Reproductive /

developmental

toxicity

The risk of chronic effects from exposure during importation, transport and storage is sufficiently remote that

the level of risk will be negligible.

Negligible

Target organ

toxicity

Negligible

Use

Acute toxicity


Highly

improbable

Moderate Negligible The substance will be labelled to identify its potential risks

minimising the opportunity for it to cause toxicity. HSNO

requirements for PPE, packaging, identification and

emergency management requirements must be complied

with. Compliance with the controls requiring users to wear

appropriate PPE will protect users from accidental

inhalation of or dermal exposure to the substance.

Negligible

Reproductive /

developmental

toxicity

The risks to operators are acceptable provided workers (operators) use full PPE with or without a respirator as

appropriate during mixing, loading and application.

The risks to re-entry workers are identified as high. The staff consider that the estimates are likely to

substantially over estimate re-entry risks, primarily due to the use of the default 50% dermal absorption value

(in the absence of specific dermal absorption data) and a lack of information on dislodgeable foliar residues and

foliar half-life. However, in the absence of specific data it is recommended that a re-entry interval is established

for as long a duration as is reasonably practicable (up to 30 days with gloves or 40 days with no PPE).

The risks to bystanders are acceptable, given controls imposed to protect the environment will prevent the use

of aerial application. If application is not restricted to ground-based methods only, then the staff would

recommend a restriction on spray droplet size.

While the RQ value for bystanders from airblast application in a sparse orchard is slightly above the level of

concern, this is likely to be an over estimate and it is therefore considered that this use scenario is unlikely to

present a significant risk of adverse effects.

Negligible

Target organ

toxicity

Negligible

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September 2013


Disposal

Acute toxicity


Highly

improbable

Minor Negligible The applicant indicates that the rinse water is to be added

to the spray tank. The triple-rinsed empty containers or

unused product would also be suitable for the collection by

the AgRecovery container and chemical recycling

programme.

In all cases of disposal, the substance will be disposed of in

accordance with the requirements of the Hazardous

Substances (Disposal) Regulations 2001 and the Resource

Management Act 1991.

Negligible

Reproductive /

developmental

toxicity

The risk of chronic effects from exposure during disposal is sufficiently remote that the level of risk will be

negligible.

Negligible

Target organ

toxicity

Negligible

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September 2013

Table 3: Assessment of risks to the environment for Kasumin


Manufacture4,

importation,

transport and

storage

Death or

adverse effects

to aquatic or

terrestrial

organisms

Highly

improbable

Moderate Negligible Provided there is adherence to the HSNO controls (and the Land

Transport Rule 45001, the Civil Aviation Act 1990 and the

Maritime Transport Act 1994 (as applicable)) the staff consider a

spill to be highly improbable. Furthermore, provided importers,

manufacturers and those handling the substances adhere to the

HSNO controls relating to storage and bunding requirements, a

spill is only likely to lead to localised effects.

Negligible

Use

Death or

adverse effects

to aquatic

organisms

Formulation data have been used for the risk assessment. Low acute and chronic risks have been identified for

aquatic organisms from the use of Kasumin in kiwifruit. However, it was not possible to evaluate the risk from the

significant metabolite (kasugamycinic acid) due to data lacking.

Low

Death or

adverse effects

to earthworms

Low acute risks have been identified to earthworms from the use of Kasumin in kiwifruit, from in-field as well as off-

field exposures. No data are available about the chronic toxicity of kasugamycin on earthworms so no risk

assessment was performed.

Low

Death or

adverse effects

to non-target

plants

The studies on vegetative vigour and seedling emergence of 10 species of non-target plants show that the maximum

application rate does not produce more than 50% of effects. Therefore no risk assessment is necessary and the risk

is considered to be low according to the Guidance Document on terrestrial ecotoxicology Table . The Canadian

authorities concluded that a buffer zone of 2 m was necessary to protect the non-target plants however, their

assessment is based on ER25 values, which is not consistent with our current approach.

Negligible

Death or

adverse effects

to terrestrial

vertebrates

No mortality was observed in the 3 acute tests and the 2 short-term tests on birds. In addition the NOEC from the 2

chronic tests was the highest tested concentration (1000 ppm) so no risk assessment for birds was performed due to

the low toxicity of kasugamycin. The risks are considered to be very low.

Negligible

4 The applicant intends to import (not manufacture) Kasumin. However, it is possible that the substance could be manufactured in New Zealand in the future. Consequently, the risks associated with the manufacture of Kasumin have been evaluated so the approval of this substance will be applicable to both the import and manufacture Kasumin.

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September 2013


Death or

adverse effects

to terrestrial

invertebrates

The HQoral value is below the trigger value of 50, indicating that kasugamycin presents a low oral risk to honeybees.

The HQcontact value is below the trigger value of 50, indicating that kasugamycin presents a low contact risk to

honeybees.

Pardosa spiders are known to be less sensitive than the recommended indicator species (Typhlodromus pyri and

Aphidius rhopalosiphi). Therefore this species is not the most suitable for the risk assessment. Therefore the

conclusion of the risk assessment has been based on predatory mites only.

The in-field HQ value for kasugamycin is above the trigger value of 2 for Typhlodromus pyri, (the most relevant

species), indicating that this active substance is of high concern for non-target arthropods.

The off-field HQ value for kasugamycin is below the trigger value of 2, indicating that this active substance is of low

concern for non-target arthropods.

The study on Typhlodromus pyri shows that sub-lethal concentrations have a significant effect on reproduction. So

even if the HQ is below the trigger for off-field situations on the basis of mortality, the off-field population may still be

at risk due to the effects on reproduction.

No higher tier study is available to evaluate the potential of recovery and/or recolonisation.

Medium

Disposal Death or

adverse effects

to aquatic or

terrestrial

organisms

Highly

improbable

Minor Negligible The applicant indicates that all attempts should be made to utilise

the substance completely in accordance with its registered use. In

all cases of disposal, the substance must be disposed of in

accordance with the requirements of the Hazardous Substances

(Disposal) Regulations 2001.

Negligible

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Relationship of Māori to the Environment

6.9. The potential effects on the relationship of Māori to the environment have been assessed in

accordance with sections 6(d) and 8 of the HSNO Act. Under these sections all persons

exercising functions, powers, and duties under this Act shall take into account the relationship

of Māori and their culture and traditions with their ancestral lands, water, taonga and the

principles of the Treaty of Waitangi (te Tiriti o Waitangi).

6.10. As outlined in Table 1, the hazards of Kasumin have the potential to inhibit Māori in fulfilling

their role of kaitiaki particularly in regards to the negative affect to the general health and well-

being of individuals and the community.

6.11. Based on the assessment of human health and environmental risks of Kasumin outlined in

Tables 2 and 3 above, and when considering information provided relating to the proposed use

pattern, the staff consider that the risks to the relationship of Māori to the environment are likely

to be negligible.

6.12. In addition, Kasumin may enhance the ability of Māori to fulfil their role as kaitiaki by providing

benefits such as reducing the incidence of Psa and replacing more harmful pest control

substances.

6.13. Based on the assessment for Kasumin outlined in Tables 2 and 3 above, and when considering

information provided relating to the proposed use pattern, the staff consider that the risks to the

relationship of Māori to the environment are likely to be negligible.

6.14. Given this assessment, there is no evidence to suggest that the use of Kasumin in accordance

with controls, will breach the principles of the Treaty of Waitangi.

Assessment of risks to society and the community and the market

economy

6.15. There are not expected to be any significant adverse impacts on the social environment with

the controlled use of Kasumin, apart from the health effects and environmental effects already

discussed. Consequently, the staff suggest that this aspect of potential risk need not be

considered further.

New Zealand’s international obligations

6.16. The staff did not identify international obligations that affect the approval of Kasumin.

Overall assessment of risks

6.17. With controls in place (as detailed in Section 7), the staff consider that the risks to human

health and the environment associated with Kasumin are mitigated. The overall level of risk is

therefore negligible.

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September 2013

Identification of benefits

6.18. The applicant has stated that the use of Kasumin may provide the following benefits:

the use of a new antibiotic ingredient that does not have additional uses as a human or

veterinary medicine, thus avoiding any issues with antibiotic resistance;

reducing the economic damage caused by Psa to the kiwifruit industry; and

replacing more harmful current control methods (e.g. streptomycin-based products).

The effects of the substances being unavailable

6.19. If Kasumin were unavailable, the benefits associated with the availability of the substance

would not be realised. In particular, the kiwifruit industry would not gain access to a new tool

which they consider very important for the control of Psa.

Overall assessment of benefits

6.20. The staff are satisfied that the availability of Kasumin will provide significant (i.e. non-negligible)

beneficial effects for some businesses and users of the substance, given the adverse effects

that Psa presents to the kiwifruit industry.

7. Controls

7.1. Based on the hazard classification determined for Kasumin, a set of associated default controls

specified by regulations under the Act has been identified by the staff as being applicable. The

default controls form the basis of the controls set out in Appendix C. Based on their risk

assessment, the staff consider that the additions, variations and deletions set out below are

applicable to Kasumin.

The setting of exposure limits

7.2. Tolerable Exposure Limits (TELs) can be set to control hazardous substances entering the

environment in quantities sufficient to present a risk to people. No TELs have been set for any

component of Kasumin at this time as it is considered that the risk to bystanders is considered

negligible. The EPA is however, required to set ADE and PDE values for new active ingredients

that may become present in food, to allow the setting of Maximum Residue Levels (MRLs) by

MPI. The staff propose setting the following ADE and PDE values:

Acceptable Daily Exposure (ADE) = 0.113 mg/kg bw/d

Potential Daily Exposure (PDEfood) = 0.08 mg/kg bw/d

Potential Daily Exposure (PDEdrinking water) = 0.023 mg/kg bw/d

Potential Daily Exposure (PDEother) = 0.11 mg/kg bw/d

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September 2013

7.3. The EPA typically adopts Workplace Exposure Standard (WES) values listed in the Ministry of

Business, Innovation and Employment’s WES Document5 to control exposure in places of work.

MBIE has set a WES value for Component D, but due to the low concentrations at which it is

present in the formulation, this has not been applied to Kasumin.

Variation and deletion of controls

7.4. The staff note that the environmental exposure modelling indicates there may be a risk where

the substance is used outside the specific parameters of the risk assessment. It is therefore

considered appropriate to set maximum application rates under section 77A:

Kasumin may be applied at a maximum application rate of 100 g kasugamycin/ha per

application; and up to four applications of Kasumin per year, with a minimum interval of 10 days

between applications.

Additional controls

7.5. The staff note that the environmental risk assessment indicates that restrictions on use are

necessary to ensure that the risks Kasumin presents to the aquatic environment are negligible,

and to mitigate risks to terrestrial invertebrates. Accordingly, it is considered that the application

of controls addressing these risks will be more effective than the default controls in terms of

their effects on the management, use and risks of the substance. Consequently, the following

additional controls are applied to Kasumin to restrict the level of risk to the environment:

Kasumin must not be applied onto, over or into water; and

Kasumin must only be applied via ground-based methods.

7.6. The staff consider that users of Kasumin need to be aware of the active ingredient contained in

the substance in order to be aware of the substance’s associated risks. Additionally, due to the

high concerns around the risks posed to non-target arthropods, the staff consider that the label

should include a statement identifying the potential risk to beneficial organisms. Accordingly,

the staff proposed that the following statement must appear on the label:

Kasumin contains the antibiotic, kasugamycin, and may be harmful to beneficial organisms

such as predatory mites, earthworms, and bees and other pollinator species.

7.7. The staff note that the specified controls do not address the risks associated with storage or

use of the substances within stationary container systems (e.g. tanks). These risks include the

failure of primary containment resulting in a large spill of the substance into the environment. In

addition, the default controls do not allow for dispensation where it is unnecessary for any

pipework associated with the stationary container systems to have secondary containment.

Accordingly, the application of controls addressing these risks are considered more effective

5 http://www.business.govt.nz/healthandsafetygroup/information-guidance/all-guidance-items/workplace-exposure-standards-and-biological-exposure-indices/workplace-exposure-standards-and-biological-indices-2013.pdf

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September 2013

than the specified default controls in terms of their effect on the management, use and risks of

the substance.

7.8. The staff note that the bystander risk assessment has identified potential risks to workers

entering an area after application of Kasumin. In order to mitigate this risk, a Restricted Entry

Interval (REI) is proposed.

7.9. An REI is the period of time which must elapse after application of a substance before entry

into the treated area is permitted without use of PPE or RPE. Entry into an application area

before the REI has elapsed is only permitted if the same level of PPE and RPE required for

application of the substance is worn. Where a substance is used in an indoor setting (such as a

greenhouse), the atmosphere may present an exposure risk for a period of time after

application has been completed. In such instances, the REI commences when the ventilation of

the building or structure commences, which may be by mechanical or passive means. The

person in charge of the place where a substance is applied is responsible for ensuring that no-

one enters the application area until the end of the REI.

7.10. In order to protect bystanders and/or sensitive areas from exposure arising from off-target

deposition of the substance, application of the substance is permitted provided that measures

are implemented by the applicator to ensure that off-target deposition (i.e. spray drift) is

sufficiently minimised to ensure that adverse effects beyond the property boundary do not

occur. The control is intended to provide applicators with the flexibility to adopt whatever drift-

mitigation measures are appropriate to their situation, and could include use of particular

application technologies or techniques. Or it could involve the use of shelter belts, and include

(but are not limited to) the New Zealand Standard NZS8409:2004 Management of

Agrichemicals details various spray drift reduction measures that can be implemented.

7.11. The use and details of the spray drift mitigation measures implemented should be included in

the records of applications. Accordingly, the requirements of control T3 that specify the details

included in the records of use (regulation 6 of Hazardous Substances (Classes 6, 8, and 9

Controls) Regulations 2001) is amended to include details of the spray drift mitigation

measures implemented.

7.12. The staff note that the environmental risk assessment highlighted several areas of potential

risk. The staff consider that an appropriate way to mitigate some of this risk is by ensuring all

persons applying the substance are qualified to do so. An approved handler requirement is

proposed to mitigate this risk.

7.13. The revised controls are shown in Appendix C.

7.14. The staff consider that the application of these controls will be more effective than the specified

(default) controls in terms of their effect on the management, use and risks of Kasumin (section

77A(4)(a)).

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September 2013

Environmental user charges

7.15. The staff consider that use of controls on Kasumin are an effective means of managing risks

associated with this substance. Therefore, it is not considered necessary to apply

environmental user charges to this substance as an alternative or additional means of

achieving effective risk management. Accordingly, no report has been made to the Minister for

the Environment.

Review of controls for cost-effectiveness

7.16. The staff consider that the proposed controls are the most cost-effective means of managing

the identified potential risks and costs associated with this application.

8. Overall evaluation and recommendation

8.1. The staff’s risk assessment indicates that use of Kasumin presents negligible risk to human

health, and non-negligible risks to the environment with controls in place.

8.2. The staff consider that, with the proposed controls in place (Appendix C), the overall level of

benefit provided by the availability of this substance is sufficiently great to outweigh the non-

negligible environmental risk, and recommend that the application be approved in accordance

with clause 27, and be subject to the controls proposed.

20


September 2013

Appendix A: Staff classification of Kasumin

The applicant submitted formulation test data for some endpoints of Kasumin. For endpoints where no

formulation data was provided, the staff have classified Kasumin using mixture rules as described in

the User Guide to Thresholds and Classifications6.

Table 1: Classifications of Kasumin

Hazard Endpoint Applicant Staff

Acute toxicity - 6.1E (dermal, inhalation)

Eye irritancy 6.4A No

Reproductive/developmental

toxicity -

6.8B

Target organ systemic toxicity - 6.9B

Aquatic toxicity 9.1C No

Data quality – overall evaluation

The data used by staff to classify Kasumin are the classifications which have been officially gazetted

during the transfer process and are publicly available through the HSNO Chemical Classification

Information Database (CCID)7. Where additional data have been considered for the risk assessment

of Kasumin, the EPA has adopted the Klimisch et al (1997)8 data reliability scoring system for

evaluating data used in the hazard classification and risk assessment of chemicals.

The staff have assigned Klimisch data reliability scores to submitted studies.

The staff acknowledge that there are frequently data gaps in the hazard classification for chemicals

which have been in use internationally for a long time. International programmes such as the OECD

High Production Volume programme9, REACH10, and European Regulation 1107/2009/EC11 are

progressively working towards filling these data gaps. As new information becomes available, staff will

update the Hazardous Substances and New Organisms (HSNO) classifications for those substances.

6

http://www.epa.govt.nz/Publications/ER-UG-03-2.pdf 7 http://www.epa.govt.nz/search-databases/Pages/HSNO-CCID.aspx 8 Klimisch, H-J., Andrear, M., & U. Tillmann, 1997. A systematic approach for evaluating the quality of experimental

toxicological and ecotoxicological data. Reg. Toxicol. Pharmacol. 25, 1–5 (1997) 9 http://www.icca-chem.org/Home/ICCA-initiatives/High-production-volume-chemicals-initiative-HPV/

10 http://ec.europa.eu/environment/chemicals/reach/reach_intro.htm

11 http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2009:309:0001:0050:EN:PDF

http://www.epa.govt.nz/Publications/ER-UG-03-2.pdf

http://www.icca-chem.org/Home/ICCA-initiatives/High-production-volume-chemicals-initiative-HPV/

http://ec.europa.eu/environment/chemicals/reach/reach_intro.htm

http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2009:309:0001:0050:EN:PDF

21


September 2013

Physico-chemical properties of the active ingredient

Table 2: Physico-chemical properties of kasugamycin

Property Test result Klimisch Score

(1-4) Reference

Colour Orange to Pale

brown power. 2

Health Canada, Proposed

registration decision PRD2012-

30 Kasugamycin. 27/11/2012

Physical state Solid

Odour Odourless 2




Oxidizing properties No data

pH 4.35 at 24.5ºC 2 US EPA Pesticide Fact Sheet:

Kasugamycin. September 2005.

Explosive properties

Henry’s Law

constant

2.44 × 10-13

atm·m3·mole

-1

2




Melting range 202–230

oC

(decomposition) 2




Vapour pressure < 0.013 mPa 2




Relative Density 0.40–0.46 g/mL 2




Viscosity

Surface Tension

Water Solubility

pH 5: 20.7 g/100

mL

pH 7: 22.8 g/100

mL

pH 9: 43.8 g/100

mL

2




Solvent Solubility

(25°C)

Methanol 0.744

g/100 mL

Hexane < 1 × 10-

5 g/100 mL

Acetonitrile < 1 ×

2




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September 2013

10-5

g/100 mL

Methylene

chloride < 1 × 10-

5 g/100 mL

Log Kow < -1.96 at pH5 2




Flammability

Auto flammability

Mammalian toxicology - Robust study summaries for the active

ingredient and metabolite

Acute toxicity

Acute Oral Toxicity [6.1 (oral)]

Type of study: Acute oral toxicity in the rat.

Flag: Key study

Test substance: Kasugamycin technical

Endpoint: LD50, signs of toxicity

Value: > 5000 mg/kg bw

Reference: Glaza S.M.; 1992. Acute oral toxicity study of kasugamycin hydrochloride technical in rats

(EPA Guidelines). Laboratory Project Identification HWI 20504630. Hazleton Wisconsin Inc, 3301

Kinsman Boulevard, Madison, Wisconsin 53704, USA.

Klimisch Score: 1

Amendments/Deviations: None

GLP: Yes

Test Guidelines: US EPA Guideline 81-1

Species: Rats

Strain: Albino, Crl CD BR

No/Sex/Group: 5

Dose Levels: 5000 mg/kg bw

Exposure Type: Oral by gavage

Study summary: The animals appeared normal throughout the study and gained the anticipated body

weight. There were no clinical signs of toxicity during the study or visible lesions at gross necropsy.

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September 2013

Conclusion: The substance does not trigger classification for acute toxicity via the oral route.

Acute Dermal Toxicity [6.1 (dermal)]

Type of study: Acute dermal toxicity in the rabbit

Flag: Key study

Test Substance: Kasugamycin technical



Reference: Glaza S.M.; 1992 Acute dermal toxicity study of kasugamycin hydrochloride technical in

rabbitts (EPA Guidelines). Laboratory Project Identification HWI 20504631. Hazleton Wisconsin Inc,

3301 Kinsman Boulevard, Madison, Wisconsin 53704, USA.

Klimisch Score: 1

Amendments/Deviations: EPA staff note the report contains errors without the correction being

provided (this is on p16).

GLP: Yes

Test Guidelines: US EPA Guidelines 81-2

Species: Rabbits

Strain: Hra: (New Zealand White) SPF

No/Sex/Group: 5


Exposure Type: Semi occlusive patch test.

Study Summary: The animals appeared clinically normal during the study with the exception of a

slight erythema reaction in one female on Day 1. Study authors state: “There was no meaningful

effect on body weight gain.” [EPA staff note this statement cannot be verified as two errors in

recording of body weight are indicated on p16 and the corrected values are not displayed.

Nevertheless, all other animals gained weight and in view of the lack of clinical signs this error is not

considered to invalidate the report. The study report indicates these are recording errors rather than

an indication of a substance-effect on body weight.]

Conclusion: The substance does not trigger classification for acute dermal toxicity.

Acute Inhalation Toxicity [6.1 (inhalation)]

Type of study: Acute inhalation in the rat

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September 2013

Flag: Key study

Test Substance: Kasugamycin technical (Lot #KP-459) [Reported in the study to consist of 84.8%

kasugamycin hydrochloride hydrate and 15.2% other ingredients].

Endpoint: LC50, signs of toxicity

Value: >2.07 mg/L

Reference: Durando, J (2009) Kasugamycin Technical: Acute inhalation toxicity study in rats – limit

test. Laboratory Study No. 25917. Eurofins Product Safety Laboratories, 2394 US Highway 130,

Dayton, NJ 08810, USA

Klimisch Score: 1


GLP: Yes

Test Guidelines: US EPA Health Effects Test Guidelines OPPTS 879.1300 (1998)

Species: Rats

Strain: Sprague Dawley, albino

No/Sex/Group: 5

Dose Levels: 2.07 mg/L

Exposure Type:

Whole animal/nose exposure: Whole body exposure

Mass Median Aerodynamic Diameter (MMAD) and Geometric Standard Deviation: 3.8 µm ± 2.05

Study Summary: All animals survived the exposure and gained weight over the 14 day period.

In chamber observations included hunched posture and hypoactivity. Three hours into the exposure,

in-chamber animal observations were limited due to the accumulation of test substance on the walls

of the exposure chamber. All animals recovered from the above clinical signs upon removal from the

exposure chamber and appeared active and healthy over the 14-day observation period. No gross

abnormalities were noted for any of the animals when necropsied at the conclusion of the 14-day

observation period.

Conclusion: The substance does not trigger classification for acute inhalation toxicity.

Comment on classification: The achieved concentration is lower than the cut off for classification

category 6.1D (5 mg/L), however given that the requirement to test at a MMAD between 1 and 4 µm

corresponds to a maximum concentration of about 2 mg/L, and the lack of signs of toxicity, the

substance should not be classified. The method (whole body exposure) raises questions, particularly

in view of the comment about the coating of the inside of the animal exposure chamber impeding

cage side observation, but this is likely to give rise to additional oral or dermal exposure of the

25


September 2013

animals, so would raise uncertainty if an inhalation classification were proposed rather than for the

current null outcome.

Skin Irritation [6.3/8.2]

Type of Study: Skin irritation/corrosion in rabbits

Flag: Key study



Endpoint: Mean Draize Score

Value: 1.67 for both erythema and oedema

Reference: Durando J.; 2009 Kasugamycin Technical: Primary Skin Irritation Study in Rabbits.

Laboratory Study No. 25919. Eurofins Product Safety Laboratories. 2394 US Highway 130. Dayton.

NJ 08810. USA. [For Arysta Lifescience North America LLC]

Klimisch Score: 1

Amendments/Deviations: Due to an oversight the animals were scored and observed through Day 10.

This had no impact on the outcome of the study.

GLP: Yes


Species: Rabbit

Strain: New Zealand White

Mean Draize Score:

Erythema: 15/9 = 1.67

Oedema: 15/9 = 1.67

No/Sex/Group: 3 F

Dose: 0.5 ml

Study Summary:

For the first 48 hours after patch removal all three sites exhibited well defined erythema and slight

oedema. The incidence and severity reduced with time and erythema and oedema had cleared by

day 7. The Mean Draize scores were:

Erythema: 15/9 = 1.67

Oedema: 15/9 = 1.67

One animal had desquamation at the dose site seen on Day 7 only (which had cleared by Day 10).

26


September 2013

The Mean Draize Scores are greater than the 1.5 value triggering for these endpoints.

Conclusion: Kasugamycin in classified as irritating to skin (6.3B).

Eye Irritation [6.4/8.3]

Type of Study: Eye irritation/corrosion in rabbits.

Flag: Key study




Value: 0.44 for conjunctival redness. Corneal opacity 0.11.

Reference: Durando J.; 2009. Kasugamycin Technical: Primary Eye Irritation Study in Rabbits.

Laboratory Study No. 25918. Eurofins Product Safety Laboratories. 2394 US Highway 130. Dayton.

NJ 08810. USA. [For Arysta Lifescience North America LLC]

Klimisch Score: 1


GLP: Yes


Species: Rabbits


No/ Sex/Group: 3 F

Dose: 0.1ml

Study Summary:

The score for conjunctival redness was: 5/9 = 0.56

The score for conjunctival chemosis was: 0/9 = 0.0

Corneal opacity was graded one in one animal at 24 hours (giving a score of 0.11), but subsequently

cleared at 48 hours.

The irises were unaffected by treatment at all time points.

All animals were free from eye irritation within 72 hours.

Conclusion: Under the HSNO classification system the substance does not trigger classification as

an eye irritant.

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September 2013

Contact Sensitization [6.5]

Type of study: Contact sensitisation in the guinea pig (Buehler method)

Flag: Key study



Endpoint: Sensitisation

Value: Negative

Reference: Durando J.; 2009 Kasugamycin Technical: Dermal Sensitisation Study in Guinea Pigs

(Buehler Method). Laboratory Study No. 25920. Eurofins Product Safety Laboratories. 2394 US

Highway 130. Dayton. NJ 08810. USA. [For Arysta Lifescience North America LLC]

Klimisch Score: 1


GLP: Yes


Species: Guinea pigs

Strain: Hartley, albino

No/Sex/Group: 20 in the challenge test (males and females), 10 in the naïve controls (males and

females), 4 in the preliminary irritation challenge (males and females)

Type of Exposure: Semi-occluded patch. The induction phase used a 65% w/w mixture of the

substance in distilled water. The challenge phase used a 33% w/w mixture of the substance in

distilled water.

Study Summary: No significant erythema was seen during induction or challenge.

Conclusion: The substance does not trigger classification for contact sensitisation.

General conclusion about acute toxicity

The substance does not trigger for classification as an acute toxicant via the oral, dermal or inhalation

routes. It does not require classification for eye irritation or contact sensitisation, but does trigger

classification as a skin irritant (6.3B).

Genotoxicity [6.6]

In Vitro Study

Type of Study: Mutagenicity in microbial systems

Flag: Supporting study

Test Substance: Kasugamycin hydrochloride (80% purity)

28


September 2013

Endpoint: Mutagenicity

Value: Negative [EPA Staff tentative conclusion]

Reference: Shirasu, Y, et al.; 1976. Mutagenicity Testing on Kasugamycin HCl in Microbial Systems.

Institute of Environmental Toxicology. Toxicology Division. [No address given]

Klimisch Score: 2 EPA Staff note this study predated test guidelines and GLP.

Amendments/Deviations: Not applicable

GLP: EPA Staff note this study predated test guidelines and GLP.

Test Guidelines: EPA Staff note this study predated test guidelines and GLP.

Cell Type: Bacillus subtilis, Escherichia coli WP2 and five strains of Salmonella typhimurium TA

(TA1535, TA1537, TA1538, TA98 and TA100) with and without metabolic activation, and in host

mediated assay with S typhimurium in mice.

Dose Rate: In the recombination assay the dose range was 20 – 2000 µg/plate. In the in vitro tests

the range of concentration tested was 5 – 200 µg/plate. In the in vivo host study the dose range was

up to 200 and 5000 mg/kg bw. [EPA staff note that the range tested in the in vitro test is lower than

that subsequently approved in guidelines in the absence of cytotoxicity.]

Response: Negative

Conclusion: No evidence of mutagenicity, but due to the age of the study (predating test guidelines)

this conclusion is tentative.

Type of Study: Bacterial cell mutation assay

Flag: Key study

Test Substance: Kasugamycin technical (71.5% active ingredient)


Value: Negative

Reference: Wagner VO and VanDyke MR (2009) Kasugamycin technical: Bacterial reverse mutation

assay. BioReliance study number AC29CA.503.BTL. BioReliance, 9630 Medical Center Drive,

Rockville MD 20850 USA.

Klimisch Score: 1

Amendments/Deviations: Due to a lack of dose formulation analyses the interpretation of the study

data was based on the nominal dose levels rather than actual test article concentrations as confirmed

by analytical determinations. However, the authors considered that the toxicity observed in the assay

indicated that the substance was tested up to the required regulatory level.

GLP: Yes

29


September 2013

Test Guidelines: US EPA OPPTS 870.5100 (1998) OECD Test Guideline 471 (1998)

Cell Type: Salmonella typhimurium strains TA98, TA100, TA1535, TA1537; Escherichia coli strain

WP2 uvrA

Dose Rate: TA 1537: 15, 50, 150, 500, 1500 and 5000 µg kasugamycin per plate; WP2 uvrA: 1.5, 5.0,

15, 50, 150, 500 and 1500 µg kasugamycin per plate; TA98, TA100 and TA1535: 5.0, 15, 50, 150,

500 and 1500 kasugamycin per plate (with and without metabolic activation)

Response: In both the initial toxicity-mutation assay and the confirmatory mutagenicity assay no

positive mutagenic responses were seen in any strain in the presence or absence of S9 metabolic

activation.

Conclusion: No evidence of mutagenicity

Type of Study: In vitro cytogenicity assay in mammalian cells


Test Substance: Kasugaymcin technical (purity 67.1% Lot no. KP-570)




Value: Negative

Reference: Ivett, J.; 1985. Mutagenicity Evaluation of Kasugamycin Tech in an in vitro Cytogenetic

Assay Measuring Chromosome Aberration Frequencies in Chinese Hamsters Ovary (CHO) Cells. LBI

Project No 20990. Litton Bionetics Inc. 5516 Nicholson Lane, Kensington, Maryland 20895, USA.

Klimisch Score: 2

Amendments/Deviations: Not applicable

GLP: Yes

Test Guidelines: No evidence of compliance with test guidelines (but appears to have been conducted

in line with OECD TG 473).

Cell Type: Chinese hamster ovary cells.

Dose Rate: 1 – 5 mg/ml in both assays (with and without metabolic activation).

Response: There was no toxicity at the top dose with or without metabolic activation. There was no

increase in the percentage of cells with chromosomal aberrations either with or without metabolic

activation.

Conclusion: No evidence of induction of chromosomal aberrations, but due to this being a non

guideline study conclusion is tentative.

30


September 2013

Type of Study: Chromosomal aberration study in vitro

Flag: Key study

Test Substance: Kasugamycin technical (71.5% active ingredient)

Endpoint: Chromosomal aberrations

Value: Negative

Reference: Gudi R and Jois M (2009) Kasugamycin technical: in vitro mammalian chromosome

aberration test. BioReliance study number AC29CA.331.BTL. BioReliance, 9630 Medical Center

Drive, Rockville MD 20850 USA.

Klimisch Score: 1

Amendments/Deviations: Due to a lack of dose formulation and stability analyses the interpretation of

the study data was based on targeted concentrations as verified by formulation verification.

GLP: Yes

Test Guidelines: US EPA OPPTS 870.5375 (1998) OECD Test Guideline 473 (1998)

Cell Type: Chinese hamster ovary (CHO)

Dose Rate: Preliminary toxicity assay: up to 4340 µg/L (10 mM); Main assay: 542.5-4340 µg/L

Response: The percentage of cells with structural or numerical aberrations in the test substance

treated groups was no significantly increased relative to the solvent control at any dose level, in the

presence and absence of S9 activation.

Conclusion: No evidence of induction of chromosome aberrations.

Type of Study: Unscheduled DNA synthesis in human cells.


Test Substance: Kasugaymcin technical (purity 67.1% Lot no. KP-570)



Endpoint: Unscheduled DNA synthesis.

Value: Negative is the authors’ conclusion, based on the third assay using s9 being negative, but the

second assay with s9 the result was positive, while the first assay was negative at a lower dose

regime.

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September 2013

Reference: Seeburg, A. H. and Forster, R.; 1985. Unscheduled DNA Synthesis in Human Cells: Cell

Line Hela S3: Test substance: Kasugamycin Tech. LSR-RTC report No: 161001-M-01885. Life

Science Research, Roma Toxicology Centre, Via Tito Speri, 14, 00040 Pomezia (Roma), Italy.

Klimisch Score: 2

Amendments/Deviations: EPA stuff note that some samples were lost in the first two experiments,

while in the third, some cultures had to be discarded due to microbial contamination.

GLP: Yes

Test Guidelines: No evidence of compliance with test guidelines.

Cell Type: Human cells (HeLa S3

Dose Rate: 10.0, 5.0, 2.5, 1.25, 0.625, 0.313, 0.156 mg/ml (see details below for the three assays).

Response:

The preliminary cytotoxicity test used 10 mg/ml and 9 spaced lower doses. The test substance was

cytotoxic and caused signs of toxicity at the higher dose levels. The maximum dose recommended for

use in the UDS assay was 2.5 mg/ml.

1st study used: 2.5, 1.25, 0.625, 0.313 and 0.156 mg/ml. The first assay was negative with no signs of

cytotoxicity so the subsequent experiments used a different dose regime.

2nd

and subsequent: 10.0, 5.0, 2.5, 1.25, 0.625, and 0.313 mg/ml.

The 2nd

assay gave a dose-related increase in thymidine incorporation in the s9 series. In the

absence of S9, incorporation was greater than 50% of control values at one dose only (0.313 mg/ml).

A third experiment using only the s9 exposures was carried out and no marked increase in thymidine

incorporation was observed. The study authors had no explanation for the inconsistent results, but

concluded that the substance does no induce an increase in UDS either with or without s9 metabolic

activation.

EPA staff note that the same s9 mix was used for the 2nd

and 3rd

experiments the age of the

preparation might be an issue, but it did produce the appropriate response in the Benzo(a)pyrene

positive control (requiring s9 activation).

These are the results for comparison purposes:

Ratio of UDS relative to solvent all determination as the mean of three replicates.

Test 1 Test 2 Test 3

(w/o s9) (with s9) (w/o s9) (with s9) (with s9)

Solvent 1** 1 1 1 1.0

0.156 1.1 1.1 - - -

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September 2013

0.313 1.1 1.2 1.6 1.5 0.9

0.625 1.0 1.1 1.3 1.7** 0.9**

1.25 1.1 1.1 1.2 1.7** 1.0

2.5 1.1 1.3 1.0 2.0 0.9**

5.0 - - 1.2 1.7 0.8

10.0 - - 0.8 1.4 0.7

Positive

control*

18.1 1.7 17.4 2.3 2.0

* The positive controls are: 4 nitroquinoline-N-oxide (direct acting) and benzo(a) pyrene (for mutagens

needing s9 activation).

** One of three replicates lost/discarded.

Conclusion: The results are equivocal. No evidence of an increase in DNA synthesis was seen in the

final dosage regime in the third assay with s9, after the second assay gave a response. EPA staff

were not convinced by this study, but noted that the US EPA Fact sheet appears to accept this study

without reservation as did the Canadian PMRA.

In Vivo Study

Type of Study: Chromosomal abnormalities in mice (in vivo)

Flag: Key study

Test Substance: Kasugamycin technical (KP-570)


Value: Negative for clastogenicity

Reference: Bootman, J; et al.; 1985. Kasugamycin: Assessment of clastogenic action on bone

marrow erythrocytes in the micronucleus test. Laboratory Report No 85/HKC001/207. Life Science

Research, Eye, Suffolk, IP23 7PS, England.

Klimisch Score: 1


GLP: Yes

Test Guidelines: No guidelines cited (but appears to be consistent with OECD TG 474 apart from the

scoring of a smaller number of ethryocytes than is currently recommended).

Species: Mice

33


September 2013

Strain: CD-1

No/Sex/Group: 15 for controls and top dose, 5 for 200 and 1000 mg/kg bw (and positive control

chlorambucil), 4 in preliminary study

Dose Rate: 0, 200, 1000, or 5000 mg/kg bw, by gavage

Response: In the preliminary study one male animal exhibited hunched posture, lethargy and

piloerection at 5000 mg/kg bw two days following treatment. Frequencies of micronucleated

polychromatic erythrocytes in animal exposure for 24, 42 or 72 hours were similar to those of

concurrent vehicle controls in both sexes. The positive control gave the anticipated response.

Conclusion: The substance does not trigger classification for mutagenicity.

General conclusion about genotoxicity

Negative results were found in all but one of the genotoxicity studies available. There is an equivocal

finding for the in vitro unscheduled DNA synthesis test, but that in vitro finding would need

confirmation from an in vivo test to justify classification. While a number of studies are old and pre

date test guidelines, two more recent in vitro studies are also available which report negative results.

Based on the negative findings in all the other in vitro and in vivo studies, and taking into account the

lack of developmental toxicity or any evidence of carcinogenicity in bioassays in rats and mice, EPA

staff consider that the data are sufficient to conclude that the substance should not be classified for

mutagenicity.

Carcinogenicity [6.7]

Type of Study: Chronic toxicity and carcinogenicity in the rat.

Flag: Key study

Test Substance: Kasugamycin technical (lot number KP-570, purity 67.1%)

Endpoints:

a. Non-neoplastic effects

LOAEL: 3000 ppm (equivalent to 116 and 140 mg/kg bw/day in males/females respectively) based

on organ weight and associated histopathological changes in the cecum and proximal tubules of

the kidney. [Testicular softening and atrophy also identified by US EPA and PMRA.]

NOAEL: 300 ppm (equivalent to 11.31 and 13.42 mg/kg bw/day in males/females respectively)

b. Neoplastic effects

LOAEL: >3000 ppm (no increase in tumours associated with the substance reported).

NOAEL: 3000 ppm

Tumours: N/A

Malignant/ Benign: N/A

Background Incidence: N/A

Time of Onset: N/A

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September 2013

Survival: N/A

Dose/ Response: None

Reference: Kitazawa, T, 1987. Kasugamycin: 24 month oral chronic toxicity and oncogenicity study in

rats. The Institute of Environmental Toxicology, Mitsukaido Laboratories, Uchimariya 4321,

Mitsukaidao, Ibaraki 302-02, Japan. [EPA Staff note there is not test facility report number.]

Klimisch Score: 2

Amendments/Deviations: A 46 page summary has been provided by the applicant, rather than the full

study report.

GLP: Yes

Test Guidelines: No test guidelines cited.

Species: Rat

Strain: Wistar

No/Sex/Group: 70 (10 of which were examined at interim kill at 26 and 52 weeks – a total of 20

animals).

Dose Levels: 0, 30, 300, 3000ppm equivalent to 1.146, 11.31, and 116.0 mg/kg bw/day in males and

1.371, 13.42, and 139.8 mg/kg bw/day in females.

Exposure Type: Oral via diet

Study Summary:

A preliminary 4 week range finding study used dietary levels of 0, 300, 1000, 6000 and 10000 ppm. In

this test the males and females at dose of 6000 ppm and above showed: decreases in body weight

gain, food consumption, and food efficiency; an increase of water consumption, decreases in total

protein, albumin and globulin. Increase in kidney salivary gland and cecum weights, and

histophathological abnormalities in the kidney and salary glands. At 3000 ppm, males showed

increased water consumption, atrophy and degeneration of the proximal tubular cells in the kidneys.

In the main study the following finding were reported.

At 3000 ppm retarded growth was found in the female animals, during the first year of the study.

Blood clinical testing showed decreased alkaline phosphatase in both sexes, reduced total protein

and globulin in males and in total cholesterol in females. The organ weight analysis revealed

increased (absolute) [EPA staff assumption] organ weight of cecum and increased relative kidney

weight in both sexes. The increased cecum weight was considered to be due to the effect of

treatment with a large amount of antibiotics (Kasugamycin) on intestinal flora. In addition females

exhibited a decreased absolute and relative weight of liver and an increased relative weight of salivary

glands. Histopathology revealed increased brown deposition in the proximal tubular cells, identified as

lipfuscin, in the kidney and foam cell aggregation in the lung in males and females. The authors noted

that this lesion may be associated with disorders of lipid metabolism and treatment with some

35


September 2013

chemicals causing hypocholestrinemia, and that serum cholesterol levels were decreased in females

treated with 3000 ppm at weeks 52, 78 and 104. Increased incidences of rhinitis in males and

hepatocellular atrophy in females were observed.

In 300 and 30 ppm, there were no toxicological findings attributed to the test substance.

There was no evidence of any carcinogenicity for the substance in this study.

EPA staff note that the US EPA and PMRA summaries identify testicular softening and atrophy in this

study but this is not documented in the summary available. Nevertheless this finding has been added

to the adverse effects table as it is consistent with the other data.

Conclusion: The substance does not trigger classification for carcinogenicity.

Type of study: 78 week carcinogenicity study in mice

Flag: Key study

Test Substance: Kasugamycin (Batch number KP-821)

Endpoints:

a) Non-neoplastic effects

LOAEL: No toxicologically significant findings identified. [1500 ppm equivalent to 186.3 mg/kg

bw/day was initially identified as the LOAEL based on a reduction in the incidence of enlarged

spleen (in males only), but EPA staff noted that the US EPA data summary identifies this as the

NOAEL in both males and females, EPA staff consider this effect is not really of toxicological

significance, so this dose level should be considered the toxicological NOAEL.

NOAEL: 1500ppm, equivalent to 186.3 mg/kg bw/day (males) and 215.2 mg/kg bw/day in females.

b) Neoplastic effects

LOAEL: > 1500 ppm equivalent to >186.3 mg/kg bw/day in males and >215.2 mg/kg bw/day in

females

NOAEL: 1,500 ppm equivalent to 186.3 mg/kg bw/day in males and 215.2 mg/kg bw/day in

females

Tumours:

Malignant/ Benign: No tumour response seen

Background Incidence: No tumour response seen

Time of Onset: No tumour response seen

Survival: No tumour response seen

Dose/ Response: No tumour response seen

36


September 2013

Reference: Holmes, P; 1992 Kasugamycin: Oncogenicity Study by Dietary Administration to CD-1

Mice for 78 weeks. LSR Report 91/HKC006/1010. Life Science Research Limited, Eye, Suffolk IP23

7PX, England.

Klimisch Score: 1


GLP: Yes

Test Guidelines: OECD No 451 (1981), US EPA (FIFRA), 1984.

Species: Mouse

Strain: CD-1

No/Sex/Group: 72 (A maximum of 20 animals of each sex were selected for interim examination at 52

weeks).

Dose Levels: 0, 50, 300, 1500 ppm (in diet) equivalent to average achieved doses over the 78 weeks

of 5.93, 34.94 and 186.3 mg/kg bw/day (in males) and 7.25, 42.49, and 215.2 mg/kg bw/day (in

females).

Exposure Type: Dietary administration

Study Summary: There were no signs of behaviour and appearance that were considered treatment-

related.

Some males display penis mutilation and/or skin encrustations that were considered to be associated

with fighting.

A large proportion of the male animals assigned for the interim examination in the 1500 ppm group

died or were killed prematurely, but this was not considered a consequence of treatment, being

associated with lesions resulting from fighting. As no difference in mortality was seen in the terminal

phase this finding was not considered treatment-related.

Food consumption of males receiving 1500 ppm was marginally higher than controls throughout the

treatment period. This finding was of small and of doubtful toxicological significance. Over the first 14

weeks of treatment, the food utilisation was marginally lower in the males receiving 1500 ppm than

controls

The overall body weight gains of the treatment males was slightly higher and those of the treated

females slightly lower than respective controls. In neither case was there a dose-response

relationship, so this was considered a chance finding.

Haematological examination revealed no treatment-related effects.

In the males given 1500 ppm, there was slightly lower absolute spleen weight and spleen weight

relative to body weight. These findings were considered to represent a decreased incidence of a

37


September 2013

common abnormality (apparently large spleens) and did not correlate with any significant

histopathological changes, and were therefore not considered to be of toxicological significance.

Macroscopic pathology did not reveal any findings related to treatment, apart from a lower incidence

of enlarged spleens in males at 1500 ppm.

Histopathology indicated a higher incidence of extramedullary haematopoiesis in males receiving

1500 ppm.

There were no findings of tumours that were increase in a manner indicating a treatment-related

effect.

Conclusion: The study shows no evidence that the substance caused an increase in tumours in

mice.

General conclusion about carcinogenicity

There is no evidence that the substance causes an increase in tumours, and therefore it does not

require classification.

Reproductive/Developmental Toxicity [6.8]

Developmental Toxicity

Type of study: Teratogenicity study in rodents

Flag: Key study

Test Substance: Kasugamycin

Endpoints:

a. Maternal toxicity

LOAEL: 1000 mg/kg bw/day (based on reduced body weight gain and distention of the large

intestine with stool in the caecum)

NOAEL: 200 mg/kg bw/day.

b. Developmental (foetal) effects

LOAEL: > 1000 mg/kg bw/day (no adverse effects seen)

NOAEL: 1000 mg/kg bw/day

Reference: Fujii, S; 1991 Teratogenicity study in rats with Kasugamycin (Study no, IET 89-0085). The

Institute of Environmental Toxicology, Kodaira-Shi, Tokyo 187, Japan.

Klimisch Score: 2


GLP: Yes

Test Guidelines: No reference made to recognized guidelines

38


September 2013

Species: Rat

Strain: Crj: SC (SD)

No/Group: 24 (females)

Dose: 0, 40, 200, 1000 mg/kg bw/day from Day 6 – 15 of gestation.

Exposure Type: Oral by gavage.

Study Summary:

At 1000 mg/kg bw/day seven of 24 rats showed loose stool sporadically during the treatment period.

No significant differences in the body weights or adjusted weights were seen at 40 or 200 mg/kg

bw/day, but at 1000 mg/kg bw from gestation Day 8 (third day of dosing) though Day 20 (the day of

cesarean section) weights and adjusted weights were lower than those in the control group. From Day

8 – through Day 20 the body weight gain of the 1000 mg/kg bw/day group was significantly lower than

the control group.

The food intake of the 40 mg/kg bw/day group was unaffected by treatment, but the intake was

reduced for the 200 and 1000 mg/kg bw/day groups for Day 6 – 9. The food intake returned to control

levels for the 200 mg/kg bw/day, from 9- 12 Days and thereafter. For the 1000 mg/kg bw/day group

the food intakes remained significantly lower than controls, however, in the post-dosing period (Days

15 – 20) the food intake in this group was significantly higher than in the control group.

In the maternal animals the only gross pathological finding was of distention with stool in the caecum

seen in 5/24 animals at 1000 mg/kg bw/day.

There were no significant differences found in the number of live fetuses, percentage incidences of

fetal resorption and deaths, fetal sex rations, fetal and placental weights in the 40 and 200 mg/kg

bw/day groups. In the 1000 mg/kg bw/day group, although placental weights were significantly lower

than those in the control group, no adverse effects relating to treatment were observed in other

parameters. [EPA staff note the average weights for both male and female pups in the top dose group

were reduced, but not to a statistically significant extent.]

In the external, visceral and skeletal examination of live fetuses, no malformation or variations

attributable to treatment were observed. There was a slight increase in the incidence (3.9%) of

fetuses with shortening and/or absence of the 13th rib at 1000 mg/kg bw/day, but this incidence is

within historical background (0 – 4.19%). The total number of fetuses with variations was slightly

increased in the 200 mg/kg bw/day group but in the absence of a dose response this was not

considered toxicologically significant.

Conclusion: There was no evidence of any fetotoxic or teratogenic effects in this study.

Reproductive toxicity

Type of Study: 2 generation reproductive toxicity in the rat

39


September 2013

Flag: Key study

Test Substance: Kasugamycin (lot number KP-913)

Endpoints:

a. Parental toxicity

LOAEL: 6000 ppm in males and females due to red swollen skin around the anal opening and

macroscopic/microscopic ulceration at the boundary of the rectum and anus (at necropsy).

NOAEL: 1000 ppm (69.99 and 84.86 mg/kg bw/day in males and females respectively).

b. Reproductive toxicity

LOAEL: 6000 ppm (421.8 mg/kg bw/day) in males relating to reduced fertility, >6000 ppm (>505.17

mg/kg bw/day) in females [no adverse reproductive effects]

NOAEL: 1000 ppm in males, 6000 ppm in females

NOAEL (pups): 6000 ppm. [There were no adverse effects seen in the pups at all doses tested.]

Reference: Henwood, S. M.; 1993. Two-generation reproduction study with Kasugamycin in rats.

Laboratory Project Identification HWI 6434–102. Hazleton, Wisconsin, 3301 Kinsman Boulevard,

Madison, Wisconsin 53704, USA.

Klimisch Score: 1


GLP: Yes

Test Guidelines: US EPA FIFRA Guideline 83- 4. OECD Guideline No 416.

Species: Rat

Strain: Crl: CD BR VAF/Plus

No/Sex/Group: 25

Dose: 0, 200, 1000, 6000 ppm in diet (in F0 animals 10 weeks before mating then through mating,

gestation and lactation. Selected animals (from F1) were then exposed to the same dose as their

parent during mating, gestation and lactation of the F2 generation. The target dose levels were

equivalent to measured intake (at pre-mating phase (Weeks 0 – 10) of:

F0 (males): 13.56, 69.99 and 421.80 mg/kg bw/day respectively

F0 (females): 16.31, 84.86 and 505.17 mg/kg bw/day respectively

F1 (males): 13.87, 70.68 and 428.74 mg/kg bw/day respectively

F1 (females): 16.08, 80.92 and 501.71 mg/kg bw/day respectively

Exposure Type: Oral in diet

40


September 2013

Study Summary:

The only clinical sign considered exposure related was red swollen skin around the anal opening

which was seen in adult animals of both sexes in the F0 and F1 generations at 6000 ppm. The finding

was seen in 7/25 of F0 males, all F0 females 21/25 F1 males and all F1 females. The onset was first

seen at week 10 in the F0 adults. The findings were seen in Week 5 for the F1 generation. [EPA staff

note the authors report some exposure of the F1 animals in utero and via lactation may occur,

assumed to be a proposed explanation for the earlier on-set in these animals.]

There were four unscheduled deaths of adult animals, two F0 males at 1000 ppm, one F1 males at

1000 ppm and F1 male at 6000 ppm.

Mean body weight and cumulative body weight changes were lower for F0 males at 1000 and 6000

pm. Similar changes were not seen in the F1 males, apart from at 6000 ppm in the final F2b phase.

Mating and fertility indices were unaffected by the test material for the F0 generation. In the F1

generation dosed at 6000 ppm, reduced fertility was found. There were a significantly fewer pregnant

F1 females in that group for both both the F2a and F2b matings. None of the F1 males that failed to

sire a litter in F2a mating sired a litter in the F2b mating. [Note that the study specifically selected the

failed sires to examine their fertility again. It was also designed to select proven fertile females with

the failed males, demonstrating that the effect on fertility related to an effect on the males.] In addition,

of the 16 F1 males that sired litter during the F2a mating, only nine sired litters in the F2b mating. This

demonstrated a substance related reduction in fertility over time and across matings. The

histopathological investigation indicated confirmation of the adverse effect on seminiferous tubules in

the 6000 ppm males only. The description in the microscopic observation reads:

“Atrophy/degeneration, bilateral: characterised by complete low of germinal epithelium, atrophy of the

seminiferous tubules and persistence of sertoli cells. Variable amounts of inter-tubular granuloma

oedema were also frequently present.” This description applies to 15/24 males at 6000 ppm.

The male fertility index for the F1 males at F2a was 100 x (16/25) = 64%

The male fertility index for the F1 males at F2b was 100 x (9/25) = 36%

There values being very low in comparison to controls, 200 and 1000 ppm groups which ranged

between 88- 96%.

The mean co-habiting days for the 1000 ppm group for the F2a mating were significantly shorter than

controls, but this was not considered substance related by authors, as no similar finding occurred at

6000 ppm. However, mean cohabitation days of the 6000 ppm for the F2b mating were significantly

lower than those of control which indicates a possible effect on libido.

Examination of uteri from the F1 females did not indicate any adverse effect, although due to two

reproductive cycles meaningful interpretation was difficult. Data were inconclusive.

There were no test substance-related effects noted for gestation, viability, or weaning indices for F1,

F2a or F2b litters. There were no differences in the observed number of pups per litter or the sex

ratio.

41


September 2013

The covariate adjusted mean body weights of the F1 females pups from the 6000 ppm group on

Lactation Day 21, were significantly less than those of the control group. The authors were uncertain

whether this finding is substance-related as a significant difference was not seen for the male pups of

the generation (F2a) or in pups of either sex in the F2b generation.

Macroscopic and microscopic changes seen at 6000 ppm included ulceration associated with chronic-

active inflammation in the rectum and rectal anal junction in F0 and F1 adults and bilateral

degeneration and loss of germinal epithelium and the seminiferous tubules of the testes with tubular

atrophy and some inter tubular oedema in F1 males.

The NOAEL is 1000 ppm for the effects on reproductive performance (based on reduced fertility in

males).

The NOAEL for pup viability and growth was 6000 ppm.

EPA Staff note there appears to be bias towards lower than intended concentration (in preparation of

the diets), since at 200 and 6000 ppm respectively the homogeneity tested results ranged from 87.5 –

96.5% and 91.5 - 95.8% respectively of the target concentrations. This should be taken into account if

the study is used for derivation of any critical endpoints.

In deciding on the classification, EPA staff note that the adverse effects on male fertility were only

seen at the highest dose tested which is the LOAEL for other toxicological findings, nevertheless, in

view of the severity of the effect, the classification of 6.8B for reproductive toxicity is proposed.

Conclusion: Triggers classification of 6.8B for reproductive toxicity.

General conclusion about reproductive/developmental toxicity

The substance should be classified as 6.8B suspected of reproductive toxicity in animals (reduced

fertility in males).

Target Organ Systemic Toxicity [6.9]

Type of study: 13 week oral toxicity in the rat

Flag: Key study

Test Substance: Kasumin technical

Endpoints:

LOAEL: Males: 1000 ppm (equivalent to 58.2 mg/kg bw/day based on eosinophilic bodies in the

proximal tubular cells. Females 3000 ppm (equivalent to 395.5 mg/kg bw/day based on reduced body

weight gain and feed consumption (EPA staff considering the lung effects not be clearly treatment-

related).

NOAEL: 300ppm (equiv to 17.53 mg/kg bw/day in males) and 1000 ppm (equivalent to 69.2 mg/kg

bw/day in females) respectively.

42


September 2013

Reference: Nakashima, N.; 1991. Kasugamycin: 13 week oral sub-chronic toxicity study in rats. Study

No. IET 89-0083. Institute of Environmental Toxicology, Kodaira, Tokyo 187, Japan.

Klimisch Score: 2 [EPA Staff note the applicant has only supplied 41 pp of the study report, not the full

study report as is usually supplied, enabling verification of the study summary.]

Amendments/Deviations:

GLP: Yes

Test Guidelines: There is reference to compliance with MAFF, Japan (1985), EPA FIFRA (1984) and

OECD (1981) guidelines, below reference to GLP requirements, but the references are not precisely

drawn.

Species: Rats

Strain: Wistar

No/Sex/Group: 12

Dose: 0, 300, 1000, 3000 and 6000 ppm in diet. The intakes were equivalent to 0, 17.53, 58.2, 176.7

and 354.8 mg/kg bw/day (in males) and 20.33, 69.2, 201.0 and 395.5 mg/kg bw/day (in females).

Exposure Type: Oral by dietary administration.

Study Summary:

No clinical signs showed any statistically significant difference in incidence between any treated and

control groups.

No deaths occurred in any groups.

The 6000 ppm males consistently exhibited reduced body weight gains in comparison to controls from

Week 2. Females at this dose also showed reduced body weight gains and this was of statistical

significance in week 6.

The animals fed 3000 ppm and less had comparable body weights to controls.

Food consumption was reduced in 6000 ppm groups at weeks 1 and 4 in males and 1, 6 and 7 in

females and overall the food intake was reduced throughout treatment in these groups in comparison

to controls. In the 3000 ppm group the intake in week 1 were reduced, but there were not remarkable

differences in comparison to controls overall. The 1000 ppm group also had reduced intake in week 1,

but overall the intakes were not reduced, and at the lowest dose there was no difference.

At 6000 ppm there was a consistent increase in water consumption in males in most time period and

an increase in weeks 2 and 13 in females.

No changes were observed in the ophthalmological examinations in the 6000 ppm animals at 13

weeks.

The 6000 ppm animal showed low pH in urine and the females increased number of epithelial cells in

the urinary sediment. A decrease in urinary pH was seen only in females at 3000 ppm.

43


September 2013

Haematology

At 6000 ppm males and females had reduced haematocrit, haemoglobin and erythrocyte counts.

Females also showed an increase in the segmented form neutrophil count.

At 3000 and 1000 ppm, the males show reduced haematocrit, haemoglobin and erythrocyte counts.

At 3000 ppm the females had a decrease in mean corpuscular volume, although no dose dependency

was seen.

Blood biochemistry

At 6000 ppm groups (males and females) reduced glutamate pyruvic transaminase (GPT), total

protein (TP), albumin (Alb) and globulin were seen together with an increase in chlorine. In males a

decrease in calcium was seen.

At 3000 ppm , TP, Alb and Glob levels were reduced in males. GPT levels were reduced in both

sexes. Males also showed a decrease in alkaline phosphatase, although this was not dose-

dependent.

At 1000 ppm, decreases in GPT and Alb were seen in males. A decreased calcium level was also

seen, but was not dose dependent.

At 300 ppm decreased GPT and Alb were seen in males only.

There were no gross abnormalities.

At 6000ppm, there were increases in absolute and relative (to body weight EPA staff assumes)

cecum weight and increased relative kidney weights in females.

At 3000 ppm again absolute and relative cecum weight were increased, and relative kidney weights in

females only. The females also showed a decrease relative and absolute ovary weight, although no

dose dependence was shown.

At 1000 ppm males showed increase absolute cecum weight. Females showed reduced relative ovary

weight, with no dose dependency.

There were no changes to organ weight in the 300ppm groups.

Histopathology

At 1000 ppm and above the male proximal tubular cells of the kidney showed increased incidence of

eosinophilic bodies. Further staining did not identify these as lipofuscin deposits (as in previous

studies with the same study).

Females treated with 1000 ppm and more, the incidence of “foam cell aggregation” in the lung was

significantly increased. This finding was also seen in some female controls and the severity of the

lesions in the treatment groups were less than in controls.

The incidences of these histopathology lesions

Sex Organ Lesion Dose group (ppm):

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September 2013

- - - 0 300 1000 3000 6000

Male Kidney

Increase

eosinophilic

bodies.

0/12 0/12 7/12** 11/12** 11/12**

Female Lung Foam cell

aggregation 3/12 6/12 8/12* 11/12** 10/12**

*and ** statistically significant at the 5 and 1% confidence level respectively.

In the males at 1000 ppm there were also foam cell infiltration, but this was not dose-related.

There were no histopathological findings associated with the increased cecum or salivary gland

weights.

At 300 ppm there were no microscopic lesions with increased incidence in either sex.

Discussion

The reduction in body weight was attributed by authors to reduced food intake not reduced food

efficiency.

Increased water consumption is also considered treatment-related as similar findings were reported in

other studies.

The urinary parameter changes consistent with the micropathological findings are consistent with

kasugamycin being toxic to the kidney.

The blood parameter changes haematocrit, haemoglobin concentration and red blood cell counts

were considered treatment-related, although they were slight, because they were consistent and

dose-related. The segmented neutrophil changes in females were considered incidental, since such

changes have not been seen in other studies with the substance.

The primary target organ is the kidney. The blood biochemistry changes (reductions in GPT, TP, Alb

and globulin in males at 3000 and 6000 ppm and in females at 6000 ppm) are inter-related according

to the authors (total protein being influenced by globulin and albumen concentrations). However, in

the absence of histopathological liver changes the blood biochemistry findings of decreased albumin

without reductions in TP and globulin in males at 1000 and 300 ppm are not considered of

toxicological significance.

The pulmonary findings in females (and some males) while increased to a statistically significant

degree were not considered (by EPA staff) to be clearly treatment-related, since study author’s noted

in particular the background incidence of this finding. The authors suggest it is associated with

treatment with antibiotics based on other reports. The authors also suggest that the increased cecum

weight could relate to the antibiotic treatment of the animals.

45


September 2013

Conclusion: Based on the adverse effects in the kidney at 1000 ppm (equivalent to 58.2 mg/kg

bw/day in the male), the substance should be classified 6.9B for target organ toxicity.

See also the chronic/carcinogenicity study in the rat (reported above):

Non-neoplastic effects

LOAEL: 3000 ppm (equivalent to 116 and 139.8 mg/kg bw/day in males/females respectively)

based on organ weight and associated histopathological changes in the cecum and proximal

tubules of the kidney.

NOAEL: 300 ppm (equivalent to 11.31 and 13.42 mg/kg bw/day in males/females respectively)

Conclusion: EPA staff consider these data support the 13 week findings bearing mind the differing

dose spacing.

Type of study: 13 week toxicity study in mice


Test Substance: Kasumin technical (lot number KP-821)

Endpoints:

LOAEL: 3000 ppm ( equivalent to 408.5 and 565.8 mg/kg bw/day in males and females respectively)

based on mortality, perianal reddening, histopathological changes in the kidneys, testes and anus

NOAEL: 1000 ppm (equivalent to 135.4 and 170.9 mg/kg bw/day in males and females respectively)

Reference: Holmes P (1991) Kasugamycin: toxicity study by dietary administration to CD-1 mice for

13 weeks. LSR report no 90/HKC005/0345. Life Science Research Ltd, Eye, Suffolk IP23 7PX, UK.

Klimisch Score: 2 (Full study report was not provided)


GLP: Yes

Test Guidelines:

Species: Mouse

Strain: CD-1

No/Sex/Group: 12

Dose: 300, 1000, 3000 and 10000 ppm (equivalent to 0, 41.2, 135.4, 408.5 and 1559 mg/kg bw/day in

males and 0, 58.0, 170.9, 565.6 and 1834 mg/kg bw/day in females, respectively) [NB staff have

taken the dose values from the US EPA fact sheet as the full study report was not provided.]

Exposure Type: Oral by dietary administration.

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September 2013

Study Summary: Perianal reddening was observed from week 4 of treatment in males and females

receiving 3000 and 10000 ppm. Two males receiving 10000 ppm and one female receiving 3000 ppm

were killed on humane grounds in weeks 12 and 13 respectively because of extensive perianal skin

lesions. In addition, one female at 10000 ppm and one male and one female at 3000 ppm were found

dead in weeks 6, 7 and 14 respectively. There had been no history of ill health in these animals apart

from perianal reddening. Bronchopneumonia was noted at histopathological examination of the

female which had received 10000 ppm. Microscopic examination revealed active chronic inflammation

and ulceration of the anus in four of the six animals.

Females in the 10000 ppm group consumed slightly less food than their controls. Over the first 11

weeks of treatment, body weight gain was markedly reduced in males and females receiving 10000

ppm (62 and 58% of control values respectively). Males receiving 3000 ppm gained slightly more

body weight than controls while weight gain was unaffected in females given 3000 ppm and both

sexes in the 300 and 1000 ppm groups. Assessment of overall body weight gain (up to week 14) was

confounded by the effects on body weight of the clinical pathology assessments performed in weeks

12 and 13.

Haematology

Packed cell volume, haemoglobin concentration and erythrocyte count were reduced in males and

females given 10000 ppm, although only the changes in packed cell volume and haemoglobin

concentration in females were statistically significant. Neutrophil counts were slightly elevated in

males given 3000 and 10000 ppm. No effects were seen at lower doses.

Blood chemistry

After 12 weeks of treatment plasma total cholesterol and albumin levels were reduced in males and

females given 10000 ppm. The total cholesterol level was also reduced in females at 3000 ppm. No

toxicologically significant effects were seen at lower doses.

Histopathology

There was a dose-dependent effect on the pars recta region of the proximal tubule of the kidney in

females given 3000 and 10000 ppm. This consisted of a diffuse basophilia and hyperplasia of the

tubular epithelium, but this was not associated with any epithelial degeneration or necrosis. In the

absence of necrosis the authors considered it was unclear whether these changes were an adaptive

or regenerative response, and the toxicological significance of these changes was considered to be

uncertain.

In the testes, seminiferous tubule dilatation and degeneration was observed in males given 10000

ppm, and in one male administered 3000 ppm.

Males and females given 3000 or 10000 ppm were found to have ulceration and inflammation of the

anus. One female receiving 1000 ppm had slight anal inflammation but no ulceration.

Conclusion: The substance does not require classification for target organ toxicity based on the

results of this study.

47


September 2013

Type of Study: 13 week repeat dose study in dogs


Test Substance: Kasugamycin hydrochloride technical (80.6% pure)

Endpoints:

LOAEL: Study authors proposed 3000 ppm as the LOAEL based on the microscopic findings in the

tongue. EPA staff consider that 300 ppm (10.59 mg/kg bw/day in males) should be defined as the

LOAEL based on the reddening of the oral cavity.

NOAEL: 300 ppm was claimed by the study authors, but this ignores the reddening of the oral cavity

seen in one male at 300ppm, which could be seen as the first indication of a toxicological finding. EPA

staff consider that no reliable NOAEL has been established in this study in males, but 11.44 mg/kg

bw/day could be considered a NOAEL in females.

Reference: Thomford, P. J.; 1993. 13 –Week Dietary Toxicity Study in Kasugamycin in Dogs.

[Laboratory Project Identification HWI 6434-101] Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard,

Madison, Wisconsin 53704, USA.

Klimisch Score: 1

Amendments/Deviations: EPA staff note the change to the dose levels during the study and the use of

canned dog food in the Group 4 animals during week 7.

GLP: Yes

Test Guidelines: FIFRA Guideline 82-1

Species: Dog

Strain: Beagle

No/Sex/Group: 4

Dose levels: 0, 300, 3000 and 6000 ppm. The animals in the 6000 ppm group were fed this diet

through Day 1 – Day 41, but because of decreased food consumption, body weight loss and tongue

lesions the animals were taken off the test material through Day 49. On Day 50 the animals were

offered diet containing 4500 ppm. The animals remained on this diet for the remainder of the study.

The dogs were treated with kasugamycin for at least 12 weeks during the study.

The above dietary concentrations gave rise to intakes as follows:

Males: 300 (10.59 mg/kg bw/day), 3000 (105.96 mg/kg bw/day), 4500 [Weeks 8 – 13], 157.14 mg/kg

bw/day and 6000 [Weeks 1 – 5] (212.1 mg/kg bw/day.

Females: 300 (11.44 mg/kg bw/day), 3000 (107.89 mg/kg bw/day), 4500 [Weeks 8 – 13], 172.93

mg/kg bw/day and 6000 [Weeks 1 – 5] (185.33 mg/kg bw/day).

48


September 2013


Study Summary:

In the range finding study over 4 weeks, 3 dogs per sex/group were fed diets with 0, 300, 1000, 3000,

10,000 and 30,000ppm. Decreased palatability caused decreasing body weight and food consumption

was observed. On Day 20 the animals in the 30,000 ppm group were sacrificed. One animal from the

10,000 ppm group was sacrificed on Day 26.

In the main study, the dose groups were 0, 300, 3000 and 6000 ppm but the top dose underwent a

period of withdrawn treatment due to decreased feed consumption before being dosed at 4500 ppm.

Observations in group 4 (3000 ppm) included discoloured and few faeces, swollen mouth, excessive

salivation and tongue lesions (beginning on Day 35). At approximately Day 50 males and females in

group 3 were also observed with tongue lesions. One female was seen with tongue lesions beginning

on Day 37. Since food consumption was not decreased substantially the group 3 animals were

retained at 3000 ppm. Excessive salivation was seen in females in group two (300 ppm).

There were no ophthalmological abnormalities.

Clinical pathology revealed mildly lower alkaline phosphatase for male and females in Group 4,

attributed by the study authors to reduced food consumption and weight loss. There was midly to

moderately lower cholesterol for animals in Groups 3 and 4, the study authors did not attempt an

explanation noting the use of canned dog food in Week 7 for Group 4 animals.

Terminal body weights were lower for the animals in Groups 3 and 4.

Findings at necropsy included erosions/ulceration of the tongue in Group 3 and 4 animals, diffuse

reddening for one females in Group 4 and a depressed area for one male in Group 3. There was

thickening of the skin of the commissure of the mouth for animals in Groups 3 and 54 and diffuse

reddening of the oral cavity for one male in Group 2, one male and two females in Group 3 and one

female in Group 4.

There were not test material related changes in absolute organ weights, organ to body weight

percentages or organs to brain weight ratios.

Microscopically the studies targeted the tissue of the tongue. There was a consistent progression.

Initially loss of the papillae of the dorsal epithelium occurred, progressing from the lateral to the centre

of the tongue. Epithelium then atrophied until it was missing causing ulceration. Ulcerated areas

frequently contained serous fluid and minimal to slight chronic-active inflammation. No findings in the

tongue were seen in Group 1 (control) or Group 2.

Conclusion: Assuming the reddening of the oral cavity in one male animal is considered an adverse

effect, the classification supported by this study is 6.9A. [EPA staff note that the 52 week dog data

should be considered before reaching a final view, and in the subsequent 52 week study lower toxicity

was demonstrated.]

49


September 2013

Type of Study: 52 week repeat dose study in dogs


Test Substance: Kasugamycin hydrochloride technical (80.6% pure)

Endpoints:

LOAEL: >3000 ppm (>99.6 mg/kg bw/day in males and >103.6 mg/kg bw/day in females)

NOAEL: In this study the NOAEL is proposed at 3000 ppm in dogs (99.6 mg/kg bw/day in males and

103.6 mg/kg bw/day in females. EPA staff note that based on these finding the outcome of the 13

week study can be disregarded.

Reference: Albretsen, J. C.; 2003. 52 –Week Dietary Toxicity Study with Kasugamycin in Dogs.

[Covance Study Number 6434-117] Covance Laboratories Inc. 3301 Kinsman Boulevard, Madison,

Wisconsin 53704 - 2595, USA.

Klimisch Score: 1


GLP: Yes

Test Guidelines: US EPA OPPTS 870.4100 (1998). OECD No 452 (1981)

Species: Dog

Strain: Beagle

No/Sex/Group: 4

Dose levels: 0, 300, 1000, and 3000 ppm equivalent to 0, 10.5, 30.5 and 99.6 mg/kg bw for males and

9.4, 33.4 and 103.6 mg/kg bw/day in females.


Study Summary:

The study authors took into account the findings of the 13 weeks study which suggested that tongue

lesions may be expected and that a dose level of 4500 ppm would be higher than could be tolerated.

Therefore the dose levels selected were 0,300, 1000, and 3000 ppm for this 1 year study.

One female dosed at 3000ppm died on Day 14. The cause of death was not able to be determined,

although she had no formed faeces on Days 10 and 13, and a red discharge was visible in the cage

pan on the day of her death. The death was not considered to be treatment related.

One animals (a female in 300 ppm), developed a swollen muzzle beginning on day 346, which

increase and began to involve areas around the head in addition. The veterinarian injected

antihistamine and the swelling subsided and by Day 352 was no longer observed. The study authors

50


September 2013

consider this is likely to have been an allergic reaction but not to the test substance, since she had

been exposed to that for 300 day prior to the first observation.

There were no test-substance related clinical or ophthalmic observations of effects on physical

examination body weight, body weight changes, food consumption or organ weights. There were no

test article related macroscopic or microscopic findings.

No abnormalities in ophthalmic findings, body weight differences or food consumption or food

efficiency were identified.

Administration of 3000 ppm was associated with minimally higher urea nitrogen and creatinine, lower

urine volume and higher urine specific gravity.

Based on these findings the NOAEL for the substance in dogs for 52 weeks is 3000 ppm.

Conclusion: Based on these data no classification for repeat dose target organ toxicity via the oral

route should apply to the substance.

Type of study: Repeat dermal exposure over 21 days in the rat



Endpoints:

Systemic

LOAEL: > 500 mg/kg bw/day (males) and > 200 mg/kg bw/day (females

NOAEL: 500 mg/kg bw/day (males) and 200 mg/kg bw/day (females

Dermal

LOAEL: 500 mg/kg bw/day (males) and 200 mg/kg bw/day (females

NOAEL: 250 mg/kg bw/day (males) and 100 mg/kg bw/day (females (expressed as the active

ingredient, free base)

Reference: Seidel, S.D.; 2009. A 21-day dermal toxicity study of Kasugamycin technical in Sprague

Dawley rats. Study No. WIL-476005. WIL Research Laboratories, LLC, 1407 George Road, Ashland,

OH 44805-8946, USA.

Klimisch Score: 1


GLP: Yes

Test Guidelines: US EPA OPPTS 870.3200

Species: Rats

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September 2013

Strain: Crl: CD (SD)

No/Sex/Group: 10

Dose: 0, 50, 250, and 500 mg/kg bw/day respectively, but beginning on Day 8 the dose levels for the

two top dose, female groups were reduced from 250 and 500 mg/kg bw/day to 100 and 200 mg/kg

bw/day respectively, due to the severity of the dermal observations.

Exposure Type: Dermal patch for 6 hours per day for 21 days.

Study Summary: There were no test substance-related deaths or clinical observations related to

systemic toxicity. Body weights and food consumption were unaffected by test substance.

There were no test substance-related microscopic findings at the dermal application site of the 50 and

250 mg/kg bw/day group males or the 50 and 100 mg/kg bw/day group females.

Clinical observations of dose site reddened and scabbing within the dose site were noted in the 250

and 500 mg/kg bw/day group males and females beginning on study day 7 and 5 for males and

females, respectively. These observations generally persisted throughout the dosing period and

continued after the dosage level reduction for females.

Dermal observations included erythema, oedema, eschar, coriaceousness, pinpoint scabbing, and

encrustation in the 250 and 500 mg/kg bw/day group males and females. On study day 7, 40% and

90% of females in the 250 and 500 mg/kg bw/day groups, respectively, were noted with very slight to

severe erythema and/or oedema in a dose-dependent manner, with eschar and coriaceousness noted

in the 500 mg/kg bw/day group females. As a result of these effects, the dosage level for the 250 and

500 mg/kg bw/day group females was reduced to 100 and 200 mg/kg bw/day, respectively, on study

day 8. Following the decrease in dosage, a reduction in the number and severity of dermal

observations was noted in the 250/100 mg/kg bw/day group females while dermal observations in the

500/200 mg/kg bw/day group females continued to vary in severity. In the 500 mg/kg bw/day group

males, 40% of the animals were noted with very slight to slight erythema and/or oedema on study day

7 with observations progressing in incidence and severity over the course of the dosing period to

result in 80% of the 500 mg/kg/day group males being noted with very slight to severe erythema

and/or oedema, along with observations of eschar and encrustation, on study day 21. A single 250

mg/kg bw/day group male was noted with test substance-related very slight erythema on study day 7

which had resolved by the next observation period on study day 14.

Macroscopic necropsy observation of scabbing were noted in 4/10 males from the 500 mg/kg/day

group and 2/10 females from the 200 mg/kg bw/day group. These 2 females from the 200 mg/kg

bw/day group also had open sores on the test substance application site.

Nine of 10 males from the 500 mg/kg bw/day group and 7/10 females from the 200 mg/kg bw/day

group had microscopic findings of acanthosis, acute inflammation, and/or ulceration affecting the test

substance-treated skin. Two females from the 200 mg/kg bw/day group also had minimal

granulomatous inflammation. The lesions were focal or segmental and usually affected only 1 of the

examined tissue sections from the test substance-treated skin site. Ulceration was accompanied by

52


September 2013

acute inflammation composed of infiltrating and exuding neutrophils. Acanthosis was usually adjacent

to the areas of ulceration.

While not directly test substance-related, stress responses consisting of mild myeloid hyperplasia in

the sternal and femoral bone marrow or minimal lymphoid necrosis of the thymic cortex and minimal

vacuolation of the adrenal cortex were noted in 2 males from the 500 mg/kg/day group and minimal

lymphoid necrosis was noted in the thymus of 2 females from the 200 mg/kg/day group and 1 control

group female. These alterations were considered to be secondary to dermal inflammation or stress

responses as part of the dose application and handling procedures.

There were a number of deaths during the study which were not substance-related. One male control

animal on study Day 2, one male in the 50 mg/kg bw/day group on Day 5, and a female from the 100

mg/kg bw/day dose group on Day 10. It appears from the description from the authors that two of

these deaths related to wrapping procedure and handling of the animals causing respiratory distress.

Conclusion: The substance does not trigger target organ toxicity for repeat exposure via the dermal

route.

Comments on classification: EPA staff note that the adverse effects seen at doses at the

thresholds in table 17.2 (User Guide p17-9) were non systemic effects at site of exposure, and such

findings should not be identified as significant toxicological effects for systemic target organ toxicity

from the dermal route.

Type of Study: Acute neurotoxicity study in rats

Flag: Key study

Test Substance: Kasugamycin

Endpoints: Neurotoxicity

LOAEL: > 2000 mg/kg bw

NOAEL: 2000 mg/kg bw

Reference: Beck, M. J.; 2009 An oral (gavage) acute neurotoxicity study of Kasugamycin technical in

rats. Study No. WIL-476009. WIL Research Laboratories, LLC, 1407 George Road, Ashland, OH

44805-8946, USA.

Klimisch Score: 1


GLP: Yes

Test Guidelines: OECD No 424. US EPA OPPTS 870.6200.

Species: Rats


53


September 2013

No/Sex/Group: 12

Dose: 0 500, 1000 and 2000 mg/kg bw

Exposure Type: Oral gavage (single dose)

Study Summary: There were no reported findings of relevance in a study using a neurotoxicity

screening battery consisting of a functional observational battery, a locomotor activity and a

neuropathological assessment.

Conclusion: The substance did not demonstrate any ability to cause acute neurotoxicity in the study.

Type of Study: Repeat dose (90 day) neurotoxicity in the rat

Flag: Key study


Endpoints:

LOAEL: 6000ppm (439 mg/kg bw/day in males) and 486 mg/kg bw/day in females)

NOAEL: >6000ppm (439 mg/kg bw/day in males and 486 mg/kg bw/day in females) [no subchronic

neurotoxicity was observed in the study].

Reference: Beck M. J.; 2009. A 90-day Dietary Neurotoxicity Study of Kasugamycin Technical in

Rats. (Study Number WIL-476008. WIL Research Laboratories, 1407 George Road, Ashland, OH

44805-8946, Ohio, USA.

Klimisch Score: 1

Amendments/Deviations: None [The study Director, Beck M. J. replaced the initial study Director

Roegge, C. S. on 14 August 2009 (the study having started on 12 May 2009].

GLP: Yes

Test Guidelines: OECD No 424. US EPA OPPTS 870.6200

Species: Rat


No/Sex/Group: 12

Dose: 0, 300, 3000, 6000 ppm in diet, equivalent to intakes of 0, 21, 210 and 439 mg/kg bw/day in

males and 0-, 23, 238 and 486 mg/kg bw/day in females respectively.


Study Summary:

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September 2013

Test substance related effects were limited to reduced mean body weight and body weight gain in the

6000 ppm group animals. There were no test substance related effects in the 300 or 3000 ppm

groups.

There were no effects observed on home cage observation, or handling.

Open field observation showed a possible effect (reduction) on the mean numbers of grooming counts

for the 300 and 3000 ppm group females compared to controls, at study week 3 and 7 , and at 6000

ppm group females in study week 1, 3, and 7 evaluations. However, the grooming counts were

generally within the WIL historical control ranges and the magnitude of the changes were not

considered by authors to be toxicologically relevant.

There were no statistically significant differences in the sensory observations.

There were no test substance related effects on neuromuscular observations.

Locomotor activity patterns were unaffected by test substance. An increase in ambulatory counts was

seen for 6000 ppm females in the overall session at the study week 12 evaluation in comparison to

controls. However, the same parameter had been raised (to a non-statistically significant degree) in

the pre-test evaluation. No remarkable shifts in the pattern of habituation occurred in any of the test

substance-exposed groups. The changes were not attributed to the test substance by the authors.

Pathological examination revealed no differences in brain weights, and no test substance-related

findings in the central or peripheral nervous system.

Conclusion: The substance does not demonstrate neurotoxicity after repeat oral administration.

General conclusion about target organ systemic toxicity

The data from the 13 week rat study showing kidney effects including pathological findings in males at

58.2 mg/kg bw/day support classification for systemic toxicity following repeat exposure via the oral

route. The chronic study in rats has dose spacing which precludes a firm conclusion that this is not

justified. Nevertheless, it is noted that no such classification is justified based on the data for dog

(particularly the 1 year study).

The kidney was identified as a target organ in both rats and mice. Testicular effects, consistent with

the findings from the reproductive toxicity study, were also observed in rodents.

Other studies

Metabolism studies

Type of Study: Metabolism in the rat of 14

C labelled kasugamycin

Flag: Key study

Test Substance: 14

C labeled Kasugamycin [The label is a single carbon at the methyl group

hexopyranose ring.]

55


September 2013

Reference: Cheng T, 1998. Metabolism of 14

C-kasugamycin in the rat. Covance Study No: 6434-110.

Covance Laboratories Inc. 3301 Kinsman Boulevard, Madison, Wisconsin 53704, USA.

Klimisch Score: 1

Amendments/Deviations

GLP: Yes

Test Guidelines: US FIFRA 40 CRF 158, Series 85-1. OECD No 417.

Species: Rat

Strain: Fischer 344

No/Sex/Group: 2-9 (see below)

Dose: The study has a complicated design with the following groups (number of males and females

respectively in brackets):

A: Oral low dose (100 mg/kg bw) (5/5)

B: Repeat group (100 mg/kg bw). (Radiolabel introduced on the last day of a 14 day exposure). (5/5)

C: Oral High dose (1000 mg/kg bw) (5/5)

D: Repeat dose (1000 mg/kg bw/day) (Radiolabel introduced on the last day of a 14 day exposure).

(5/5)

E: Pharmacokinetics Low dose (100 mg/kg bw) (4/4)

F: Pharmacokinetics High dose (1000 mg/kg bw) (4/4)

G: Toxicodynamics Low dose (100 mg/kg bw) (9/9)

H: Toxicodynamics High dose (100 mg/kg bw) (9/9)

I: Bile low dose (100 mg/kg bw) (4/4)

J: Bile high dose (100 mg/kg bw) (4/4)

K: Control (2/2)

Exposure Type:

Study Summary:

Following administration of an oral dose of 14C –kasugamycin at 100 or 1000 mg/kg bw the total

radioactivity recovery ranged from 90.6 to 96.7%, with 87.7 to 94.5% eliminated in faeces, and 1.35 to

3.26% ion urine. In the preliminary study less that 0.2% of the radioactive was exhaled as 14

CO2.

The Cmax was reached approximately 1 hour after administration with peak plasma Cmax of 1.47

and 2.17 ppm (for male and females respectively – low dose, and 6.40 and 5.23 ppm (for male and

females respectively high dose). The terminal elimination half-lives were 1.41 and 1.17 hours for

56


September 2013

Group E males and females respectively and 1.40 and 1.55 hours for Group F males and females

respectively.

Kidney had the highest tissue residue levels at all time points, declining to 3 -4ppm (low dose) and 24-

25 ppm (high dose) at 168 hours post-dose. Lymph nodes also had higher residue concentrations

than plasma, but these declined to non-detectable by 168 hours post-dose.

Regardless of the dose regime, less than 5% of the oral dose was absorbed. Nearly all the dose

administered (>90%) was eliminated in faeces as the parent compound. A portion of the absorbed

kasugamycin was eliminated in urine as the unchanged parent compound, while a small amount

(<0.2%) of the total kasugamycin dose was metabolized to kasugamycinic acid and

kasuganobiosamine through deamination, oxidation, decarboxylation and hydrolysis.

Conclusion: Kasugamycin is very poorly absorbed after oral administration (<5% in rats), and as may

be expected for a cardiac glycoside the kidney is a key target organ.

General conclusion about mammalian toxicology of active ingredient(s) and metabolite(s)

Table 3: Summary of studies with NOAEL and LOAEL values and key effects.

Study type NOAEL LOAEL Key effect

13 week oral toxicity in

the rat

300 ppm (equivalent to

17.53 mg/kg bw/day) in

males

1000 ppm (equivalent

to 69.2 mg/kg bw/day)

in females



in males



in females

Eosinophilic bodies in the

proximal tubular cells of

the kidney (males only).

Reduced body weight

gain and feed

consumption (females)

[EPA staff accepted the

lung effects not being

dose-related.]

[Bold NOAEL used to

derive AOEL.]

Chronic toxicity in the

rat

300 ppm (equivalent to

11.31 mg/kg bw/day

and 13.42 mg/kg

bw/day in

males/females

respectively)


to 116 and 139.8 mg/kg

bw/day in

males/females

respectively)

Based on organ weight

and associated

histopathological changes

in the cecum and

proximal tubules of the

kidney and testicular

atrophy.

[Bold NOAEL used to

derive ADE.]

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September 2013

13 week oral toxicity

study in the dog

EPA staff consider no

NOAEL established,

but authors propose

300 ppm (10.59 mg/kg

bw/day in males)

EPA stuff propose 300

ppm (10.59 mg/kg

bw/day in males)

Epithelial damage to the

tongue and other

changes in the buccal

cavity.


study study in the dog

3000 ppm in dogs (99.6

mg/kg bw/day in males

and 103.6 mg/kg

bw/day in females).

>3000 ppm (>99.6

mg/kg bw/day in males

and >103.6 mg/kg

bw/day in females)

No toxicologically

significant findings

identified.


study in mice


to 135.4 mg/kg bw/day

in males and 170.9

mg/kg bw/day in

females).


to 408.5 mg/kg bw/day

in males and 565.6

mg/kg bw/day in

females).

Mortality, perianal

reddening,

histopathological changes

in the kidneys, testes and

anus.

78 carcinogenicity

study in mice

1500ppm (equivalent

to186.3 mg/kg bw/day

and 215.2 mg/kg

bw/day in females).

>1500ppm (equivalent

to >186.3 mg/kg

bw/day and >215.2

mg/kg bw/day in

females).

1500 ppm in the male

was initially identified as a

LOAEL, based on a

reduction in the incidence

of enlarged spleen, but

this is not considered a

toxicological significant

finding.

Developmental study in

rodents

200 mg/kg bw/day in

maternal animals

1000 mg/kg bw/day for

fetal animals.

1000 mg/kg bw/day for

maternal animals

>1000 mg/kg bw/day

for fetal animals.

Reduced feed intake in

maternal animals.

No adverse foetal effects.

2 generation study in

rat

Parental toxicity: 1000

ppm (equivalent to

69.99 and 84.86 mgkg

bw/day in males and

females respectively)

Reproductive toxicity:

1000 ppm in males

6000 ppm (421.8 mg/kg

bw/day in males and

505.17 mg/kg bw/day in

females)

6000 ppm (421.8 mg/kg

bw/day) in males

Red swollen skin around

the anal opening and

macroscopic/microscopic

ulceration at the

boundary of the rectum

and anus (at necropsy).

Reduced fertility (males)

58


September 2013

Environmental fate - Robust study summaries for the active ingredient

and metabolites

The study reports about kasugamycin monohydrochloride monohydrate, also called kasugamycin

technical or kasugamycin were evaluated and summarised in the following section. When not

otherwise specified, kasugamycin means kasugamycin monohydrochloride monohydrate.

8.2.1. Water compartment

Ready Biodegradation

Not provided but a degradation study in water/sediment systems is available. So the requirement can

be waived.

Hydrolysis

Flag: Key study

Test substance: [hexopyranosyl-U-14

C]-kasugamycin hydrochloride hydrate, also called (14

C)-

kasugamycin

Endpoint: DT50

Value: more than 1 year at pH 4 and 5. DT50 = 129 days at pH 7 and 20°C and 11 days at pH 9 and

25°C.

Reference: S. Swales – (14

C)- kasugamycin: Hydrolytic stability. Study 1442/21 (21/05/2003)

Klimisch score: 1

Amendments/Deviations: No

GLP: Yes

Test guidelines: EEC method C7 (1994) ; Japan MAFF test guidelines (2000) ; EPA pesticide

assessment guidelines subdivision N, section 161-1 (1982).

Dose levels: 5 ppm

Analytical measurements: HPLC and TLC on selected extracts

Study summary: The hydrolytic stability of (14

C)- kasugamycin has been studied at pH 4 (25 and

50°C); pH 5 (25°C); pH 7 (25, 50, 62 and 74°C) and pH9 (25 and 50°C) in sterile aqueous solutions.

(14

C)- kasugamycin was applied to glass vials containing buffer (3 mL) in sterile water (15-34 µL) to

achieve final concentration of 5 ppm.

Radioactivity present in solution at the time of analysis was determined by liquid scintillation counting

(LSC). There was no decrease over the incubation periods and the applied radioactivity (>91% in

each sample) was in solution.

Samples were analysed directly for (14

C)- kasugamycin and hydrolysis products by HPLC and by TLC

to clarify degradation product that was present.

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September 2013

Half-life values were computed by regression analysis assuming first order kinetics. The Arrhenius

equation was used to compute half-life values at pH 7 at environmentally relevant temperatures (20°C

and 25°C).

Kasugamycin was shown to degrade at the following rates:

pH Temperature (°C) DT50 (Days) DT90 (Days)

4 25 580 1948

5 25 658 2233

7

25 72.4 253

50 3.57 12.3

62 1.57 5.50

74 0.51 1.79

9

25 10.8 37.2

50 7.28 hours 25.5 hours

The rates of degradation at pH 4 and 5 and 25°C were similar and more than 1 year. The degradation

rate increases with increasing pH values: at pH 7, the calculated half-life at 20°C is 129 days. At pH 9,

the DT50 is 11 days at 25°C.

At all pH values studied, there was only one major degradation product present, and this was

identified as kasugamycinic acid. The highest levels of kasugamycinic acid were found at pH 9 (up to

78% of the applied radioactivity at 30 days, end of the study) with the lowest levels being found at pH

4 and 5.

Conclusion: Hydrolysis will not contribute significantly to kasugamycin degradation in

environmental conditions except at high pH.

Aqueous Photolysis

Flag: key study



C)-

kasugamycin

Endpoint: DT50

Value: 8.2 days in natural lake water and 436 days in pH 5 buffer

Reference: (14

-C)-kasugamycin: photodegradation in sterile aqueous solution. L. Bishop & L.

Maudsley. Study number 1442/22- June 2003

Klimisch score: 1

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September 2013

Amendments/Deviations: no deviations

GLP: yes

Test guidelines: Japan MAFF test guidelines section 2-6-2 (2001); SETAC procedures for assessing

the Environmental fate and ecotoxicity of pesticides, section 10 (1995); EPA pesticide assessment

guideline, subdivision N section 161-2 (1982)

Dose levels: 5 ppm

Analytical measurements: liquid scintillation counting, HPLC, TLC and LC/MS/MS for confirmatory

purposes

Study summary: the photodegradation of (14

-C)-kasugamycin was studied in pH5 buffered solution

and natural lake-water under sterile conditions at 25 ± 1oC over a period of 18.9 days. The study was

performed to assess the rate of degradation of the test substance at the pH of greatest hydrolytic

stability and in natural lake water.

Throughout the study > 92% of applied radioactivity was recovered from irradiated and dark control

samples at each time point.

A maximum of 2% of the radioactivity was found in the traps for volatile compounds. This does

indicate that some mineralization must have occurred.

One major degradation product was present, and this was identified as kasugamycinic acid. Levels

were highest in irradiated lake water (55.6% of applied radioactivity after 12.9 days. This degradation

product was also found at lower levels in the dark control lake-water samples (21.5% after 18.9 days).

In pH 5 buffer, kasugamycinic acid levels were much lower even in irradiated samples (3.6% after

18.9 days) and a maximum of 1.4% of the compound was detected in dark-control buffer samples.

Other degradation products were also formed in the irradiated samples. These included

kasuganobiosamine (4.7% of applied radioactivity after 12.9 days) and several minor unidentified

degradation products. These were most abundant in the irradiated lake-water samples.

The DT50 (calculated with first order kinetics and the characteristics of a Japanese spring day) with

light exposure in natural lake water was 14 days and the DT90 was 44 days. The DT50 with the light

exposure in buffered solution at pH 5 was 260 days and the DT90 was 875 days.

The Canadian PMRA has re-calculated the DT50. DT50 (single first order) = 436 d for sterile buffer and

8.2 days in lake water. (PMRA Kasugamycin Proposed Registration decision, 27/12/2012 PRD2012-

30). These values will be considered for our risk assessment

Conclusion: DT50 = 8.2 days in natural water

Water/sediment aerobic biodegradation

Flag: key study

Test substance: [D-glucosamine-U-14

C] kasugamycin

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September 2013

Endpoint: DT50 (whole system)

Value: 18.2 and 28.6 days

Reference: Aerobic aquatic soil metabolism of [14

-C] kasugamycin. K.Shepler. Study number 1819W.

26/10/2009

Klimisch score: 1

Amendments/Deviations: no

GLP: yes

Test guidelines: OECD 308

Dose levels: 4.16 mg/kg dry sediment

Analytical measurements: HPLC, liquid scintillation counting, TLC

Study summary: An aerobic metabolism study was conducted with [14

-C] kasugamycin applied to two

sediment/water systems freshly collected from Golden Lake (loamy sand, water pH=8.1) and Goose

River (clay loam, water pH=7.8) in North Dakota. Individual samples were treated with [14

-C]

kasugamycin and incubated in the dark under aerobic conditions at 25 ± 1oC for approximately 100

days. The application rate was 4.16 mg a.i./kg dry sediment for both test systems. For each test

system, sediment/water samples were collected in duplicate at six time points for duration of the study

(100 days).

The samples were continuously aerated throughout the incubation period. The water and sediment

extracts were quantified by liquid scintillation counting (LSC). Kasugamycin and its metabolites were

quantified by HPLC of water phase and sediment extracts. The assignment of peaks in the primary

chromatography was confirmed using 2-dimensional thin layer chromatography.

Material balance based on the sum of water layers, sediment extracts, bound residues and trapped

volatiles averaged 92.2 ± 5.1% of applied radioactivity (AR) for the Golden Lake system and 92.4 ±

6.2% AR for the Goose River system.

Extractable radiocarbon from the soil represented an average of 1.0% AR and 1.5% at t0 in Golden

Lake and Goose River systems, respectively, increasing to a maximum average of 38.0% AR and

43.8%AR after 14 days and subsequently declining to an average of 21.4%AR and 23.5%AR (Golden

Lake and Goose River, respectively) following 100 days of incubation.

The radiocarbon present in the water layer represented > 95% AR at t0 in both test systems, declining

to less than 3%AR after 100 days.

Non-extracted residues represented <0.5% AR at t0, increasing to an average of 17.8% AR (Golden

Lake) and 18.3% AR (Goose River) at the end of the study. Radiocarbon in the organic volatiles traps

represented negligible amounts of radiocarbon (< 0.2% AR) throughout the study, while up to 44.1%

AR (Golden Lake) and 43.7% AR (Goose River) were recovered as CO2.

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September 2013

Kasugamycin degraded moderately fast in both test systems throughout the study period. Parent

substance represented an average of 21.3% AR (Golden Lake) and 20.9% (Goose River) at the end

of the study. Kasugamycinic acid was the major degradate detected in both systems and represented

an average of 28.4% AR (Golden Lake) and 15.9% AR (Goose River) by day 14, decreasing to 1%

AR at the end of the study. All other degradates were individually < 5.8% AR in both systems.

The degradation rate of kasugamycin under aerobic conditions was determined using first order

kinetics. The DT50 was calculated as 47.7 days with a correlation coefficient r2 of 0.75 for the Golden

Lake test system and 45.8 days with a correlation coefficient r2 of 0.81 for the Goose River test

system. The corresponding DT90 were 158.4 and 152.1 days, respectively. The rate of degradation for

the water layers was also calculated using first order kinetics. The DT50 was calculated as 6.1 days

with a correlation coefficient r2 of >0.95 for the Golden Lake test system and 7.4 days with a

correlation coefficient r2 of >0.95 for the Goose River test system.

The Canadian PMRA has re-calculated the DT50 and DT90: whole system Golden Lake DT50 = 18.2 d

(indeterminate order rate equation), DT90 = 179 d; for Goose River, the corresponding values are 28.6

and 95 days according to single 1rst order, respectively. (PMRA Kasugamycin Proposed Registration

decision, 27/12/2012 PRD2012-30). These values will be considered for our risk assessment.

Conclusion: DT50 (whole system) = 18.2 and 28.6 days

Water/sediment anaerobic biodegradation

Flag: key study

Test substance: 14

C-(U) –hexopyranosyl -Kasugamycin hydrochloride hydrate

Endpoint: DT50

Value: 170 days according to PestDF calculation tool for single first order (value calculated by the

applicant is 105 days)

Reference: Anaerobic aquatic metabolism of 14

C-kasugamycin on a representative agricultural soil. R

Fathulla and A Conteh. Study number 6434-108. 23/11/1998

Klimisch score: 2 (results not clearly reported in the report)

Amendments/Deviations: none

GLP: yes

Test guidelines: FIFRA subdivision N, section 162-3

Dose levels: 5.34 µg a.i./ g sample

Analytical measurements: radioactivity measured by liquid scintillation counting (LSC), identification of

degradation products by thin layer chromatography (TLC)

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September 2013

Study summary: An anaerobic aquatic soil metabolism study was conducted with 14

C-kasugamycin on

clay loam soil and well water in a water/sediment system. The anaerobic conditions were maintained

and checked during 50 days prior to fortification with the test item at 5.34 µg a.i./g. The incubation

conditions were: 25oC in a chamber flushed with nitrogen. Duplicate samples were removed after day

0, 3, 7, 14, 32, 63, 93, 122, 185, 277 and 368 days. At each interval, water was decanted and the soil

was extracted, the radioactivity was analyzed by LSC.

The results indicate that anaerobic conditions were maintained during the course of the study. The

mean total material balance ranged between 94.5 and 103.7%.

Kasugamycin was the major component of the radioactivity. Kasugamycinic acid (up to 33.6%),

kasuganobiosamine (up to 43.2%) and 2 other unidentified metabolites (up to 8.1 and 5.1%) were

also detected in the system. Bound residues accounted for up to 10.1 %.

It shows that kasugamycin breaks down through hydrolysis of the imine group to form kasugamycinic

acid and reductive cleavage of carboxyiminomethyl to form kasuganobiosamine.

The degradation rate of kasugamycin in anaerobic conditions is slow (DT50 of 170 days according to

single first order, recalculated by EPA staff).

Conclusion: DT50 = 170 days

Bioaccumulation potential

Not provided however based on the log Kow (< -1.96) value, it can be concluded that kasugamycin is

not potentially bioaccumulative. So no further study is necessary.

Soil compartment

Aerobic Degradation in Soil (routes and rates) – Laboratory studies

Flag: key study



C)-

kasugamycin

Endpoint: DT50

Value: 40.8 days (first order). The Pest Management Regulatory Agency (Canada) re-calculated the

DT50 with an indeterminate order rate equation. The DT50 is 39.5 days in these conditions (Ref:

Kasugamycin: Proposed registration decision PRD2012-30; 27/11/2012). As both values are very

similar, the applicant’s one was taken into consideration for the risk assessment (worst case).

Reference: Aerobic metabolism of 14

C-kasugamycin on a representative agricultural soil. R. Fathulla &

A. Conteh Study number 6434-106. 23/11/1998

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September 2013

Klimisch score: 2 Only 1 soil was included in this study but it is performed according to a validated

guideline and to GLP

Amendments/Deviations: Only 1 soil was included in this study.

GLP: yes

Test guidelines: FIFRA 40 CFR 158 Pesticide assessment guidelines, subdivision N-162-1

Dose levels: 5 mg ai/kg soil

Analytical measurements: Liquid scintillation counting (LSC) and thin layer chromatography (TLC)

Study summary: An aerobic soil metabolism study was conducted with 14

C-kasugamycin on clay loam

soil (pH 7.025; 2.3% organic matter). The samples were prepared by placing approximately 50 g dry

weight of soil in amber jars and the moisture content of the soil adjusted to approx. 75% of the field

moisture capacity. The samples were fortified with the test substance at a nominal concentration of 5

mg/kg dry soil. The samples were placed in a dark room at 25°C ± 1°C in chambers connected to a

series of volatile component traps for up to 1 year.

The total amounts of radioactivity in the extracts and the volatile traps were determined by liquid

scintillation counting (LSC). Further characterization of radioactivity was performed on aliquots of soil

samples with applied radioactivity (AR) up to 10% or more. These characterizations involved isolation

of humic acid, fulvic acid, and humin fractions of soil organic matter and determining the amount of

radioactivity associated in each fraction by LSC.

The mean total material balance ranged between 95.1% and 109.2%. The main radioactivity that

remained in the extracted soil ranged between 4.9% and 34.0% of AR. The cumulative volatile

radioactivity ranged between <0.1% to 55.4% of AR. The entire volatile radioactivity was 14

CO2. This

result indicates that all segments of the kasugamycin molecule were attacked.

Three minor metabolites were detected (max 8.1% of AR at day 30) by thin layer chromatography.

Attempt to characterize the metabolite that accounted for up to 8.1% revealed that the area should

contain several components. No further characterization was performed on these metabolites

because of their very low level.

The level of the parent substance decreased from a mean of 100.8% of AR at day 0 to 4.2% at the

end of the study, day 366. The half-life was calculated to be 40.8 days.

Characterization of the radioactivity associated with the soil extract or soil organic matter fractions

indicated that this radioactivity is metabolized by soil microorganisms and incorporated to soil organic

matter, especially with fulvic acid fraction.

In conclusion, the degradation rate of kasugamycin under aerobic conditions is moderate with a half-

life of 40.8 days. After 366 days of aerobic incubation, up to 55.4% of AR was detected as 14

CO2 and

30.6% of AR was in bound residues. A continuous decrease in the amount of organo-extractable

65


September 2013

radioactive material was observed. At the same time, a continuous increase in the amount of

radioactivity associated with high molecular weight materials or bound to soil fractions was observed,

accompanied by continuous evolution of carbon dioxide. This indicates that kasugamycin degraded

via biological catalysis, serving as a carbon source for soil microorganisms.

Conclusion: DT50 = 40.8 days

Flag: disregarded study

Test substance: kasugamycin monohydrochloride monohydrate

Endpoint: DT50

Value: 2 days and less than 1 day for the paddy soils and 1 days and less than 1 day for the upland

soils.

Reference: Degradation of kasugamycin in soil. Anonymous. 15/05/1991

Klimisch score: 3 no guideline followed, no GLP

Amendments/Deviations: not relevant, no guideline followed

GLP: no

Test guidelines: none

Dose levels: 1 ppm dry soil

Analytical measurements: not described

Study summary: a soil degradation study has been performed with 2 paddy soils and 2 upland soils.

The soil were incubated at 27oC, the aerobic/anaerobic conditions were not stated in the report. The

DT50 obtained are 2 days and less than 1 day for the paddy soils and 1 days and less than 1 day for

the upland soils.

Conclusion: The DT50 obtained are 2 days and less than 1 day for the paddy soils and 1 days

and less than 1 day for the upland soils. These results are considered as invalid.

Anaerobic Degradation in Soil (routes and rates) – Laboratory studies

Flag: supporting study: it is not an anaerobic degradation study but a degradation study in paddy

conditions (aerobic/anaerobic)



C)-

kasugamycin

Endpoint: DT50

Value: 90 days at 25oC

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September 2013

Reference: (14

-C)-kasugamycin: metabolic fate in soil under aerobic/anaerobic (paddy) conditions. CJ

Lewis Study number 1442/13. May 2003

Klimisch score: the study is valid but does not meet the requirements for an anaerobic degradation

study.


GLP: yes

Test guidelines: Japan MAFF test guidelines section 2-5-1 (2001)

Dose levels: 5 ppm

Analytical measurements: Liquid scintillation counting, HPLC

Study summary: The metabolism of kasugamycin has been studied in a flooded silt loam soil (pH 6.5;

2.8% of organic matter) under aerobic/anaerobic (paddy) conditions. The kasugamycin concentration

used was 5 ppm with respect to the surface water (1.2 ppm with respect to the total system).

Portions of sieved soil were added to glass vessels to a depth of 5 cm and flooded with reverse

osmosis water to a depth of 1.5 cm above the soil surface. Prior to dosing, they were maintained in

the dark at 25oC ± 2

oC for 21 days to allow the soil to become anaerobic. After dosing, they were in

the same conditions in an atmosphere of moistened air for 180 days. In addition to these non-sterile

samples, a set of sterile units were prepared in similar vessels but with bacterial air filters attached to

maintain sterility which was achieved by autoclaving.

At the end of the study (180 days), six non sterile control units were used to determine microbial

biomass and the sterility was checked in the sterile vessels.

Duplicate samples were taken from the non-sterile system for analysis at zero time and 3, 7, 13, 31,

59, 120 and 180 days. Duplicate samples were taken from the sterile system at 59 and 181 days.

Only low levels of applied radioactivity (< 1% for non-sterile units and < 5% for sterile units) were

present in surface water.

Radioactivity extracted from soil decreased from 92% of applied radioactivity (AR) to 43% (non-sterile

units) and 79% (sterile units) at the end of the study.

Non sterile units produced up to 26% carbon dioxide but no carbon dioxide was produced from sterile

units.

Bound residues fractionation at the end of the study gave the following results in non-sterile

conditions: 7.8% AR bound to fulvic acid; 2.9% to humic acid and 6.7% to humin. The corresponding

values for sterile conditions were: 9.4%, 1.0% and 3.2% respectively.

In non-sterile conditions, in addition to the main degradation product CO2, kasugamycinic acid was

detected (max 1.5% AR at 180 days), the unextracted residues accounted for 17.7% max at 59 days

and other components accounted for 4.3% max at the end of the study.

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September 2013

In the sterile system, kasugamycinic acid was also detected (10.1% AR at 181 days). The unextracted

residues accounted for 11.4% max at 181 days and other components accounted for 7.1% max at the

end of the study.

The data from the non-sterile units best fit a two-phase decay curve with a secondary slower rate of

degradation. The calculated DT50 was 90 days and the DT90 was 1052 days (extrapolation).

In sterile conditions the DT50 was 324 days (1st order) and the DT90 was 1075 days (extrapolation).

Conclusion: DT50 = 90 days at 25oC in paddy conditions

Soil photolysis

Flag: waiver

Test substance: not relevant

Endpoint: not relevant

Value: not relevant

Reference: Kasugamycin Waiver Request from Further Testing: Photodegradation on Soil

Klimisch score: not relevant

Amendments/Deviations: not relevant

GLP: no

Test guidelines: not relevant

Dose levels: not relevant

Analytical measurements: not relevant

Study summary: The applicant didn’t provide any study for the following reasons:

• The photodegradation in water study of kasugamycin shows that there was minimal photolytic

degradation of kasugamycin in pH 5 buffered solutions under continuous exposure to light at

wavelengths of 290-400 µm (the relevant wavelength range for photolysis).

• The UV-Visible absorption of kasugamycin technical study shows a lack of light absorption in the

relevant wavelength range of 290 - 400 µm, which corresponds to the relevant wavelengths that break

chemical bonds in pesticide molecules.

• Natural sunlight is nearly completely attenuated at the earth's surface at wavelengths <290 µm due

to atmospheric absorption.

• Comparative studies on pesticide photolysis in aqueous solution and on soil indicate that

kasugamycin photolysis through indirect mechanisms would be much less likely to occur on soil than

in aqueous solution.

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September 2013

Conclusion: The photodegradation of kasumycin in soil should not represent a major pathway

of degradation in the environment. No further study is necessary.

Adsorption/ desorption on soil particles

Flag: weight of evidence

Test substance: [D-Glucosamine-U-14

C]Kasugamycin. Also called [14

C]Kasugamycin

Endpoint: Adsorption and desorption coefficients (K) and Koc

Value: adsorption Koc from 6 to 324.

Reference: Soil Adsorption/Desorption of [14

C]Kasugamycin by the Batch Equilibrium Method. Mark

Schick. Study number 1820W. 11/08/2009

Klimisch score: 1


GLP: yes

Test guidelines: OECD 106 and US EPA 40 CFR 158, section series 163-1

Dose levels: five concentrations of approximately 3.0, 1.0, 0.3, 0.1, and 0.03 μg/mL

Study summary: The adsorption/desorption characteristics of [14

C]Kasugamycin were determined

using four soils which varied in texture, percentage of organic matter, pH and cation exchange

capacity. Preliminary trials demonstrated that a soil to solution ratio of 1:5 (4 g of soil and 20 mL of

0.01M CaCl2 solution) was optimal for North Dakota sandy clay loam and sandy loam soils, while a

soil to solution ratio of 1:10 (2 g of soil and 20 mL of 0.01M CaCl2) was optimal for a North Dakota

clay loam soil, and a soil to solution ratio of 3:1 (24 g of soil and 8 mL of 0.01M CaCl2) was optimal for

a Florida sand soil. Adsorption and desorption equilibration periods of 24 hours (North Dakota soils)

or 4 hours (Florida sand soil) were selected for the definitive study.

The definitive study was conducted at 25 °C in the dark with 4.0 g dry weight equivalent of North

Dakota sandy clay loam and sandy loam soils, 2.0 g dry weight equivalent of North Dakota clay loam

soil, and 24.0 g dry weight equivalent of Florida sand soil. The study was conducted with 20 mL

(North Dakota soils) or 8 mL (Florida sand soil) of 0.01 M aqueous calcium chloride, in 50 mL Teflon®

centrifuge tubes. The samples were dosed with [14

C]Kasugamycin dose solutions (500 μL for the

North Dakota soils and 200 μL for the Florida soil) prepared in 0.01M CaCl2 at five concentrations of

approximately 3.0, 1.0, 0.3, 0.1, and 0.03 μg/mL (ppm). After dosing, the tubes were reciprocally

shaken for 24 hours (North Dakota soils) or 4 hours (Florida soil) for each phase. For desorption, the

same volumes of fresh 0.01 M CaCl2 solution were added to the samples and the tubes were shaken

and processed as above.

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September 2013

The overall mass balance ranged from 93.4% to 106.1%. [14

C]Kasugamycin was stable in the

adsorption solutions, desorption solutions and control (no soil present) solution for the duration of the

equilibration period (greater than 94% Kasugamycin).

The Freundlich adsorption and desorption coefficients (K values) for [14

C]Kasugamycin were

calculated from concentrations in equilibrated solution (μg/mL) and soil (μg/g). Refer to table below.

Table 4: Adsorption/desorption of Kasugamycin

Soil type %

OC Kads Koc Kdes Koc

North Dakota

Sandy clay loam 3.4 4.54 134 8.20 241

North Dakota Clay

loam 3.3 10.68 324 14.50 439

North Dakota

Sandy loam 1.1 3.40 309 5.35 486

Florida Sand 0.5 0.03 6 No

desorption

Not

applicable

The adsorption K values were 4.54, 10.68, 3.40, and 0.03 for the sandy clay loam, clay loam, sandy

loam, and sand soils, respectively, with correlation coefficients r2 of 0.9997, 0.9999, 0.9999, and

0.9696, respectively. The KOC values for the North Dakota soils were 134 (sandy clay loam soil), 324

(clay loam soil), and 309 (sandy loam soil), indicating high to moderate mobility of [14

C]Kasugamycin

in these three soils. The KOC value for the Florida soil was 6, indicating very high mobility of

[14

C]Kasugamycin in the sand soil.

Desorption values were determined to be 8.20, 14.50, and 5.35 for the sandy clay loam, clay loam,

and sandy loam soils, respectively, with correlation coefficients of 0.9996, 1.0000, and 0.9995,

respectively. The KOC desorption values were 241, 439, and 486, respectively. Desorption was not

observed in the Florida sand soil used in the study. Radiocarbon recovered in desorption solutions

resulted from the entrained adsorption solution after the adsorption phase of the study. As a result,

Kdes and KOC values could not be accurately calculated.

Conclusion: Kads values were determined to be 4.54, 10.68, 3.40, and 0.03 for the sandy clay

loam, clay loam, sandy loam, and sand soils, respectively. The corresponding KOC values were

134, 324, 309, and 6, respectively; indicating that kasugamycin is very mobile to moderately

mobile in soils.

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September 2013

Lysimeter studies

Not provided. The available information regarding the mobility of kasugamycin in soil is considered as

sufficient.

Soil column leaching


Test substance: Kasugamycin and its metabolites: kasugamycinic acid and kasuganobiosamine

Endpoint: concentrations in eluate and different sections of the soil column

Value: Kasugamycin is immobile in soil, kasugamycinic acid is highly mobile.

Reference: Soil column leaching of Kasugamycin and related compounds in three soil types. Fujio

Ishijima. Study number HCRL9301. 20/10/1993

Klimisch score: 2 for kasugamycin and 3 for the metabolites.

Amendments/Deviations: The study does not fulfil the requirements of the current guidelines (OECD

312). Therefore the results are considered as partly valid only.

GLP: no

Test guidelines: US EPA Subdivision N, part 163-1

Dose levels: 1 mg of kasugamycin (corresponding to 5 kg/ha), 5 mg of kasugamycinic acid

(corresponding to 25 kg/ha) and 2 mg for kasuganobiosamine (corresponding to 10 kg/ha)

Analytical measurements: HPLC with fluorescence detector. The detection limits (LOD) were 10 µg

per soil segment for kasugamycin and kasuganobiosamine and 20 µg per soil segment for

kasugamycinic acid. In the eluate, they were 10 µg/L (kasugamycin and kasuganobiosamine ) and 20

µg/L (kasugamycinic acid). The LOD of kasugamycin was slightly higher than requested in the

guideline (0.5% of the initial dose, i.e. 5 µg per soil segment).

Study summary: The mobility of kasugamycin and of 2 of its degradation products was studied in a

leaching test on soil column. 3 soils were used for kasugamycin (light clay, sandy clay loam and loam

from paddy soils), only one (light clay for the 2 metabolites). This soil had the following properties (pH

6.8, 2.21 %OC and 32.2 % sand). This soil is the less mobility prone of the 3.

The recoveries were acceptable (between 70 and 110% of the initial dose) for kasugamycin and

kasuganobiosamine but analytical limitations (interferences) were observed with kasugamycinic acid

which had impacted the recoveries.

1 L of water was applied at the top of the column, equivalent to 508 mm rainfall. After the elution, the

soil in the column was segmented into 6 fractions at intervals of 5 cm.

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September 2013

All the kasugamycin residues were detected in the top segment (0-5 cm) of the columns with the 3

soils. The same pattern of distribution was obtained with kasuganobiosamine on the light clay soil.

Kasugamycinic acid was detected in all the six segments (10.0, 18.1, 30.2, 27.3, 10.3 and 4.1% in the

segments from the top to the bottom of the column). The residues of the 3 compounds in the eluates

were below the limit of detection in all columns.

Conclusion: Kasugamycin is immobile in soil, kasugamycinic acid is highly mobile. No

conclusion can be drawn concerning the mobility of kasuganobiosamine in soil due to

limitations in the protocol.

Field leaching studies

Not provided. Not necessary as the information regarding the mobility of kasugamycin in soil is

considered sufficient.

Aged residue column leaching

Not provided. Not necessary as the information regarding the mobility of kasugamycin and its

metabolite kasagamycinic acid in soil is considered sufficient.

Soil dissipation testing on a range of representative soils


Test substance: Kasugamycin and its metabolite, Kasugamycinic acid were analysed during the study

but the formulation Kasumin 2L was applied on soils

Endpoint: DT50 and mobility in soil

Value: mobility into layers below 15 cm is unlikely. No DT50 can be calculated from this study.

Reference: Kasugamycin Field Dissipation Study in Bare Ground. Janine E. Marin. Study number No.

1669W. 03/05/2007

Klimisch score: 2 valid with restriction as the analytical method is probably not appropriate.


GLP: yes

Test guidelines: Subdivision N, Chemistry: Environmental Fate, Section 164- 1, Terrestrial Field.

Dissipation Studies and NAFTA guidance document for conducting terrestrial field dissipation studies.

Dose levels: 4 applications of 21.8 g/acre (53.9 g a.i. /ha) with a 7-10 days interval.

Analytical measurements: The applied concentration was confirmed in the tank mixes and on filter

paper target pads.

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September 2013

Study summary: Field dissipation of the fungicide Kasugamycin and its metabolite, Kasugamycinic

acid, was measured in California, Washington, New York and Georgia soil following four applications

of the end-use product Kasumin 2L at an application rate 21.8 grams active ingredient per acre. The

four applications were applied with a 7-10 day application interval. This represents the maximum

proposed use pattern for Kasugamycin.

Tank mixes were analyzed for Kasugamycin concentration before and after each application. The

average percentage of nominal Kasugamycin in tank mixes from CA, WA, NY and GA are in the

range 88-116%. Kasugamycin application acceptability was also determined by the percent of

nominal recovered from target pads placed in the plots during the application. The average

Kasugamycin percent of nominal determined is in the range 79-109% for the 4 applications. These

results indicate that the applications were applied in a manner consistent with the desired application

rate and support the tank mix results.

Method validation was conducted for the analysis of Kasugamycin (KSM) and Kasugamycinic acid

(KSMA) in soil from all four sites. The validation for Kasugamycin and KSMA was conducted in

replicates at 0.01, 0.10 ppm, 1.0 ppm (for KSMA only) and 10 ppm (for KSM only). The limit of

quantitation (LOQ), determined as the lowest fortification yielding acceptable recoveries, was 0.01

ppm for Kasugamycin and Kasugamycinic acid. The concurrent recoveries with Kasugamycin and

Kasugamycinic acid carried out during sample analysis were nearly all within the acceptable range of

70-120% of nominal.

The average residue levels of Kasugamycin and Kasugamycinic acid in the 0-6” (0-15 cm) horizon

soil are presented in Table 5. Residues of Kasugamycin were found to decrease to <0.01 ppm within

14 days after last application at all sites, with one exception possibly resulting from a contaminated

sample. No residues of Kasugamycinic acid were detected in the 0- 6” (0-15 cm) soil horizon

throughout the analyses. No residues of Kasugamycin or Kasugamycinic acid were detected in the 6-

12” (15-30 cm) and 12-18” (30-45 cm) soil horizons at any time point at any of the four sites. The half-

life of Kasugamycin in soil was ~ 3 days in California and Washington and ~11 days in New York. A

definitive half-life could not be calculated for GA since no residues were detected at 3 days after last

application or later time points, but the results for the GA soil indicate the half-life is <3 days.

Table 5: Average residues (ppm) of kasugamycin (KSG) and its metabolite kasugamycinic acid (KSA) in the

0-15 cm segment

Time

(days)

New York Georgia California Washington

KSG KSA KSG KSA KSG KSA KSG KSA

-1 before 1st

application <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

1st

application 0.024 <0.01 0.018 <0.01 0.019 <0.01 0.018 <0.01

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September 2013

-1 before

2nd

application

0.022 <0.01 <0.01 <0.01 0.010 <0.01 <0.01 <0.01

2nd

application 0.042 <0.01 0.022 <0.01 0.026 <0.01 0.022 <0.01

-1 before 3rd

application 0.036 <0.01 <0.01 <0.01 0.019 <0.01 <0.01 <0.01

3rd

application 0.036 <0.01 0.020 <0.01 0.036 <0.01 0.027 <0.01

-1 before 4th

application 0.050 <0.01 <0.01 <0.01 <0.01 <0.01 0.016 <0.01

4th

application 0.057 <0.01 0.020 <0.01 0.020 <0.01 0.028 <0.01

3 after last

application 0.059 <0.01 <0.01 <0.01 0.020 <0.01 0.021 <0.01

7 after last

application 0.038 <0.01 <0.01 <0.01 <0.01 <0.01 0.010 <0.01

14 after last

application <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

1 month

after last

application

<0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

2 months

after last

application

<0.01 <0.01 ND* <0.01 ND* <0.01 ND* <0.01

4 months

after last

application

<0.01 <0.01 ND* <0.01 ND* <0.01 ND* <0.01

*ND: not determined

Conclusion: No appropriate DT50 was calculated from this study because values below the

LOQ occurred before the DT90 was reached. This indicates that the analytical method is

inadequate. However it can be concluded that the mobility kasugamycin into layers below 15

cm is unlikely.

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September 2013

General conclusion about environmental fate

The degradation rate of kasugamycin in aquatic environment is moderate in aerobic conditions (DT50

18-29 days), but very slow in anaerobic conditions (170 days). Hydrolysis in a limited extent and

photolysis can contribute to this degradation. Kasugamycinic acid is a significant metabolite (> 10% of

the applied dose) formed in all the aquatic degradations conditions tested.

The degradation study in soil in laboratory conditions shows that kasugamycin is persistent (DT50= 41

days). The degradation rate is slower in aerobic/anaerobic conditions (DT50 = 90 days). The only

major degradation product is CO2. Photolysis is not a significant pathway for the degradation of

kasugamycin in soils. The degradation studies in the field cannot be considered in the assessment

due to their deficiencies.

No bioaccumulation study was provided but on the basis of the log Kow value (< -1.96), it can be

considered that kasugamycin is not potentially bioaccumulative.

A weight of evidence approach has been applied to the studies related to the mobility in soils due to

the poor quality of the higher tier studies. Kasugamycin is considered to be immobile but its

metabolite kasugamycinic acid is highly mobile. A contamination of groundwater by kasugamycinic

acid is possible. The persistence of its metabolite in soil is unknown.

Significant metabolites:

Parent compound and kasugamycinic acid for the aquatic environment and sediments.

Parent compound only for soil (on the basis of the limited information provided).

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September 2013

Ecotoxicity - Robust study summaries for the active ingredient

Aquatic toxicity

Fish acute toxicity (Freshwater species)

Flag: Key study

Test substance: Kasugamycin Technical

Species: Rainbow trout (Oncorhynchus mykiss )

Type of exposure: 96 hr, static

Endpoint: LC50

Value: >120 mg a.i./L.

Reference: Kasugamycin Technical - Acute Toxicity to Rainbow Trout (Oncorhynchus mykiss) under static

conditions following OPPTS Draft Guideline 850.1075 AE Fournier. Study No. 13917.6114. 19/10/2009

Klimisch score: 1

Amendments/Deviations: none which had a negative impact on the results or interpretation of the study

GLP: yes

Test guidelines: OPPTS 850.1075 (draft, 1996)

No/group: 3 replicates of 10 fish/group

Dose levels: Limit test at 120 mg a.i./L (nominal and mean measured).

Analytical measurements: HPLC/UV

Study summary: The purpose of this study was to determine the acute toxicity of kasugamycin technical to a

representative freshwater fish species, the rainbow trout (Oncorhynchus mykiss), in a 96-hour static acute

toxicity test. Since <50% mortality was observed in the 120 mg a.i./L treatment level, the 96-hour LC50 was

empirically estimated to be >120 mg a.i./L. The No-Observed-Effect Concentration (NOEC), was 120 mg

a.i./L, based on a lack of mortality and sublethal effects at this concentration.

Conclusion: LC50 > 120 mg a.i./L

Flag: supporting study


Species: Fathead Minnow (Pimephales promelas)


Endpoint: LC50

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September 2013

Value: > 110 mg a.i./L

Reference: Kasugamycin Technical - Acute Toxicity to Fathead Minnow (Pimephales promelas) Under Static

Conditions, Following OPPTS Draft Guideline 850.1075. AE Fournier. Study No. 13917.6113. 21/10/2009

Klimisch score: 1

Amendments/Deviations: none which had a negative impact on the results or interpretation of the study

GLP: yes


No/group: 2 replicates of 10 per group

Dose levels: 7.5, 15, 30, 60 and 120 mg a.i./L (nominal); 7.4, 14, 28, 56 and 110 mg a.i./L (mean measured)



representative freshwater fish species, the fathead minnow (Pimephales promelas) in a 96-hour static toxicity

test. Since no concentration tested resulted in ≥ 50% mortality, the 96-hour LC50 was empirically estimated to

be > 110 mg a.i./L, the highest mean measured concentration tested. The No- Observed-Effect

Concentration (NOEC) was determined to be 110 mg a.i./L based on a lack of mortality and sublethal effects.


Fish acute toxicity (Marine species)



Species: Sheepshead Minnow (Cyprinodon variegatus).


Endpoint: LC50


Reference: Kasugamycin Technical - Acute Toxicity to Sheepshead Minnow (Cyprinodon variegatus). AE

Fournier. Study No. 13917.6112. 20/10/2009

Klimisch score: 1

Amendments/Deviations: none which had a negative impact on the results or interpretation of the study.

GLP: yes


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September 2013

No/group: 3 replicates of 10 fish/group

Dose levels: Limit test at 120 mg a.i./L (nominal); 110 mg a.i./L (measured)



representative saltwater fish species, the sheepshead minnow (Cyprinodon variegatus) in a 96-hour static

toxicity test. Since the concentration tested did not result in ≥ 50% mortality, the 96-hour LC50 was empirically

estimated to be > 110 mg a.i./L, the mean measured concentration tested. The No-Observed-Effect

Concentration (NOEC) was determined to be 110 mg a.i./L based on a lack of mortality and sublethal effects.


Invertebrates acute toxicity (Freshwater species)

Flag: Key study


Species: Daphnia magna


Endpoint: EC50

Value: > 90.7 mg a.i./L (mean measured)

Reference: Kasugamycin: Acute toxicity to Daphnia magna. Study number 1442/16. SD Mattock. August

2002

Klimisch score: 1

Amendments/Deviations: None that was considered to have affected the integrity and the outcome of the

study.

GLP: yes

Test guidelines: OECD 202 (1984); Japan MAFF 2-7-2-1; US EPA OPPTS draft guideline 850.1010

No/group: 2 replicates of 10 Daphnia/group

Dose levels: limit test at 100 mg/L (nominal)

Analytical measurements: HPLC-UV

Study summary: Daphnia magna was exposed for 48 hr to a nominal concentration of 100 mg/L of

Kasugamycin technical in a limit test in static conditions. The concentrations in the medium were measured

and the mean concentration is 90.7 mg/L. No immobilization has been observed at this concentration.

Therefore the EC50 is > 90.7 mg/L. The validity criteria defined in the guideline were met.

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September 2013

Conclusion: EC50 > 90.7 mg a.i./L

Invertebrates acute toxicity (Marine species)



Species: Mysids (Americamysis bahia)

Type of exposure: static, 96 hr

Endpoint: LC50


Reference: Kasugamycin Technical - Acute Toxicity to Mysids (Americamysis bahia), under static conditions,

following OPPTS Guideline 850.1035. AE Fournier. Study No. 13917.6115. 22/10/2009

Klimisch score: 1


GLP: yes


No/sex/group: 2 replicates of 10 per group



Study summary: The objective of this study was to estimate the acute toxicity (LC50) of kasugamycin

technical to mysids (Americamysis bahia, formerly Mysidopsis bahia) under static conditions. Since no

concentration tested resulted in ≥ 50% mortality, the 96-hour LC50 value was empirically estimated to be >

100 mg a.i./L, the highest mean measured concentration tested. The No-Observed-Effect Concentration

(NOEC) for this study was determined to be 100 mg a.i./L based on a lack of mortality and sublethal effects.




Species: Eastern Oyster (Crassostrea virginica)

Type of exposure: 96 hr, flow through

Endpoint: EC50

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September 2013


Reference: Kasugamycin Technical - Acute Toxicity to Eastern Oyster (Crassostrea virginica) Under Flow-

Through Conditions, Following OPPTS Guideline (Draft) 850.1025. DO York. Study No. 13917.6116.

30/11/2009

Klimisch score: 1

Amendments/Deviations: None that was considered to have affected the integrity and the outcome of the

study.

GLP: yes

Test guidelines: OPPTS (Draft) Guideline 850.1025 (1996)

No/group: 2 replicates of 20 per group

Dose levels: 3.8, 7.5, 15, 30, 60 and 120 mg a.i./L (nominal); 4.6, 8.7, 16, 33, 55 and 110 mg a.i./L (mean

measured)


Study summary: The objective of this study was to estimate the acute toxicity of kasugamycin technical to

Eastern oysters (Crassostrea virginica) under flow-through conditions. Since no concentration tested

resulted in ≥ 50% reduction in shell deposition, the 96-hour EC50 value was empirically estimated to be > 110

mg a.i./L, the highest mean measured concentration tested. The No-Observed-Effect Concentration (NOEC)

was determined to be 8.7 mg a.i./L, based on Williams’ Test.

Conclusion: EC50 > 110 mg a.i./L

Algae acute toxicity (Freshwater species)



Species: Pseudokirchneriella subcapitata

Type of exposure: static, 96 hours

Endpoint: ErC50

Value: 14 mg ai/L (95% confidence limits: 11-17)

Reference: Kasugamycin Technical – 96-Hour Acute Toxicity Test with Freshwater Green Alga,

Pseudokirchneriella subcapitata, Following OPPTS Draft Guideline 850.5400 and OECD Guideline 201.

Katherina A. Softcheck. Study No. 13917.6108 (19/11/2009)

Klimisch score: 1

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September 2013

Amendments/Deviations: no deviation that had an impact on the results or interpretation of the study

GLP: yes

Test guidelines: OPPTS Draft Guideline 850.5400 (1996) and OECD Guideline No. 201 (2006)

No/group: 6 replicates for the control; 3 per concentration

Dose levels: nominal concentrations: 0.20, 0.51, 1.3, 3.2, 8.0 and 20 mg a.i./L. Measured concentrations:

0.17, 0.44, 1.1, 2.8, 6.4 and 17 mg a.i./L

Analytical measurements: liquid chromatography/mass spectrometry (LC/MS/MS)

Study summary: The objective of this study was to determine the effect of kasugamycin technical on the

growth of the freshwater green alga, Pseudokirchneriella subcapitata. The results are based on mean

measured concentrations.

The mean measured exposure concentrations ranged from 80 to 87% of nominal concentrations and defined

the treatment levels tested as 0.17, 0.44, 1.1, 2.8, 6.4 and 17 mg a.i./L.

The validity criteria defined by the OECD guideline are shown in the following table. The study is considered

to be valid.

Table 6: Validity criteria

Acceptance Criteria for Control Study Results

16 x 104 cells/mL at 96 hours 140.25 x 10

4 cells/mL

Mean daily growth rate CV < 35% 22%

0 - 96-hour growth rate CV < 10% 4.1%

To determine the algistatic/algicidal properties of the test substance, an aliquot was removed at test

termination from a composite of the replicates of the 20 mg a.i./L nominal test solution. The estimated cell

density of the 100-mL solution prepared from the 20 mg a.i./L test solution was 0.1067 x 104 cells/mL. The

subculture was maintained in the same conditions as in the main study for six days. After four days, a cell

density of 345 x 104 cells/mL was observed in the subculture. This observation indicates that kasugamycin

technical has an algistatic, rather than algicidal effect on the growth of Pseudokirchneriella subcapitata at a

nominal concentration of 20 mg a.i./L.

Table 7: Biological results

Biological

parameter

Based on Mean Measured concentrations (mg a.i./L)

EC50 (95% confidence limits) NOEC

96-hour cell 4.2 (3.0-5.0) 1.1

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September 2013

density

EyC10 (95%

confidence

limits)

EyC20 (95%

confidence

limits)

EyC50 (95%

confidence

limits)

NOEC

72-hour yield 0.55 (0.17-1.5) 0.84 (0.20-1.9) 3.3 (2.3-4.2) 1.1

ErC10 (95%

confidence

limits)

ErC20 (95%

confidence

limits)

ErC50 (95%

confidence

limits)

NOEC

72-hour average

growth rate 2.2 (0.41-3.0) 4.4 (3.6-5.1) 14 (11-17) 6.4

a

a The NOEC was determined to be 6.4 mg a.i./L based on Kruskal-Wallis' test. Since a 29% inhibition was observed at

this treatment level, a more conservative estimate is the ErC10 (i.e., 2.2 mg a.i./L), as suggested by the OECD 201 Guideline.

Conclusion: ErC50 = 14 mg a.i./L. Kasugamycin effect is algistatic at 20 mg a.i./L rather than algicidal.



Species: Navicula pelliculosa

Type of exposure: 96 hr static

Endpoint: ErC50

Value: > 100 mg/L

Reference: Kasugamycin Technical – 96-Hour Toxicity Test with the Freshwater Diatom, Navicula

pelliculosa, following OPPTS Draft Guideline 850.5400 and OECD Guideline 201. Katherina A. Softcheck.

Study No. 13917.6109. 30/10/2009

Klimisch score: 1

Amendments/Deviations: none that had a negative impact on the results or interpretation of the study

GLP: yes





Study summary: The purpose of this study was to determine the effect of kasugamycin technical on the

growth of the freshwater diatom, Navicula pelliculosa. The results are based on mean measured

concentrations of kasugamycin.

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September 2013


the treatment levels tested as 7.0, 13, 27, 52 and 110 mg a.i./L.

The validity criteria defined by the OECD guideline are shown in the following table. The study is considered

to be valid.




4 cells/mL

Mean daily growth rate CV < 35% 27.79%


Table 9: Biological results

Biological

parameter



96-hour cell

density 90 (81-97) 52

EyC10 (95%

confidence

limits)

EyC20 (95%

confidence

limits)

EyC50 (95%

confidence

limits)

NOEC

96-hour yield 46 (37-60) 57 (45-68) 90 (81-97) 52

ErC10 (95%

confidence

limits)

ErC20 (95%

confidence

limits)

ErC50 (95%

confidence

limits)

NOEC

96-hour average

growth rate 76 (68-84) 104 (96-110) >100 (NA) a 52

a NA = Not Applicable. EC value was empirically estimated; therefore, 95% confidence limits could not be calculated

Conclusion: ErC50 >100 mg a.i./L

Flag: Key study


Species: Anabaena flos-aquae

Type of exposure: static, 96 hours

Endpoint: ErC50

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September 2013

Value: 1.3 mg ai/L (95% confidence limits: 1.0-2.0)

Reference: Kasugamycin Technical – 96-Hour Toxicity Test with the Freshwater Blue-Green Alga, Anabaena

flos-aquae, Following OPPTS Draft Guideline 850.5400 and OECD Guideline No. 201. K.A. Softcheck. Study

No. 13917.6111

Klimisch score: 1


GLP: yes



Dose levels: nominal concentrations: 0.10, 0.26, 0.64, 1.6, 4.0 and 10 mg a.i./L. Mean measured

concentrations: 0.080, 0.22, 0.51, 1.3, 3.3 and 9.0 mg a.i./L



growth of the freshwater blue-green alga, Anabaena flos-aquae. The results are based on mean measured

concentrations.


the treatment levels tested as 0.080, 0.22, 0.51, 1.3, 3.3 and 9.0 mg a.i./L.

The validity criteria defined by the OECD guideline are shown in the following table. The mean daily CV for

growth rates was 79%. Although the criterion suggests that this value should not exceed 35%, growth of A.

flos-aquae is typically more variable and more difficult to count accurately due to its filamentous structure, in

contrast to unicellular green algae, which was used as the basis for this OECD criterion. Despite that, the

study is considered to be valid.




4 cells/mL



To determine the algistatic/algicidal properties of the test substance, an aliquot was removed at test

termination from the composite of the 10 mg a.i./L nominal solution. The estimated cell density of the 100-mL

solution was 0.0033 x 104 cells/mL. The subculture was maintained in the same conditions as in the main

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September 2013

study for six days. After six days, a cell density of 7.25 x 104 cells/mL was observed in the subculture. This

observation indicates that the test substance has an algistatic, rather than algicidal effect on the growth of

Anabaena flos-aquae at 10 mg a.i./L.

Table 3 Biological results

Biological

parameter



96-hour cell

density 0.76 (0.57-0.92) 0.22

EyC10 (95%

confidence

limits)

EyC20 (95%

confidence

limits)

EyC50 (95%

confidence

limits)

NOEC

96-hour yield 0.11a 0.17

a 0.76 (0.57-0.91) 3.3

b

ErC10 (95%

confidence

limits)

ErC20 (95%

confidence

limits)

ErC50 (95%

confidence

limits)

NOEC

96-hour average

growth rate 0.53 (0.33-0.63) 0.70 (0.61-0.83) 1.3 (1.0-2.0) 0.51

a Determined by linear regression. Corresponding 95% confidence limits could not be determined.

b Based on Kruskal-Wallis’ Test, the 96-Hour NOEC was determined to be 3.3 mg a.i./L. Since 97% reduction of yield

was observed at this treatment level, a more conservative estimate of the NOEC value is the EyC05,EyC10 or EyC20 value (0.092, 0.11 or 0.17 mg a.i./L, respectively, determined by linear regression) as suggested by the OECD guideline.

Conclusion: ErC50 = 1.3 mg a.i./L. Kasugamycin effect is algistatic at 10 mg a.i./L rather than algicidal.

Algae acute toxicity (Marine species)



Species: Skeletonema costatum


Endpoint: ErC50


Reference: Kasugamycin Technical - 96-Hour Toxicity Test with the Marine Diatom, Skeletonema costatum,

following OPPTS Draft Guideline 850.5400 and OECD Guideline No. 201. Katherina A. Softcheck. Study No.

13917.6110. 04/11/2009

Klimisch score: 1


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September 2013

GLP: yes

Test guidelines: Draft Test Guidelines 850.5400 (1996); OECD 201 (2006)

No/group: 3 replicates per treatment group and 6 for the control




growth of the marine diatom, Skeletonema costatum. The results are based on mean measured

concentrations of kasugamycin.


the treatment levels tested as 6.0, 12, 23, 47 and 94 mg a.i./L.

The validity criteria defined by the OECD guideline are shown in the following table. The mean daily CV for

growth rates was 52%. Although the criterion suggests that this value should not exceed 35%, growth of S.

costatum is typically more variable and more difficult to count accurately due to the fact that it grows in short

chains and groups, in contrast to unicellular green algae, which were used to define this OECD criterion.

Despite that, the study is considered to be valid.




4 cells/mL at 96 hours



Table 5 Biological results

Biological

parameter



96-hour cell

density > 94 (NA) a 94

EyC10 (95%

confidence

limits)

EyC20 (95%

confidence

limits)

EyC50 (95%

confidence

limits)

NOEC

96-hour yield > 94 (NA) a > 94 (NA)

a > 94 (NA) a 94

ErC10 (95%

confidence

ErC20 (95%

confidence

ErC50 (95%

confidence NOEC

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September 2013

limits) limits) limits)

96-hour average

growth rate > 94 (NA) > 94 (NA)

a > 94 (NA) a 94

a NA = Not Applicable. EC value was empirically estimated; therefore, 95% confidence limits could not be calculated

Conclusion: ErC50 > 94 mg a.i./L

Aquatic plants acute toxicity (Freshwater species)

Flag: key study


Species: Lemna gibba

Type of exposure: 7 days. Renewal on day 4.

Endpoint: EC50


Reference: Kasugamycin Technical - 7-Day Toxicity Test with Duckweed (Lemna gibba) following OPPTS

Draft Guideline 850.4400 and OECD Guideline No. 221. Katherina A. Softcheck. Study No. 13917.6107.

02/11/2009

Klimisch score: 1


GLP: yes

Test guidelines: OECD 221 (2006) and draft OPPTS 850.4400 (1996).

No/group: 3 replicates of 12 fronds for the test concentrations and 6 replicates of 12 fronds for the control.

Dose levels: 7.5, 15, 30, 60 and 120 mg a.i./L (nominal), 6.1, 11, 26, 63 and 110 mg a.i./L (mean measured)

Analytical measurements: liquid chromatography with mass spectrometry (LC/MS/MS)

Study summary: The purpose of this study was to determine the effects of kasugamycin technical on the

growth of the duckweed, Lemna gibba. The results are based on mean measured concentrations of

kasugamycin.

The guidelines require that the doubling time of frond number in the control must be less than 2.5 days,

which corresponds to a seven-fold increase in seven days. During this study, the doubling time was 1.5

days, which meets the validity criterion of the guidelines.

The yield EC50 for frond density was 85 mg a.i./L (95% confidence limits: 64-94). The ErC50 was > 110 mg

a.i./L for both frond density and dry weight).

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September 2013

Conclusion: ErC50 > 110 mg a.i./L (for frond density and dry weight).

Fish chronic toxicity (Freshwater species)

Flag: Key study


Species: Pimephales promelas

Type of exposure: flow-through, 32 days, ELS

Endpoint: NOEC

Value: 9.5 mg a.i./L

Reference: Kasugamycin Technical – Early Life-Stage Toxicity Test with Fathead Minnow (Pimephales

promelas), following OECD Guideline #210 and OPPTS Draft Guideline 850.1400. Study number

13917.6117. 22/10/2009

Klimisch score: 1

Amendments/Deviations: deviation in the study conditions (temperature and specific conductance of the

water) which was considered as negligible as the survival in the control met the validity criteria.

GLP: yes

Test guidelines: OECD 210 and OPPTS Draft Guideline 850.1400

No/group: 2 replicates of 10 fry

Dose levels: nominal: 0.63, 1.3, 2.5, 5.0 and 10 mg a.i./L; mean measured concentration: 0.60, 1.1, 2.1, 4.8

and 9.5 mg a.i./L

Analytical measurements: liquid chromatography with mass spectrometry (LC/MS/MS)

Study summary: During the study, daily measurements of the test solutions established a dissolved oxygen

concentration range of 7.0 to 9.1 mg/L. Continuous temperature monitoring of the control demonstrated that

the temperature ranged from 24 to 26 ºC throughout the exposure. Measurements of pH ranged from 6.9 to

7.4. Weekly characterization of the high, low and control solutions established that the water quality

parameters measured were not affected by the established concentration gradient of kasugamycin technical

and were maintained throughout the 32-day exposure within an acceptable range for the promotion of

fathead minnow embryo hatchability, larval survival, and growth.

Mean measured concentrations ranged from 82 to 96% of the nominal levels and defined the treatment

levels tested as 0.60, 1.1, 2.1, 4.8 and 9.5 mg a.i./L.

Due to the mortalities associated with the fungal infection in one replicate of the control, hatching success

was calculated and statistically analyzed two ways. These data were evaluated both including and excluding

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these mortalities. At the completion of hatch (day 4), hatching success in the control averaged 72%,

including mortalities due to fungal infection; hatching success in the control averaged 77% if these mortalities

are excluded. Embryo hatching success in the 0.60, 1.1, 2.1, 4.8 and 9.5 mg a.i./L treatment levels averaged

89, 83, 81, 88 and 80%, respectively. There was no significant difference in embryo hatching success among

organisms exposed to any of the treatment levels tested compared to the hatching success of the control

organisms. The results of these analyses do not differ whether the mortalities due to the fungal infection are

included or excluded from the calculation of hatching success. At the completion of the hatching period (day

4), the percent of normal larvae in the control and all treatment levels tested was 100%. Since percent of

normal larvae was 100% in all treatment levels tested and no deformed larvae were observed at hatch, this

endpoint was not statistically analyzed. No delays in hatching were observed in any treatment group

compared to the control. Following 28-days post-hatch exposure (day 32), larval survival in the control group

averaged 83%. Larval survival in the 0.60, 1.1, 2.1, 4.8 and 9.5 mg a.i./L treatment levels averaged 85, 98,

88, 93 and 90%, respectively. There was no significant difference in larval survival in total larvae length, dry

weight among organisms exposed to any of the treatment levels tested compared to the control organisms.

Based on embryo hatching success, larval survival and larval growth (length and dry weight), the No-

Observed- Effect Concentration (NOEC) was determined to be 9.5 mg a.i./L, the highest mean measured

concentration tested. The Lowest-Observed-Effect Concentration (LOEC) for kasugamycin technical and

fathead minnow was determined to be > 9.5 mg a.i./L.

Conclusion: NOEC = 9.5 mg a.i./L

Fish chronic toxicity (Marine species)

Not provided: not required , the information on freshwater organisms is appropriate for risk assessment and

classification purposes.

Invertebrates chronic toxicity (Freshwater species)

Flag: key study



Type of exposure: semi-static, 3 renewals per week

Endpoint: NOEC

Value: 50 mg a.i./L for reproduction, growth and parent mortality

Reference: The chronic toxicity (21 days) of kasugamycin to Daphnia magna. AHC Groeneveld et al. Study

number CDNL 51.006 (18/05/1992)

Klimisch score: 1

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September 2013


GLP: yes

Test guidelines: US EPA 72-2 (1982), OECD 202 part 2 (1984)

No/group: 4 replicates of 10

Dose levels: 0, 6, 12, 25, 25, 50, 100 mg a.i./L. Measured concentration are between 100 and 106% of the

nominal.

Analytical measurements: HPLC on new and old culture medium

Study summary: The purpose of this study was to estimate the 21-day chronic toxicity and effects on

reproduction of kasugamycin technical to Daphnia magna at nominal test concentrations of 0, 6, 12, 25, 25,

50, 100 mg a.i./L under semi-static conditions (3 renewals/week).

Samples were taken from fresh and old medium and analysed by HPLC. The concentration of kasugamycin

remained stable during the test (100 to 106% of the nominal were determined).

The validity criteria of the guideline were met except the pH which had to remain in a range ± 0.3 (the

observed pH varied between 7.2 and 7.9) however as the mortality in the control was < 20% at the end of the

test (12.5%) and the mean number of young / female was > 40 (50), this pH change was considered

acceptable.

The NOEC for parental mortality, growth (length) and reproduction was 50 mg a.i./L.

Conclusion: The NOEC for parental mortality, growth (length) and reproduction was 50 mg a.i./L.

Invertebrates chronic toxicity (Marine species)

Not provided: not required, the information on freshwater organisms is appropriate for risk assessment and

classification purposes.

General conclusion about aquatic ecotoxicity

Kasugamycin shows a low acute and chronic toxicity on fish and aquatic invertebrates. However, its toxicity

for algae varies depending on species. On the basis of the most sensitive species, Kasugamycin is classified

9.1B (toxic for the aquatic organisms).

No ecotoxicity study has been provided with the significant metabolite kasugamycinic acid. The applicant

explained that kasugamycin will not contaminate water bodies because it is not mobile in soils. Kasugamycin

is able to contaminate water bodies via spray drift, and kasugamycin will be broken down in this metabolite in

water so this is a data gap.

8.2.2. Sediment toxicity

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September 2013

Not provided: this is a data gap as around 40% of kasugamycin is distributed in the sediments according to

the results of the water/sediment study. However the results of the chronic study on aquatic invertebrates

(Daphnia) do not show any effects on reproduction, it is acceptable to waive this study.

8.2.3. Soil toxicity

Soil macro-invertebrates acute toxicity

Flag: key study


Species: Eisenia fetida

Type of exposure: static, 14 days

Endpoint: LC50

Value: > 1000 mg a.i./kg dry soil

Reference: Kasugamycin technical: a 14-day acute toxicity test with the earthworm Eisenia fetida. K.

Nienstedt. Study number 1081.001.630. 29/08/2005

Klimisch score: 1

Amendments/Deviations: some deviations from the study plans which were not considered to have impacted

the results of the study.

GLP: yes

Test guidelines: OECD 207 (1984)

No/sex/group: 4 replicates of 10 earthworms per treatment

Dose levels: 62.5; 125; 250; 500 and 1000 mg a.i./kg dry soil

Analytical measurements: not required

Study summary: Groups of 40 Eisenia fetida were exposed for 14 days to Kasugamycin technical at

concentrations from 62.5 to 1000 mg a.i./kg, on artificial soil. On day 7 and 14, mortality and health

assessments were performed. Individual worms were weighed on days 0 and 14. Repellent effects were

evaluated on days 0 and 7.

On days 7 and 14, no mortality was observed in any of the treatments. Therefore, the LC50 was estimated to

be higher than 1000 mg a.i /kg dry soil. Sub-lethal effects were observed at 250 mg a.i /kg dry soil: 2.5% of

the worms were lethargic. In the 500 and 1000 mg a.i /kg dry soil treatments, the worms showed less activity

than in the lower test item concentrations and the control treatment.

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September 2013

The weight loss from day 0 to day 14 was not statistically significant and not dose responsive. The NOEC for

body weight is 1000 mg a.i /kg dry soil.

Conclusion: LC50 > 1000 mg a.i./kg dry soil

Soil macro-invertebrates chronic toxicity

Not provided: the chronic toxicity of kasugamycin on earthworms cannot be predicted from the acute test.

Kasugamycin is persistent in soil (DT50 > 30 days) so this is a data gap.

Non-target plants acute toxicity

Not provided: no study performed with the active ingredient but the formulation Kasumin 2L has been tested.

Nitrogen transformation test

Flag: key study

Test substance: Kasugamycin

Species: soil microorganisms

Type of exposure: static for 8 weeks

Endpoint: Effect or not based on the comparison of t1/2 values of the ammonia concentrations

Value: No effect was observed at 2.5 kg ai/ha. For both soils, an inhibition of the ammonium oxidation was

observed at kasugamycin concentrations of 5 to 25 kg ai/ha.

Reference: The effects of Kasugamycin on nitrification in soil. Melkebeke. Report number 231795.

14/09/1998

Klimisch score: 2, the guideline followed is different from the OECD guideline 216: ammonium added to the

soil instead of powdered plant meal; soil characteristics are different; endpoint is different. However it is a

validated guideline and the study was performed according to GLP, and the results of this study are fully

relevant for the purpose of the evaluation of the effects of the test substance on the N cycle in soil.

Amendments/Deviations: none compared to the guideline

GLP: yes

Test guidelines: Nederlandse Norm NEN 5795. Determination of the influence of chemical substances on

nitrification processes in soil (1988)

Dose levels: 2.5 – 5 – 12.5 - 25 kg ai/ha

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Study summary: The effects of Kasugamycin on the nitrification process was studied in two different soil

types being Speyer soil 2.2 (loamy sand) and Cranfield soil No. 164 (silt loam). Both soils were amended

with an aqueous mixture of a nitrogen source and the test substance. Ammonium nitrogen was added as an

aqueous solution of (NH4)2SO4 equivalent to 10 mg nitrogen. The test concentration levels were 2.5 – 5 –

12.5 - 25 kg ai/ha, equivalent to 3.33 – 6.67 – 16.7 – 33.3 mg a.i./kg dry weight soil. The test mixture (5 mL)

was added to 100 g (dry weight) equilibrated soil samples. After treatment the samples were incubated at

20°C ± 2°C and at 60% of their maximum water capacity over a period of 8 weeks.

At various time intervals (0, 1, 2, 4, 6 and 8 weeks) ammonia, nitrite and nitrate levels were measured. The

t1/2 values of the ammonia concentrations were calculated and compared with the untreated soil.

Table 6 Calculated T1/2 values of the NH4-N concentration (rounded values)

Applic.

Rate

kg ai/ha

Speyer 2.2 Cranfield 164

T1/2

(days)

Ratio

(treated/

untreated)

Difference

(treated-

untreated) Effect

T1/2

(days)

Ratio

(treated/

untreated)

Difference

(treated-

untreated) Effect

Control 17.91 8

2

2.5 18.01 1.0 0.1 No effect 7.4

3 0.9 0.6 No effect

5 24.01 1.3 6.1 Effect 9.5

2 1.2 1.5 Effect

12.5 27.91 1.6 9.9 Effect 10.7

3 1.3 2.7 Effect

25 24.11 1.3 6.1 Effect 15

2 1.9 7 Effect

1 T1/2 determined using linear regression with 4 or 5 time points

2 T1/2 read from the curve

3 T1/2 determined using first order model

According to the guideline, treatments with t1/2 values showing a ratio > 1.15 and a difference > 1 day with

the control treatment are evaluated to have a restraining influence on the nitrification process in soil.

No effect was observed at 2.5 kg ai/ha. For both soils, an inhibition of the ammonium oxidation was

observed at kasugamycin concentrations of of 5, 12.5 and 25 kg ai/ha.

Conclusion: No effect was observed at 2.5 kg ai/ha (application rate in New Zealand is 0.1 kg a.i./ha) .

For both soils, an inhibition of the ammonium oxidation was observed at kasugamycin

concentrations of 5 to 25 kg ai/ha.

Carbon transformation test

Not provided: This study is required because kasugamycin exhibits bactericidal activity.

General conclusion about soil ecotoxicity

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September 2013

The acute toxicity of kasugamycin on earthworm is low. The effects on soil micro-organisms (N

transformation) occur at concentration far above the application rate recommended in New Zealand (100 g

ai/ha).There are gaps in the data package provided by the applicant regarding the chronic effects on

earthworms and the carbon transformation in soil. So the full assessment of the soil ecotoxicity is not

possible.

Kasugamycin is not classified as 9.2 on the basis of the available information.

Terrestrial vertebrate toxicity

For effects on terrestrial vertebrates other than birds, refer to the mammalian toxicity section.

Oral toxicity

Flag: Key study


Species: Mallard Duck (Anas platychynchos)

Type of exposure: acute, 14 days of observation.

Endpoint: LD50

Value: > 2000 mg a.i./kg bw

Reference: Kasugamycin Hydrochloride: Acute Oral Toxicity Test (LD50) with the Mallard Duck (Anas

platychynchos). JM. Stafford. Study No. 13862.4102. 28/06/2006

Klimisch score: 1

Amendments/Deviations: no deviation that had a negative impact on the results or interpretation of the study.

GLP: yes

Test guidelines: OPPTS Guideline 850.2100, Japan MAFF Guideline 2-8-4-1

No/sex/group: 5

Dose levels: Limit test at 2000 mg a.i./kg bw


Study summary: Kasugamycin was administered to groups of 10 Mallard ducks at a dose of 2000 mg/kg bw.

The dosing was followed by a 14-day observation period.

No mortality, morbidity, or clinical signs of intoxication were observed during this study in any individual

tested.

No significant differences in mean feed consumption per bird per day among the treatment group tested and

the control group at any interval.

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September 2013

No significant differences in proportional body weight change, calculated at day 0 – 7 or day 0 - 14 among

males or females, were observed when comparing the treatment and control group.

Since no concentration tested resulted in mortality and no adverse effects were observed, the LD50 was

empirically estimated to be > 2000 mg a.i./kg body weight, the highest nominal concentration tested. The No-

Observed-Effect Level (NOEL) for mortality was determined to be 2000 mg a.i./kg body weight.

Conclusion: LD50 > 2000 mg a.i./kg bw

Flag: Key study


Species: Bobwhite quail (Colinus virginianus)

Type of exposure: acute, 14 days of observation

Endpoint: LD50


Reference: Kasugamycin Hydrochloride: Acute Oral Toxicity Test (LD50) with Northern Bobwhite Quail

(Colinus virginianus). Study No. 13862.4103. JM. Stafford 10/10/2006

Klimisch score: 1

Amendments/Deviations: none that was considered to have affected the integrity and the outcome of the

study.

GLP: yes

Test guidelines: OPPTS Guideline 850.2100; Japan MAFF Guideline 2-8-4-1

No/sex/group: 5

Dose levels: limit test at 2000 mg a.i./kg bw


Study summary: Kasugamycin was administered to groups of 10 Bobwhite quails at a dose of 2000 mg/kg

bw. The dosing was followed by a 14-day observation period.

No mortality or morbidity, were observed during this study in any individual tested.

Diarrhea was observed among all treatment animals through day 2 post treatment.

No significant differences in mean feed consumption per bird per day among any of the treatment groups

tested and the control group at any interval.

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September 2013

Significant differences in proportional body weight change, calculated at day 0 – 7 and day 0 - 14 were

observed when comparing the treatment and control group. This may be related to diarrhea. The birds were

in the process of regaining their weight back during week 2.

Since no concentration tested resulted in mortality, the LD50 was empirically estimated to be > 2000 mg

a.i./kg body weight, the highest nominal concentration tested. The No-Observed-Effect Level (NOEL) for

mortality was determined to be 2000 mg a.i./kg body weight.

Conclusion: LD50 > 2000 mg a.i./kg body weight

Flag: Key study


Species: Zebra Finch (Taeniopygia guttata).

Type of exposure: acute, 14 days of observation

Endpoint: LD50


Reference: Kasugamycin Technical – Acute Oral Toxicity Test (LD50) with Zebra Finch (Taeniopygia guttata).

CA. Redmond. Study No. 13917.4102. 09/09/2009

Klimisch score: 1

Amendments/Deviations: none.

GLP: yes

Test guidelines: OPPTS Guideline 850.2100 (1996)

No/sex/group: 5

Dose levels: limit test at 2000 mg a.i./kg bw


Study summary: The objective of this test was to determine whether the LD50 of kasugamycin technical orally

administered to zebra finches is greater than 2000 mg a.i./kg body weight. This study was conducted as a

14-day limit test at a nominal concentration of 2000 mg a.i./kg body weight.

No mortality was observed among birds exposed in the 2000 mg a.i./kg body weight group or the control

group during the test. No regurgitation of the dose was observed. Four treatment birds displayed slight

piloerection during the first two hours post-dosing but were normal thereafter.

No significant differences in mean feed consumption per bird per day among any of the treatment groups

tested and the control group at any interval.

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September 2013

Since no concentration tested resulted in mortality, the LD50 was empirically estimated to be > 2000 mg

a.i./kg body weight, the highest nominal concentration tested. The No-Observed-Effect Level (NOEL) for

mortality was determined to be 2000 mg a.i./kg body weight.

Conclusion: LD50 > 2000 mg a.i./kg body weight

Dietary toxicity

Flag: Key study


Species: Bobwhite Quail (Colinus virginianus)

Type of exposure: in diet for 5 days

Endpoint: LC50

Value: > 5000 mg a.i./kg feed

Reference: Kasugamycin Hydrochloride: Dietary Toxicity Test (LC50) with Northern Bobwhite Quail (Colinus

virginianus). Study No. 13862.4101. JM. Stafford. 27/11/2006

Klimisch score: 1

Amendments/Deviations: no deviations that have a negative impact on the results or interpretation of the

study

GLP: yes

Test guidelines: OPPTS 850.2200 (1996); OECD 205 (1984); Japan MAFF Guideline 2-8-4-2

No/sex/group: 12 birds per cage for treatment groups, 24 birds for the control

Dose levels: 675, 1113, 1836, 3030 and 5000 mg a.i./kg feed (nominal); 581, 1079, 2058, 2882 and 4858 mg

a.i./kg feed (mean measured concentrations)


Study summary: No mortality was observed among quails exposed to the treatment levels tested (675, 1113,

1836, 3030 and 5000 mg a.i./kg) or the control group. The average daily exposure was: 130.91, 214.16,

367.20, 640.96 and 977.92 mg a.i./kg body weight/day.

Proportional body weight gain in the 675 mg a.i./kg feed group was significantly greater than the control

during the day 0 to 5 (treatment) and day 0 to 8 (overall) interval. Proportional body weight gain in the 675

and 1113 mg a.i./kg feed groups was significantly less than the control during the day 5 to 8 (post-treatment)

interval.

LC50 > 5000 mg a.i./kg feed

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September 2013

NOEC (mortality and weight change) = 5000 mg a.i./kg feed

Conclusion: LC50 > 5000 mg a.i./kg feed (> 977.92 mg a.i./kg body weight/day)

Flag: Key study


Species: Mallard Duck (Anas platyrhynchos).

Type of exposure: in diet for 5 days

Endpoint: LC50

Value: > 5000 mg a.i./kg feed

Reference: Kasugamycin Hydrochloride: Dietary Toxicity Test (LC50) with the Mallard Duck (Anas

platyrhynchos). JM Stafford. Study number 13862.4100. 27/11/2006

Klimisch score: 1

Amendments/Deviations: no deviations that have a negative impact on the results or interpretation of the

study

GLP: yes

Test guidelines: OPPTS 850.2200 (1996); OECD 205 (1984); Japan MAFF Guideline 2-8-4-2

No/sex/group: 12 birds per cage for treatment groups, 24 birds for the control

Dose levels: 675, 1113, 1836, 3030 and 5000 mg a.i./kg feed (nominal); 581, 1079, 2058, 2882 and 4858 mg

a.i./kg feed (mean measured concentrations)


Study summary: No mortality was observed among ducks exposed to the treatment levels tested (675, 1113,

1836, 3030 and 5000 mg a.i./kg) or the control group. The average daily exposure was: 142.97; 255.16;

400.61; 654.43 and 1118.95 mg a.i./kg body weight/day.

ANOVA of measured body weights detected a statistically significant mean difference between the control

group and the 3030 mg a.i./kg feed treatment group on day 5 of the test, with the treatment group mean

weight being lower than the control group mean. No other significant differences were detected in mean

measured body weights among groups on days 0, 5 and 8.

All test groups gained weight steadily throughout the study. However, statistically significant lower

proportional weight gain was observed in the 675 mg a.i./kg feed group , 3030 mg a.i./kg feed group, and

5000 mg a.i./kg feed group during the days 0 to 5. The 675 mg a.i./kg feed group and 5000 mg a.i./kg feed

group also gained proportionally less weight than the control group during the day 0 to day 8 interval.

However, proportional changes in body weight among the 1113 mg a.i./kg feed group, 1836 mg a.i./kg feed

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September 2013

group, and 3030 mg a.i./kg feed group were significantly greater than that of the control group during the day

5 to day 8 interval.

While the 675 mg a.i./kg feed group demonstrated statistically lower proportional weight gain than the

controls during two study intervals, no such differences were detected during any study intervals in the next

two higher exposure levels (1113 and 1836 mg a.i./kg feed). This suggests the reduced proportional weight

gain in the 675 mg a.i./kg feed group was likely an anomaly related to the dynamics of dominance

differences at the feed tray among individual birds in the group or inherent growth characteristics of certain

individual birds rather the influence of the test substance. This assessment is supported by the pattern of

proportional growth during the day 5 to day 8 interval, during which all groups were placed on clean feed. In

the absence of the test substance in the diet, the 1113, 1836 and 3030 mg a.i./kg feed groups gained a

significantly greater percentage of body weight than the control group, indicating that the day 0 to day 5

reduction in proportional weight gain in these groups was the result of the test substance. The same pattern

of recovery in proportional weight gain was not observed in the 675 mg a.i./kg feed group during the day 5 to

day 8 interval, indicating that the lower proportional weight gain observed in this group was the result of

something other than the test substance. Based on these observations, the NOEC for weight gain is 1836

mg a.i./kg feed.

LC50 > 5000 mg a.i./kg feed

NOEC (weight change) = 1836 mg a.i./kg feed

Conclusion: LC50 > 5000 mg a.i./kg feed (> 1118.95 mg a.i./kg body weight/day)

Chronic toxicity

Flag: Key study


Species: Bobwhite Quail (Colinus virginianus)

Type of exposure: 25 weeks, diet

Endpoint: NOEC

Value: 1000 mg a.i./kg feed (90.2 mg a.i./kg bw).

Reference: Kasugamycin technical: reproductive toxicity test with Northern Bobwhite (Colinus virginianus)

following OPPTS 850.2300 and OECD 206. JM Stafford. Study number 13917.4101. 18/03/2009

Klimisch score: 1

Amendments/deviations: no deviations that have a negative impact on the results or interpretation of the

study

GLP: yes

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September 2013

Test guidelines: OECD 206 (1984); OPPTS 850.2300 (draft, 1996)

No/sex/group: 18

Dose levels: 0, 250, 500 and 1000 mg/kg feed (nominal, mean measured are within ± 20% of nominal so the

nominal were used). The corresponding daily exposure by bird was 23.9; 45.0 and 90.2 mg a.i./kg bw.


Study summary: The objective of this study was to evaluate the effects of Kasugamycin technical via the diet

to Bobwhite Quail over a period of 25 weeks (10 weeks prior to photo-stimulation, approximately 5 weeks of

photo-stimulation and 10 weeks of egg collection). Adult parameters included health, body weight and feed

consumption. In addition, reproductive parameters (i.e. number of eggs laid, eggshell integrity and thickness,

fertility, viability, hatch rates, offspring survival and weight) were evaluated.

Average dietary concentration as measured in diet verification and homogeneity samples yielded mean

recoveries of 81.9% to 103% of nominal. Since measured concentrations were within ± 20% of nominal, per

guideline requirements, the results of this study are based on nominal concentrations.

The following acceptance criteria were met as required by the protocol: mortality in the control was less than

10% (8%), the average number of 14-day survivors per hen per set in the control was greater than 12 (44),

the average egg shell thickness for the control group was at least 0.19 mm (0.195 mm), the average

measured concentrations for the three treatment levels were greater than 80% of nominal concentrations

(81.9 to 103%).

No treatment related mortality occurred during the study and no overt signs of toxicity were detected during

behavioral observations or post-mortem examinations of the adults. No treatment related statistically

significant mean differences were detected among treatment groups in any of the adult data parameters

monitored during this test. One statistically significant difference was noted in egg production parameters.

The 500 mg a.i./kg feed group had a significantly lower ratio of 14-day old survivors of egg hatched

compared to control. The difference is primarily attributable to three different cages in that group, and is not

believed to be treatment related, as the effects were not observed at 1000 mg a.i/.kg feed, the NOEC was

1000 mg a.i/.kg feed (90.2 mg a.i./kg bw).

Conclusion: NOEC = 1000 mg a.i./kg feed (90.2 mg a.i./kg bw).

Flag: Key study


Species: Mallard duck (Anas platyrhynchos)

Type of exposure: 21 weeks, diet

Endpoint: NOEC

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September 2013

Value: 1000 mg a.i./kg feed (101.4 mg a.i./kg bw).

Reference: Kasugamycin technical: reproductive toxicity test with the Mallard (Anas platyrhynchos) following

OPPTS 850.2300 and OECD 206. JM Stafford. Study number 13917.4100. 18/03/2009

Klimisch score: 1

Amendments/deviations: no deviations that have a negative impact on the results or interpretation of the

study

GLP: yes

Test guidelines: OECD 206 (1984); OPPTS 850.2300 (draft, 1996)

No/sex/group: 16

Dose levels: 0, 250, 500 and 1000 mga.i./kg feed (nominal, mean measured are within ± 20% of nominal so

the nominal were used). The corresponding daily exposure by bird was 23.8; 48.5 and 101.4 mg a.i./kg bw.


Study summary: The objective of this study was to evaluate the effects of Kasugamycin technical via the diet

to Mallard duck over a period of 21 weeks (10 weeks prior to photo-stimulation, approximately 1 week of

photo-stimulation and 10 weeks of egg collection). Adult parameters included health, body weight and feed

consumption. In addition, reproductive parameters (i.e. number of eggs laid, eggshell integrity and thickness,

fertility, viability, hatch rates, offspring survival and weight) were evaluated.

Average dietary concentration as measured in diet verification and homogeneity samples yielded mean

recoveries of 81.3% to 108% of nominal. Since measured concentrations were within ± 20% of nominal, per

guideline requirements, the results of this study are based on nominal concentrations.

The following acceptance criteria were met as required by the protocol: mortality in the control was less than

10% (0%), the average number of 14-day survivors per hen per set in the control was greater than 12 (37),

the average egg shell thickness for the control group was at least 0.34 mm (0.343mm), the average

measured concentrations for the three treatment levels were greater than 80% of nominal concentrations

(81.3 to 108%).

No treatment related mortality occurred during the study and no overt signs of toxicity were detected during

behavioral observations or post-mortem examinations of the adults. No treatment related statistically

significant mean differences were detected among treatment groups in any of the adult data parameters

monitored during this test. One statistically significant difference was noted in egg production parameters.

The 250 mg a.i./kg feed group had a significantly higher number of cracked eggs compared to control. The

difference is primarily attributable to one cage in that group, and is not believed to be treatment related as

there were no significant effects at the 2 higher concentrations. Therefore, the NOEC was 1000 mg a.i/.kg

feed (101.4 mg a.i./kg bw).

Conclusion: NOEC = 1000 mg a.i./kg feed (101.4 mg a.i./kg bw).

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September 2013

General conclusion about ecotoxicity to birds

Kasugamycin has a low acute, short-term and chronic toxicity to birds. On this basis and given the data for

mammalian toxicity, no classification 9.3 is triggered for kasugamycin.

Ecotoxicity to bees

Laboratory tests (acute oral and dermal)

Flag: Key study


Species: Apis mellifera

Type of exposure: 48-hr exposure by contact and acute oral

Endpoint: LD50

Value: > 100 µg a.i./bee by oral and contact exposure

Reference: Kasugamycin technical: laboratory acute oral and contact toxicity test with the honeybee Apis

mellifera. KM. Nienstedt. Study number 1081.001.265. 30/08/2005

Klimisch score: 1


GLP: yes

Test guidelines: EPPO guideline 170 (1992); OECD 213 and 2014 (1998); Japan MAFF 12 Nohsan No 8147

(2000)

No/sex/group: 3 replicates of 10 bees per treatment

Dose levels: 100 µg a.i./bee by oral and contact exposure


Study summary: In the oral test, the food solution containing the test substance was completely consumed in

all cages. The mortality 48-hr after oral exposure was 6.7% in both the control and treatment groups. A total

of 3.3% of lethargic bees were observed on day 1 in the kasugamycin treatment group.

The mortality after 48-hr of contact exposure was 0% in all groups (control, solvent control and

kasugamycin). No lethargic bees were observed.

Therefore the LD50 for both oral and contact exposures is > 100 µg a.i./bee.

Conclusion: LD50 > 100 µg a.i./bee by oral and contact exposure

General conclusion about ecotoxicity to bees

No 9.4 classification is triggered for kasugamycin based on the acute oral and contact study on bees.

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September 2013

No other study has been provided in particular, no study on the effects on bee larvae is available.

Terrestrial invertebrate toxicity

Laboratory tests

Flag: Key study


Species: Pardosa spp

Type of exposure: direct application on quartz sand

Endpoint: LR50

Value: > 180 g a.i./ha

Reference: Kasugamycin technical: acute toxicity test with spiders, Pardosa spp. (Araneae: Lycosidae). KM.

Nienstedt. Study number 1081.001.272. 24/10/2005

Klimisch score: 1


GLP: yes

Test guidelines: Heimbach et al (2000): Guidelines to evaluate side-effects of plant protection products to

non-target arthropods. IOBC, BART and EPPO Joint Initiative.

No/sex/group: 15

Dose levels: 60 and 180 g a.i./ha


Study summary: Test organisms were exposed to direct application on quartz sand. Treatments were

sprayed in 400 L/ha. Mortality after 14 days, food consumption and behavior were recorded.

No mortality was observed in the control and at 60 g a.i./ha. At 180 g a.i./ha, mortality was 3.3% (not

statistically significant).

No adverse behavioral effects were observed on the spiders in the control and the kasugamycin treatment

groups.

The food consumption was reduced by 6.4 and 8.7% with respect to the control for 60 and 180 g a.i./ha

respectively. This reduction is statistically significant at the highest application rate.

Conclusion: LR50 > 180 g a.i./ha

Flag: Key study

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September 2013


Species: Typhlodromus pyri

Type of exposure: 7 days, dry residues on glass plates

Endpoint: LR50

Value: 102.0 g a.i./ha (95% confidence limits : 63.6-235.9)

Reference: Kasugamycin Technical – Dose response toxicity test with the predatory mite Typhlodromus pyri

Scheuten (Acari: Phytoseiidae). KM Nienstedt. Study number 1081.001.568 (29/08/2005)

Klimisch score: 1

Amendments/Deviations: no deviation that had a negative impact on the results or interpretation of the study

GLP: yes

Test guidelines: Blumel et al. 2000

No/group: 5 replicates of 20/ control, 4 replicates of 20/ test concentration

Dose levels: 13.5 – 27 – 54 – 108 – 216 g a.i./ha


Study summary: The objective of this study was to determine the potential effects of Kasugamycin technical

applied on glass plates on Typhlodromus pyri. The application rates were 13.5 – 27 – 54 – 108 – 216 g

a.i./ha, a toxic reference (dimethoate) was also included in the study.

On days 7, 9, 11 and 14, the numbers of dead, alive or missing mites were recorded as well as the sex of

adults, and numbers of eggs and larvae.

The LR50 obtained was 102.0 g a.i./ha (95% confidence limits : 63.6-235.9). A statistically significant

reduction of the number of eggs laid per female of 92.1, 97.3 and 90.2% was also observed at 13.5, 27 and

54 g a.i./ha, respectively.

The validity criteria of the guideline (mortality % in the control and reference, and reproduction rate of the

control) were met.

Conclusion: LR50 = 102.0 g a.i./ha. The reproduction of Typhlodromus pyri is impacted at sub-lethal

doses.


Test substance: kasugamycin technical

Species: Aphidius colemani

Type of exposure: Spray on filter paper and on leaves infested by aphids (hosts for the wasp)

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September 2013

Endpoint: LD50 at 48 h

Value: > 1000 ppm

Reference: Acute toxicity to Aphidius colemani of Kasugamycin. Y Sato. No study number available.

(15/11/2001)

Klimisch score: 3 (no guideline followed, not GLP, report of very poor quality)


GLP: no

Test guidelines: none

No/group: 3 replicates of 10

Dose levels: 100 and 1000 ppm

Analytical measurements: no

Study summary: The acute toxicity of Kasugamycin technical on the parasitic wasp Aphidius colemani was

studied by spraying filter paper and eggplant leaves infested by aphids with solution of 100 or 1000 ppm of

the test substance. The parasitic wasps were placed in Petri dishes containing the filter paper and one

treated leave. Mortality was assessed after 24 and 48 hours.

No mortalities were observed at 24 and 48 hours.

Conclusion: LD50> 1000 ppm



Species: Misumenops tricuspidatus (crab spider)

Type of exposure: direct spray on spiders

Endpoint: LD50 at 48 hours

Value: > 1000 ppm

Reference: Acute toxicity to Misumenops tricuspidatus of Kasugamycin. Y Sato. No study number available.

(15/11/2001)



GLP: no

Test guidelines: none followed

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September 2013

No/group: 20 replicates of 1 female / concentration



Study summary: The acute toxicity of Kasugamycin on the crab spider Misumenops tricuspidatus was

studied after direct spray with solution of 100 or 1000 ppm of the test substance. The spiders were placed

individually in a test tube containing treated rice seedling and 5 preys (leafhopper) and observed for 48

hours. Mortality was assessed after 24 and 48 hours.


Conclusion: LD50> 1000 ppm



Species: Chrysoperla carnea (lacewing) larvae

Type of exposure: direct spray on insects

Endpoint: LD50 at 48 hours

Value: > 1000 ppm

Reference: Acute toxicity to Chrysoperla carnea of Kasugamycin. Y Sato. No study number available.

(15/11/2001)



GLP: no

Test guidelines: none followed

No/group: 3 replicates of 10 insects



Study summary: The acute toxicity of Kasugamycin technical on the lacewing larvae Chrysoperla carnea was

studied after direct spray with solution of 100 or 1000 ppm of the test substance. The lacewings were placed

in a plastic box containing one treated eggplant leave infested by aphids and observed for 48 hours.

Mortality was assessed after 24 and 48 hours.


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September 2013

Conclusion: LD50 > 1000 ppm

General conclusion about ecotoxicity to non-target arthropods

Only 2 valid studies were provided on predatory mites and on Pardosa spiders. Pardosa spiders are known

to be less sensitive than the recommended indicator species (Typhlodromus pyri and Aphidius rhopalosiphi).

Therefore this species is not the most suitable for the risk assessment.

The predatory mites are more sensitive to kasugamycin with a LR50 of 102 g ai/ha which corresponds to the

application rate recommended in New Zealand.

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September 2013

Physico-chemical properties of the formulation

Table 15: Physical and chemical properties of Kasumin

Property Test method Klimisch Score

(1-4) Reference

Colour Dark Blue Green

Physical state Liquid

Odour None

Oxidizing properties None

pH

2.5 – 3.0

4.03 (1%

solution) 2

Application form


registration decision

PRD2012-30 Kasugamycin.

27/12/2013

Explosive properties

The product does

not contain

ingredients which

are potentially

explosive.

2


registration decision

PRD2012-30 Kasugamycin.

27/12/2013

Relative Density 1.01 – 1.03 2 Application form

Viscosity No information

Surface Tension 29 – 35 K dyn/cm 2 Application form

Water Solubility Yes 2 Note formulation is water-

based.

Solvent Solubility

(20°C) No information

Flammability No information

Auto flammability No information

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September 2013

Mammalian toxicology - Robust study summaries for the formulation

Acute toxicity [6.1]

Acute Oral Toxicity [6.1 (oral)]

Type of study: Acute oral toxicity in the rat

Flag: Key study

Test substance: Kasumin liquid (found on assay to contain 2.02% Kasugamycin

Endpoint: LD50 limit test in rats, signs of toxicity

Value: > 5000 mg/kg bw/day.

Reference: Cuthbert, J. A.; and Jackson, D.; 1992. Kasumin Liquid: Acute Oral Toxicity (Limit) Test in rats.

IRI Project No 553046 (Report No 9017). Inveresk Research International, Tranent, EH33 2NE, Scotland,

UK.

Klimisch Score: 1

Amendments/Deviations: The test substance was stored at ambient temperature for a week, but thereafter

refrigerated at 4°C as intended.

GLP: Yes

Test Guidelines: OECD No 401 (1981)

Species: Rat

Strain: Sprague-Dawley

No/Sex/Group: 5 (males and females)


Exposure Type: Oral gavage

Study summary: There were no deaths following a single orally administered dose of 5000 mg/kg bw to 5

male and 5 female rats. No clinical signs were noted and no abnormalities were seen at necropsy. Body

weight gains were acceptable.

Conclusion: The substance does not trigger classification for acute oral toxicity.

Acute Dermal Toxicity [6.1 (dermal)]

Type of study: Acute dermal toxicity in the rat

Flag: Key study

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September 2013

Test Substance: Kasumin liquid (found on assay to contain 2.02% Kasugamycin



Reference: Cuthbert, J. A.; and Jackson, D.; 1992. Kasumin Liquid: Acute Dermal Toxicity (Limit) Test in

rats. IRI Project No 553046 (Report No 9019). Inveresk Research International, Tranent, EH33 2NE,

Scotland, UK.

Klimisch Score: 1


refrigerated at 4°C as intended. One animal (No 20) was under dosed due to a calculation error so that this

animal was only dosed at circa 1740 mg/kg bw. [The study authors claimed that since none of the other

animals died, this does not invalidate the study.]

GLP: Yes


Species: Rats

Strain: Sprague-Dawley

No/Sex/Group: 5


Exposure Type: Dermal semi-occluded patch

Study Summary: There were no deaths. Clinical signs noted 4 hours (only) after dosing included red nasal

and ocular discharge and low body temperature in two males (No 11 and 15), while all females (No16 -20)

displayed red nasal discharge at 4 hours. [The degree of temperature depression was not stated

numerically.]

Body weight gains were acceptable for male, but were low for females giving a mean body weight gain over

the 14 days of 6 g (SD of ±4 g). [EPA staff note that in the acute oral study the female rats gained 43 ±8 g in

the 14 day period.]

The study author report the LD50 as > 2000 mg/kg bw

Conclusion: The substance triggers classification of 6.1E for acute dermal toxicity.

Comment: EPA staff note there were toxicological signs at the 2000 mg/kg bw level, but no deaths.

Classification as 6.1E for dermal toxicity is considered appropriate since the degree of reduced body weight

gain in female animals is considered toxicologically significant.

Acute Inhalation Toxicity [6.1 (inhalation)]

Type of study: Acute inhalation toxicity in the rat

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September 2013

Flag: Key study

Test Substance: Kasumin 2L (reported to contain 2.15% kasugamycin.

Endpoint: LC50, signs of toxicity

Value: > 4.89 mg/L

Reference: Shepherd, N; 2001. Kasumin 2L: Single Exposure (Nose only) Toxicity Study in the Rat. Report

No 1442/8-D6154. Covance Laboratories Ltd, Otley Road, Harrogate, North Yorkshire HG3 1PY, England.

Klimisch Score: 1

Amendments/Deviations: The study protocol for gravimetric assessment of the atmospheric concentration of

the substance was not used in preference to trapping the substance into a liquid medium. On a few

occasions the temperature and relative humidity were outside the protocol limits. [EPA staff note that in this

study the test substance Kasumin 2L was stored at room temperature in the dark. This supports the view

that the earlier oral and dermal studies would not have been impacted by the lack of refrigeration of the test

substance, as was claimed in the other reports.]

GLP: Yes

Test Guidelines: OECD No 403 (1981). US EPA Health Effects Test Guideline OPPTS 870.1300 Acute

inhalation toxicity (1998).

Species: Rat

Strain: Crl: WI (GIx/BRL/Han) BR

No/Sex/Group: 5 (males and females)

Dose Levels: 4.892 mg Kasumin 2L/L (105.1643 µg kasugamycin/L) [The sample collected at one time point

was discarded as an outlier due to a procedural analytical error.]

Exposure Type: 4 hour inhalation exposure via nose only. The mass median aerodynamic diameter of the

particles was 1.86 µm, with a geometric standard deviation of 2.13.

Study Summary: One animal (no7) was removed from the study on Day 14 due to its moribund condition.

Clinical signs were observed following exposure which were attributed to treatment included gasping,

labored breathing, noisy breathing, wheezing, stained fur, hunched [posture] and animals being thin. Several

of these signs persisted through until Day 10 of observation, while wheezing persisted until Day 15. The

condition of female animals 7 and 9 deteriorated towards the end of the observation period (EPA staff, ie the

end of the study) and animal 7 was removed (ie euthanased) for humane reasons. Minor clinical staining of

the eyes on Day 1 was attributed to restraint not the exposure to the test article.

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September 2013

All exposed animals lost weight on Day 2-3, but by Day 5 the losses stopped and all but one animal gained

weight. The body weight gains of these animals continued to increase for the remainder of the study but

were reduced in comparison with that expected for animals of this strain and age.

There was not effect on lung weight in animals at termination but animal 7 had increased lung weight.

The only finding at necropsy attributed to treatment was the gaseous distension and dark lung in animal no 7

(attributed to agonal changes, in this moribund animal).

The study authors proposed that as there was one death at 4.892 mg/L the LC50 can be predicted to be

above 5 mg/L.

Conclusion: The substance triggers classification as 6.1E for acute inhalation.

Comments: In view of the signs of toxicity including one death that appears to be test substance-related

classification as 6.1E (inhalation) is appropriate.

Skin Irritation [6.3/8.2]

Type of study: Skin irritation/corrosion in the rabbit

Flag: Key study



Value: 0.0 for both erythema and oedema

Reference: Cuthbert, J. A. et al.; 1992. Kasumin Liquid: Primary Skin Irritation Index Test in rabbits. IRI

Project No 553046 (Report No 9020). Inveresk Research International, Tranent, EH33 2NE, Scotland, UK.

Klimisch Score: 1



GLP: Yes

Test Guidelines: OECD No 404 (1981). US EPA guideline, Subdivision F, 81-5 (1982)

Species: Rabbit


No/Sex/Group: 6 (males)

Dose: 0.5ml under gauze

Study Summary: There were no signs of erythema or oedema throughout the study.

Conclusion: The substance does not trigger classification as a skin irritant or corrosive.

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September 2013

Eye Irritation [6.4/8.3]

Type of study: Eye irritation/corrosion

Flag: Key study


Endpoint: Mean Draize Score (24, 48 and 72 hours)

Value: Non-rinsed group: conjunctival redness = 4/18 = 0.22; conjunctival chemosis = 0.0 Cornea and iris

scores were 0.0.

Rinsed group: scores for all endpoints were 0.0.

Reference: Cuthbert, J. A. et al.; 1992. Kasumin Liquid: Primary Eye Irritation Test in rabbits. IRI Project No

553046 (Report No 9021). Inveresk Research International, Tranent, EH33 2NE, Scotland, UK.

Klimisch Score: 1

Amendments/Deviations: The test substance was stored at ambient temperature for a week but thereafter


GLP: Yes

Test Guidelines: OECD No 405 (1987). US EPA Guidelines Subdivision F 8104 (1982)

Species: Rabbits


No/ Sex/Group: 9 (6 without rinsing, and 3 with rinsing).

Dose: 0.1 ml

Study Summary: All treated rabbits had moderate conjunctival responses at 1 hour, with slight redness

persisting in one rabbit in the non-rinsed group till 48 hours. One rabbit in the non-rinsed group showed slight

iritis 1 hour post-instillation. In the rinsed group, slight discharge persisted in one animal until 48 hours, and

two rabbits showed signs of light iritis at 1 hour. Full recovery occurred by 72 hours.

Conclusion: The substance does not trigger eye irritation/corrosion.

Contact Sensitization [6.5]

Type of Study: Contact sensitisation in the guinea pig

Flag: Key study


Endpoint: Sensitisation response

Value: Negative

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September 2013

Reference: Cuthbert, J. A.; and Jackson, D.; 1992. Kasumin Liquid: Buehler Sensitisation Test in Guinea

Pigs. IRI Project No 553046 (Report No 9022). Inveresk Research International, Tranent, EH33 2NE,

Scotland, UK.

Klimisch Score: 1

Amendments/Deviations: The test substance was stored at ambient temperature for a week but thereafter


GLP: Yes


Species: Guinea pigs

Strain: Dunkin-Hartley albino

No/Sex/Group: 20 in the test and control groups (all females)

Type of Exposure: Semi occluded patch test.

Study Summary: The dose range finding study used 100, 50, 25 and 10% of the substance in distilled water.

As there was no reaction at all concentrations, 100% was used in the subsequent test system for both

induction and challenge phases.

There was no reaction during the induction or challenge phases using 100% of the test substance. Data

were presented validating the study in a positive control (2,4 dinitrochlorobenezene).

Conclusion: The substance does not trigger classification as a contact sensitiser.

General conclusion about acute toxicity

The substance does not trigger classification for acute oral toxicity, skin or eye irritancy or contact

sensitisation, but triggers classification of 6.1E for both dermal and inhalation routes.

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September 2013

Ecotoxicity - Robust study summaries for the formulation

Aquatic toxicity

Fish acute toxicity (Freshwater species)

Flag: key study

Test substance: Kasumin 2L

Species: Common carp (Cyprinus carpio)

Type of exposure: 96 hr static. (control medium was renewed at 48Hr)

Endpoint: LC50

Value: 120 mg formulation/L

Reference: A 96-hr acute toxicity study of Kasumin 2% liquid with Common Carp. M. Anai. Study number

94341. 22/06/2007

Klimisch score: 2 no analytic measurement of the concentration but the validity criteria of the guideline were

met.


GLP: yes

Test guidelines: OECD 203 (1992). Japan MAFF subdivision 2-7-1-1 (2005)

No/group: 10

Dose levels: 77.0; 100; 130; 169; 220 mg formulation /L


Study summary: Groups of 10 common carps were exposed to Kasumin 2L for 96-hr in static conditions at

concentrations ranging from 77.0 to 220 mg/L. No analytic determinations of the concentrations were

performed. 100% of mortality was observed at 169 mg/L and above while 100 mg/L did not produce any

mortality. Abnormal findings in the treated groups were: loss of equilibrium, lethargy, reduced activity, and

reduced respiration.

Neither mortality nor abnormal findings were observed in the control group.

Conclusion: LC50 = 120 mg formulation/L corresponding to 2.4 mg ai/L. There is an obvious effect of

the formulation since the LC50 of the active ingredient are all above 110 mg ai/L.

Invertebrate acute toxicity (Freshwater species)

Flag: key study


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September 2013


Type of exposure: 48-hr static

Endpoint: EC50

Value: 152 mg formulation/L

Reference: A 48-hr acute immobilization study of Kasumin 2% liquid with Daphnia magna. T. Matsuura.

Study number 94340. 20/06/2007

Klimisch score: 2 no analytic measurement of the concentration but the validity criteria of the guideline were

met.


GLP: yes

Test guidelines: OECD 202 (2004), Japan MAFF subdivision 2-7-2-1 (2005)

No/sex/group: 20

Dose levels: 87.5; 114; 148; 192; 250 mg formulation/L


Study summary: Groups of 20 Daphnia magna were exposed to Kasumin 2L for 48-hr in static conditions at


performed. 100% of immobilization was observed at 250 mg/L while 114 mg/L did not produce any

immobilization.

Conclusion: EC50 = 152 mg formulation/L corresponding to 3.04 mg ai/L. There is an obvious effect of

the formulation since the EC50 of the active ingredient is above 90 mg ai/L.

Algae acute toxicity (Freshwater species)

Flag: key study


Species: Pseudokirchneriella subcapitata

Type of exposure: 72 hr static

Endpoint: ErC50 and EbC50

Value: ErC50 = 458 mg formulation/L (24-72hr) and EbC50 = 132 mg formulation/L (0-72hr)

Reference: Algae growth inhibition study of Kasumin 2% liquid with Pseudokirchneriella subcapitata. M. Seki.

Study number 94339 19/06/2007

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September 2013

Klimisch score: 2 no analytic measurement of the concentration. The factor between concentrations is 3.5

while the guideline requires 1.3 to 2.2. At the date of the study, an updated OECD guideline was available

but has not been followed.


GLP: yes

Test guidelines: Japan MAFF subdivision 2-7-7 (2005), OECD 201 (1984)

Concentration levels: 6.66; 23.3; 81.6; 286; 1000 mg formulation/L


Study summary: Pseudokirchneriella subcapitata was exposed to Kasumin 2L for 72-hr in static conditions at


performed. The following results were obtained: ErC50 =458 mg/L (24-72hr) and EbC50 = 132 mg/L (0-72hr)

Conclusion: ErC50 = 458 mg formulation /L (24-72hr) and EbC50 = 132 mg formulation /L (0-72hr)

corresponding to 9.16 and 2.64 mg ai/L, respectively. There is an effect of the formulation since the

EC50 of the active ingredient is 14 mg ai/L.

General conclusion about aquatic ecotoxicity

No 9.1 classification is triggered for Kasumin 2L regarding the effects on aquatic organisms.

Soil toxicity

Non-target plants acute toxicity (vegetative vigour)

Flag: Key study


Species: Allium cepa (Onion), Lolium perenne (Ryegrass), Triticum aestivum (Wheat), Zea mays (Corn),

Beta vulgaris (Sugarbeet), Linum usitatissiumum (Flax), Glycine max (Soybean), Lactuca sativa (Lettuce),

Lycopersicon esculentum (Tomato), Raphanus sativus (Radish)

Type of exposure: foliar application

Endpoint: ER25

Value: ER25 > 4675 g/ha of formulated product (95.18 g a.i./ha)

Reference: Kasumin 2L: A toxicity test to determine the effects of the test substance on vegetative vigour of

10 species of plants. Report number 443-117. JR Porch et al. 17/12/2009

Klimisch score: 1

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September 2013


GLP: yes

Test guidelines: US EPA Series 850 Ecological effects test guidelines OPPTS number 850.4150

No/group: 6 replicates of 5 individual plants

Dose levels: 64 fluid ounces Kasumin 2L per acre corresponding to approximately 4675 g/ha of formulated

product (95.18 g a.i./ha)

Analytical measurements: the concentration in the spray mixture was checked by HPLC before spray

Study summary: For each plant species tested, young plants were exposed to a single concentration of

Kasumin 2L equivalent to the maximum single labeled application rate by foliar spray. The study duration

was 21 days.

There were no treatment-related effects on the height, dry weight or survival of any of the ten plant species

tested. Treatment group mean reductions of 25% or greater relative to the negative control means were not

observed for any endpoints in any of the plant species (maximum effect was 9% for dry weight of tomato). In

addition, treatment group means were not determined to be significantly different from any of the respective

negative control means, with two exceptions. The Day 7 treatment group height means of Lactuca sativa,

and Raphanus sativus were determined to be significantly different from respective negative control means;

however, the difference in height between the two means was slight (< 2 cm). Day 14 and Day 21 treatment

group height means of L. sativa and R. sativus were determined not to be significantly different from their

respective negative control means. Signs of toxicity, such as necrosis, chlorosis, and leaf curl were observed

on some plants. Signs were noted with greater frequency in the treatment group for a few species.

Conclusion: Application of the Kasumin 2L at the maximum label rate of 4675 g/ha (95.18 g a.i./ha)

resulted in no significant effect (> 25%). ER25 > 4675 g/ha of formulated product (95.18 g a.i./ha)

Non-target plants acute toxicity (seedling emergence)

Flag: Key study


Species: Allium cepa (Onion), Lolium perenne (Ryegrass), Triticum aestivum (Wheat), Zea mays (Corn),

Beta vulgaris (Sugarbeet), Linum usitatissiumum (Flax), Glycine max (Soybean), Lactuca sativa (Lettuce),

Lycopersicon esculentum (Tomato), Raphanus sativus (Radish)

Type of exposure: spray application on soil

Endpoint: ER25

Value: ER25 > 4675 g/ha of formulated product (95.18 g a.i./ha)

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September 2013

Reference: KASUMIN 2L: A toxicity test to determine the effects of the test substance on seedling

emergence of ten species of plants. John R. Porch, et al. Study No.443-116. 07/10/2009

Klimisch score: 1


GLP: yes

Test guidelines: OPPTS Number 850.4100 (1996)

No/group: 4 replicates of 10 plants per group

Dose levels: 95.18 g a.i./ha (4675 g formulation/ha)

Analytical measurements: HPLC/UV used to measure the concentration in the spray mixture

Study summary: The study was conducted to determine the effects of the test substance on the seedling

emergence, survival, growth and condition of ten species of plants. Application of Kasumin 2L at the

maximum label rate of 95.18 g a.i./ha resulted in no adverse effects on emergence, survival, or condition of

the ten terrestrial plant species tested. In addition, effects of the application of 25% or greater were not

observed on the height and dry weight of Z. mays, B. vulgaris, G. max, L. esculentum, and R. sativus.

Growth results for A. cepa (26% reduction of dry weight), L. perenne (21% reduction of dry weight), T.

aestivum (37% reduction of dry weight), L. sativa (189% increase of dry weight), and L. usitatissimum (82%

increase of dry weight) appear to reflect a correlation with the timing of emergence rather than a treatment

related effect.

Conclusion: Application of the Kasumin 2L at the rate of 4675 g formulation/ha (95.18 g a.i./ha)

resulted in no significant effect (> 25%). ER25 > 4675 g/ha of formulated product (6.23 mg

formulation/kg dry weight soil).

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September 2013

Appendix B: Risk assessment

1. To facilitate the assessment of risks, the applicant and the staff identified the most common potential

sources of risk to the environment and to human health and safety through release, spillage or exposure

throughout the lifecycle of the substance, assuming no controls or regulations are complied with. These

are tabulated in Table 1.

2. Staff carried out quantitative and qualitative assessments to assess risks to human health and the

environment from the use of Kasumin. The results of this are shown in Section 6 of this report.

3. The process by which the risk assessment of substances is undertaken is specified in the

Methodology12

. Guidance on risk assessment is provided on the EPA website13

Table 1: Potential sources of risks associated with hazardous substances

Lifecycle Activity Associated Source of Risk

Manufacture / Import

An incident during the manufacture or importation of the substance

resulting in spillage and subsequent exposure of people or the

environment to the substance.

Packing An incident during the packing of the substance resulting in spillage and

subsequent exposure of people or the environment to the substance.

Transport or storage

An incident during the transport or storage of the substance resulting in

spillage and subsequent exposure of people or the environment to the

substance.

Use

Application of the substance resulting in exposure of users or bystanders

or the environment; or an incident during use resulting in spillage and

subsequent exposure of users or the environment to the substance.

Disposal Disposal of the substance or packaging resulting in exposure of people or

the environment to the substance.

12 http://www.epa.govt.nz/publications/methodology.pdf 13 http://www.epa.govt.nz/Publications/ER-TG-05-02-03-09-(Decision-Making).pdf

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September 2013

Human health risk assessment

Quantitative worker (operator) risk assessment

Critical endpoint definition

Deriving an AOEL 14

Key

systemic

effect

NOAEL

(LOAEL)

mg/kg

bw/day

Uncertainty

factors

Absorption

factor15

AOEL

mg/kg

bw/day

Justification

13 week rat:

kidney

17.73 mg/kg

bw/day in

males

100 0.05 0.009 Note that in the rat less than 5%

of the dose is absorbed.

13 week

dog:

mouth/tongu

e changes.

10.59 mg/kg

bw/day in

males

- - -

These data are not confirmed by

1 year dog study so should not be

used.

13 week

mouse:

mortality,

changes in

kidney, anus

and testes

135.4 mg/kg

bw/day in

males

100 0.05 0.068

Gives a higher value than that

from the rat study: the rat study

should be used to derive the

AOEL.

14

The toxicological endpoint used for assessment of occupational (worker) and re-entry worker risks is the AOEL (Acceptable Operator Exposure Level). The AOEL is the maximum amount of active substance to which the operator/re-entry worker may be exposed with a low probability of adverse health effects amongst the healthy worker sub-population, allowing for some margin of safety. AOELs describe the internal (absorbed) dose available for systemic distribution from any route of absorption and are expressed as internal (systemic) levels (mg/kg bw/day). They are derived by dividing the most appropriate NOAEL from relevant studies by one or more uncertainty (safety) factors selected on the basis of the extent and quality of the available data, the species for which data are available and the nature of the effects observed. An absorption factor may be applied to take into account the absorbed dose in the study where this is known (this is a percentage expressed as a factor);

15

100

n% Absorptio

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September 2013

Other inputs for human worker (operator) and re-entry exposure modelling16

Table 2: Derivation of dermal absorption value in humans

Active

Physical

form

Concentration

of each active

(g/L or g/kg)

Log

Kow

Maximum

application

rate (for each

active, for each

method of

application)

g a.i./ha

Dermal absorption

(%)

AOEL

mg/kg

bw/day Concentr

ate Spray

Kasugamycin Liquid 20 g/L No data 100g/ha 50% (No

data)

50% (No

data) 0.009

Comments on inputs for human worker (operator) exposure modelling input parameters:

No data presented on dermal absorption. The proportion absorbed following oral administration in the rat

toxicokinetics study is extremely low (< 5%), resulting in a low AOEL. EPA staff consider it may be

reasonable to assume dermal absorption will be low given the low oral absorption and taking into account

the large molecular weight of the substance. However, with no data a default value is used as the

assumption. EPA staff note that the Canadian PMRA assumed 100% dermal absorption in the absence of

dermal absorption data.

Output of human worker (operator) mixing, loading and application exposure modelling

Exposure Scenario Estimated operator exposure

(mg/kg bw/day) Risk Quotient

Boom

No PPE17 during mixing, loading and application 0.0635 7.05

Gloves only during mixing and loading 0.0295 3.28

Gloves only during application 0.0581 6.46

Full PPE during mixing, loading and application

(excluding respirator)

0.0016 0.18


(including respirator)

0.0016 0.18

Airblast

16

The Staff has undertaken an assessment of risks to operator health using the United Kingdom Pesticide Safety Directorate’s interpretation of the German BBA Model to estimate operator exposure. This model estimates the exposure of workers to a pesticide during mixing, loading and during spray application, in mg/kg person/day (http://www.pesticides.gov.uk/index.htm). The derived values consider both dermal and inhalation exposure routes. The Staff typically uses the geometric mean model. The BBA model provides for a range of different spray applications (tractor-mounted/trailed sprayers and hand-held sprayers) and formulation types (liquid, wettable powder and wettable granule). Additionally, the BBA model also allows flexibility to vary protective clothing (hands, head and body). 17

Full” PPE includes: gloves, hood/visor, coveralls, and heavy boots during application. The model only provides for use of gloves at mixing loading.

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September 2013

No PPE during mixing, loading and application 0.0796 8.85





0.0035 0.39



0.0032 0.36

Backpack - High Level Target

No PPE during mixing, loading and application 0.1758 19.53





0.0031 0.34



0.002580 0.29

Outcomes of the worker (operator) exposure assessment:

Risks are controllable by the use of PPE

Full PPE is recommended (gloves, hood/visor, coveralls, and heavy boots with or without a respirator)

during mixing, loading, and application in order to reduce the risks to an acceptable level

Outcomes of the re-entry exposure assessment:

Table 3: Re-entry exposure modeling18

Active

Internal (absorbed) dose available

for systemic distribution

(mg/kg bw/8 hours)

AOEL

(mg/kg

bw/day)

Risk Quotient

Kasugamycin

Boom

Airblast

Aerial

0.135

0.135

0.253

0.009

15.00

15.00

15.00

18

The Staff assessed the re-entry worker exposures using the generic exposure model for “Maintenance and harvesting activities: Dermal exposure” provided by the UK Health & Safety Executive chemical Regulation Directorate, on the following web site: http://www.pesticides.gov.uk/applicant_guide.asp?id=1246&link=%2Fuploadedfiles%2FWeb%5FAssets%2FPSD%2FRe%2Dentry%2520worker%2520guidance%5Ffinal%2520version%2Epdf.

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September 2013

A §77A re-entry control is recommended to reduce the level of re-entry risk.

Use of PPE for early crop entry is recommended as necessary.

The modelling suggests that a re-entry interval of 40 days is required if no PPE is worn or 30 days when

gloves are worn, but EPA staff consider this is a conservative estimate particularly due to use of the default

50% dermal absorption figure in the absence of specific data. Staff therefore recommend that a re-entry

interval is set for as long a duration (up to 30 days with gloves or 40 days with no PPE) as is reasonably

practicable.

Quantitative bystander risk assessment

Critical endpoints definition

The AOEL as derived above is used for the bystander risk assessment.

Output of human bystander mixing, loading and application exposure modelling19

Exposure Scenario

Estimated exposure of 15 kg

toddler exposed through

contact to surfaces 8 m from an

application area

Risk Quotient

Boom

High boom, fine droplets 3.47 0.3857

High boom, coarse droplets 0.55 0.0612

Low boom, fine droplets 1.17 0.1302

Low boom, coarse droplets 0.28 0.0310

Airblast

Airblast sparse orchard 10.03 1.1145

Airblast dense orchard 3.35 0.3721

Airblast vineyard 0.46 0.0516

Aerial – agriculture

Swath width 20 m, Med-coarse droplet size 4.79 0.5327

Swath width 20 m, coarse- v. coarse droplets 3.54 0.3937

19

Exposure is estimated using the equations from the UK Heath & Safety Chemical Regulation Directorate which account for dermal exposure, hand-to-mouth exposure and object-to-mouth exposure (http://www.pesticides.gov.uk/uploadedfiles/Web_Assets/PSD/Bystander%20exposure%20guidance_final%20version.pdf Accessed 27/01/2010a). In addition, incidental ingestion of soil is taken into account using a modified exposure equation from the United States Environmental Protection Agency (USEPA, 2007, Standard Operating Procedures (SOPs) for Residential Exposure Assessments, Contract No. 68-W6-0030, Work Assignment No. 3385.102). Spray drift is estimated using models specific to the type of application equipment. For pesticides applied by ground boom or air blast sprayer, the AgDrift model is used. Spray drift deposition from aerial application is estimated using the AGDISP model along with appropriate New Zealand input parameters

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September 2013

Swath width 20 m, extremely coarse droplets 2.39 0.2659

Swath width 24 m, v. fine-fine droplets 14.27 1.5861

Swath width 24 m, fine-med. droplets 7.09 0.7876

Swath width 24 m, med.-coarse droplets 4.75 0.5279

Outcomes of the bystander exposure assessment

The risks to bystanders are acceptable, except for airblast in a sparse orchard and aerial use in

agriculture with very fine/fine droplet size for a bystander exposure risk is estimated to apply.

While the RQ value for bystanders from airblast application in a sparse orchard is slightly above the level

of concern, staff note that this is likely to be an over estimate due to the use of conservative default

values. It is therefore considered that this use scenario is unlikely to present a significant risk of adverse

effects.

It is recommended that a control relating to droplet size for aerial application be included to address this

potential bystander risk.

Summary and conclusions of the human health risk assessment

The risks to operators are acceptable provided workers (operators) use full PPE with or without a respirator

during mixing, loading and application.

The risks to re-entry workers are identified as high. EPA staff consider that the estimates are likely to

substantially over estimate re-entry risks, primarily due to the use of the default 50% dermal absorption value

and a lack of information on dislodgeable foliar residues and foliar half-life. However, in the absence of

specific data it is recommended that a re-entry interval is established for as long a duration (up to 30 days

with gloves or 40 days with no PPE) as is reasonably practicable..

The risks to bystanders are acceptable provided, in the case of aerial application, fine or very fine spray

systems are not used, so a recommendation is made for a control on the spray size.

While the RQ value for bystanders from airblast application in a sparse orchard is slightly above the level of

concern, this is likely to be an over estimate and it is therefore considered that this use scenario is unlikely to

present a significant risk of adverse effects.

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September 2013

Environmental risk assessment

8.3. Environmental fate and ecotoxicity list of endpoints used for risk

assessment20 purpose and classification

8.3.1. Active ingredient

Table 4: Persistency in aquatic and terrestrial environment (values in bold are used for the risk assessment)

Test type Test method Test results - Days

Klimisch

score

(1-4)

Reference

Ready biodegradation No data

Aerobic water/sediment (DT50) OECD 308 18.2 and 28.6 1 Study 1819W

(2009)

Aqueous photolysis half-life

(DT50)

Japan MAFF

and US EPA 8.2 (natural water) 1

Study 1442/22

(2003)

Hydrolysis half-life (DT50) at

25oC

EC C7

> 1 year at pH 4 and 5.

DT50 = 72.4 days at pH 7

and 10.8 days at pH 9

1 Study 1442/21

(2003)

Adsorption/desorption (Koc) a OECD 106 134 to 324. 1

Study 1820W

(2009)

Aerobic half-life in soil (DT50) -

laboratory US EPA 40.8 2

Study 6434-106

(1998)

Conclusion on persistency: Kasugamycin is not rapidly degradable in water (DT50 > 16 days) and

persistent in soil (DT50 > 30 days).

a All values except in sand.

20 Depending on the risk assessment the endpoint values used can differ from those listed in the presented tables.

Further explanation is presented in the appropriate paragraph under the environmental risk assessment section.

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September 2013

Table 5: Bioaccumulation

Table 6: Aquatic toxicity (only the key studies are mentioned below)

Test species Test

method

Test type

and

duration

Test

results –

mg a.i./L

Klimisch

score

(1-4)

Reference

Fish

(Rainbow trout) US EPA

96 hr,

static LC50 > 120 1 Study 13917.6114 (2009)

Invertebrates

(Daphnia magna) OECD 202

48 hr,

static

EC50 >

90.7 1 Study 1442/16 (2002)

Algae

(Anabaenaflos-aquae ) OECD 201 96hr static

ErC50 =

1.3 1 Study 13917.6111 (2009)

Aquatic plants (Lemna

gibba) OECD 221

7 days,

semi-static EC50 > 110 1 Study 13917.6107 (2009)

Conclusion on aquatic classification: 9.1 B Toxic for the aquatic organisms

Test type Test

method

Test

results

Klimisch

score

(1-4)

Reference

Partition coefficient

octanol/water Unknown < - 1.96 2

Health Canada - Proposed registration

decision PRD2012-30 (27/11/2012)

Fish

bioconcentration

test

No data

Conclusion on bioaccumulation: Not potentially bioaccumulative

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September 2013

Table 7: Soil toxicity

Test species Test method

Test type

and

duration

Test results –

mg a.i./kg soil

Klimisch

score

(1-4)

Reference

Earthworm OECD 207 14 days,

static LC50 > 1000 1

Study 1081.001.630

(2005)

N transformation in

soil

Norm of the

Netherlands

8 weeks,

static

Significant effects

at 5 kg ai/ha 2 Study 231795 (1998)

Conclusion on soil classification: Not triggered

Table 8: Terrestrial vertebrate toxicity

Test species Test

method

Test type

and

duration

Test

results

Klimisch

score

(1-4)

Reference

Mallard and

Bobwhite quail US EPA acute

LD50 >

2000 mg

a.i./kg/d

1 Study 13862.4102 (2006) and

13862.4103 (2006)

Mallard and

Bobwhite quail OECD 205

Dietary, 5

days

LC50 >

5000 ppm 1

Study 13862.4101 (2006) and

13862.4100 (2006)

Conclusion on terrestrial vertebrate classification: Not triggered (for mammalian results, see the

toxicology part)

Table 9: Terrestrial invertebrate toxicity

Test species Test

method

Test type

and

duration

Test

results -

µg a.i/bee

Klimisch

Score

(1-4)

Reference

Apis melifera OECD 213

and 214

Acute, 48

hr, contact

and oral

LD50 > 100 1 Study 1081.001.265 (2005)

Conclusion on terrestrial invertebrate classification: Not triggered

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September 2013

8.3.2. Formulation

Table 10: Aquatic toxicity

Test species Test

method

Test type

and

duration

Test results

Klimisch

score

(1-4)

Reference

Fish

(Common carp) OECD 203 96 hr static

LC50 = 120 mg

formulation/ L

(2.4 mg ai/L)

2 Study 94341 (2007)

Invertebrates

(Daphnia magna) OECD 202

48 hr,

static

EC50 = 152 mg

formulation/ L

(3.04 mg ai/L)

2 Study 94340 (2007)

Algae

(Pseudokirchneriella

subcapitata)

OECD 201 72 hr static

EbC50 = 132*

mg formulation/

L (2.64 mg ai/L)

2 Study 94339 (2007)

Conclusion on aquatic classification: Not triggered

* EbC50 was chosen instead of ErC50 because the validity criteria regarding the growth of the algae were not checked in

this study and there was no analytical measurement so as a precautionary approach the lowest value was used.

8.4. Risk assessment methodology

Methods used to assess environmental exposure and risks differ between environmental compartments

(Table ).

Table 11: Reference documents for environmental exposure and risk assessments

Environmental exposure Risk assessment

Aquatic organisms

(GEN)eric (E)stimated

(E)nvironmental (C)oncentration

Model Version 2.0 – 01 August

2002

AgDrift and EPA Software21

Overview of the Ecological Risk Assessment

Process in the Office of Pesticide Programs, U.S.

Environmental Protection Agency. Endangered

and threatened Species Effects Determinations –

23 January 2004

Sediment Guidance on information Guidance on information requirements and

21

The Staff used two different models for assessing the EEC and associated risks:

Generic Estimated Environmental Concentration Model v2 (GENEEC2) surface water exposure model (USEPA,

2001) estimates the concentration of substance in surface water which may arise as a result of surface runoff and

spraydrift.

To examine how buffer zones would reduce the active ingredient concentrations in receiving waters, the Staff used

the AgDRIFT®

model (developed under a cooperative Research and Development Agreement, CRADA, between the

EPA, USDA, US Forest Service, and SDTF). AgDRIFT®

incorporates a proposed overall method for evaluating off-

site deposition of aerial, orchard or ground applied pesticides, and acts as a tool for evaluating the potential of buffer

zones to protect sensitive aquatic and terrestrial habitats from undesired exposures. Calculations are made assuming

the receiving water is a 30 cm deep pond. The model is used to estimate the buffer zone that would reduce exposure

through spray drift to such a concentration that an acute risk quotient of 0.1 cannot be calculated. It is noted that

unlike GENEEC2, AgDRIFT®

model only considers transport by spray drift, input through runoff, volatilisation, etc will

pose additional risks.

129


September 2013

organisms requirements and

chemical safety assessment,

Chapter R.16: Environmental

Exposure Estimation, Version: 2 -

May 2010

chemical safety assessment, Chapter R.10:

Characterisation of dose [concentration]-response

for environment – May 2008

Soil organisms,

invertebrates

(macro-

invertebrates)

Soil persistence models and EU

registration. The final report of the

work of the Soil Modelling Work

group of FOCUS (FOrum for the

Co-ordination of pesticide fate

models and their USe) – 29

February 1997

SANCO/10329/2002 rev 2 final. Guidance

Document on terrestrial ecotoxicology under

Council Directive 91/414/EEC- 17 October 2002

Terrestrial

organisms,

invertebrates (non-

target arthropods)

Guidance document on regulatory testing and risk assessment procedures for plant

protection products with non-target arthropods. From ESCORT 2 Workshop – 21/23

March 2000

Terrestrial

vertebrates (birds)

Guidance of EFSA. Risk assessment to birds and mammals – 17 December 2009.

EFSA calculator tool - 200922

SANCO/4145/2000 final. Guidance Document on risk assessment for birds and mammals

under Council Directive 91/414/EEC- 25 September 2002

Secondary

poisoning and

biomagnification

Technical Guidance Document on

risk assessment in support of

Commission Directive 93/67/EEC

on Risk Assessment for new notified

substances, Commission

Regulation (EC) No 1488/94 on

Risk Assessment for existing

substances, Directive 98/8/EC of

the European Parliament and of the

Council concerning the placing of

biocidal products on the market –

Part II - 2003

Guidance of EFSA. Risk assessment to birds and

mammals – 17 December 2009

EFSA calculator tool - 2009

SANCO/4145/2000 final. Guidance Document on

risk assessment for birds and mammals under

Council Directive 91/414/EEC- 25 September

2002

8.5. Aquatic risk assessment

For Class 9 substances, irrespective of the intrinsic hazard classification, the ecological risk can be assessed

for a substance by calculating a Risk Quotient (RQ) based on an estimated exposure concentration. Such

calculations incorporate toxicity values, exposure scenarios (including spray drift, leaching and run-off,

application rates and frequencies), and the half-lives of the component(s) in water. For the aquatic

environment, the calculations provide an Estimated Environmental Concentration (EEC) which, when divided

by the L(E)C50 or a NOEC, gives a RQ acute or chronic.

( )

22

www.efsa.europa.eu/en/efsajournal/pub/1438.htm

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September 2013

If the RQ exceeds a predefined level of concern, this suggests that it may be appropriate to refine the

assessment or apply the approved handler control and/or other controls to ensure that appropriate matters

are taken into account to minimize off-site movement of the substance. Conversely, if a worst-case scenario

is used, and the level of concern is not exceeded, then in terms of the environment, there is a presumption of

low risk which is able to be adequately managed by such things as label statements (warnings, disposal).

The approved handler control can then be removed on a selective basis.

Levels Of Concern (LOC) developed by the USEPA (Urban and Cook, 1986) and adopted by EPA determine

whether a substance poses an environmental risk (Table ).

Table 12: Levels of concern as adopted by EPA New Zealand

Endpoint LOC Presumption

Aquatic (fish, invertebrates)

Acute RQ ≥ 0.5 High acute risk

Acute RQ 0.1 - 0.5 Risk can be mitigated through restricted use

Acute RQ < 0.1 Low risk

Chronic RQ ≥ 1 High chronic risk

Plants (aquatic and terrestrial)

Acute RQ ≥ 1 High acute risk

When available, data from the formulation will be used in the risk assessment.

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September 2013

8.5.1. GENEEC2 modelling

Calculation of expected environmental concentrations

The parameters used in GENEEC2 modelling are listed in Table .Error! Reference source not found.

Table 13: Input parameters for GENEEC2 analysis

Kasugamycin Ground based application on

kiwifruit

Application rate (kg/ha) 0.1

Application frequency 4

Application interval (days) 10

Koc 134

Aerobic soil DT50 (days)

40.8. As there is only one value,

the input parameter is 3 x 40.8

= 122.4*

Pesticide wetted in? no

Methods of application Airblast spray

‘No spray’ zone nil

Water solubility (ppm) 228000

Hydrolysis (DT50 in days) 72.4

Aerobic aquatic DT50 (days) 28.6

Aqueous photolysis DT50

(days) 8.2

*According to the GENEEC2 User’s manual recommendations (August 2001)

Output from the GENEEC2 model

The peak maximum Estimated Environmental Concentrations (EEC) for Kasugamycin when used in

kiwifruit as estimated by GENEEC2 is 16.07 μg/L.

RUN No. 1 FOR kasumin ON kiwifruit * INPUT VALUES *

--------------------------------------------------------------------

RATE (#/AC) No.APPS & SOIL SOLUBIL APPL TYPE NO-SPRAY INCORP

ONE(MULT) INTERVAL Koc (PPM ) (%DRIFT) (FT) (IN)

--------------------------------------------------------------------

0.089( 0.328) 4 10 138.0******* ORCHAR( 9.7) 0.0 0.0

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September 2013

FIELD AND STANDARD POND HALFLIFE VALUES (DAYS)

--------------------------------------------------------------------

METABOLIC DAYS UNTIL HYDROLYSIS PHOTOLYSIS METABOLIC COMBINED

(FIELD) RAIN/RUNOFF (POND) (POND-EFF) (POND) (POND)

--------------------------------------------------------------------

122.40 2 0.00 8.20- 1016.80 28.60 27.82

GENERIC EECs (IN MICROGRAMS/LITER (PPB)) Version 2.0 Aug 1, 2001

--------------------------------------------------------------------

PEAK MAX 4 DAY MAX 21 DAY MAX 60 DAY MAX 90 DAY

GEEC AVG GEEC AVG GEEC AVG GEEC AVG GEEC

--------------------------------------------------------------------

16.03 15.71 13.96 10.86 9.10

Calculation of acute risk quotients using GENEEC2 expected environmental concentrations

Table gives calculated acute risk quotients for each trophic level considering EEC estimated by GENEEC2

and lowest relevant toxicity figures.

Table 14: Acute risk quotients derived from the GENEEC2 model and toxicity data

Species

Peak EEC from

GENEEC2 (mg

a.i./L)

LC50 or EC50

(mg a.i./L) Acute RQ Presumption

Ground based application on kiwifruit

Fish

16.03 .10-3

2.4 0.007 Low risk

Crustacea 3.04 0.005 Low risk

Algae 2.64 0.006 Low risk

Conclusion for the acute aquatic risk assessment using GENEEC2 data

Formulation data have been used for the risk assessment. Low acute risk has been identified for the aquatic

organisms from the use of Kasumin in kiwifruits. However, it was impossible to evaluate the risk from the

significant metabolite (kasugamycinic acid) due to data lacking.

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September 2013

Calculation of chronic risk quotients using GEENEC2 expected environmental concentrations

Table gives calculated chronic risk quotients for each trophic level considering EEC estimated by GENEEC2

and lowest relevant toxicity figures.

Table 15: Chronic risk quotients derived from the GENEEC2 model and toxicity data

Species

Relevant EEC from

GENEEC2 (mg

a.i./L)*

NOEC

(mg

a.i./L)

Chronic RQ Presumption

Ground based application on kiwifruit

Fish (ELS, 32

days) 14.10

-3 9.5 0.001 Low risk

Crustacea (21

days) 14.10

-3 50 0.0003 Low risk

* EEC selected must be as close as possible from the exposure duration of the study selected for risk

assessment purpose.

Conclusion for the chronic aquatic risk assessment using GENEEC2 data

Low chronic risk has been identified for the aquatic organisms from the use of Kasumin in kiwifruit. However,

it was impossible to evaluate the risk from the significant metabolite (kasugamycinic acid) due to lack of data.

8.6. Sediment risk assessment

No data available so no risk assessment is possible.

8.7. Terrestrial risk assessment

For terrestrial organisms, Toxicity-Exposure Ratios (TERs) are used for earthworms and birds and Hazard

Quotient (HQ) are used for terrestrial invertebrates. This convention results in concern arising if a risk

quotient is less than the trigger value for earthworms and more than a trigger value for terrestrial

invertebrates. LOC developed by the European Union and adopted by the Staff allowing to determine

whether a substance poses an environmental risk are provided in the Table .

Table 16: Levels of concern as adopted by the Staff

Level of Concern (LOC) Presumption

Earthworm/ Birds

Acute TER < 10 High risk

Chronic TER < 5 High risk

Bees

HQ oral/contact > 50 High risk

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September 2013

Terrestrial invertebrates

HQ in-field/off-field ≥2 High risk

For more details about the different factors used for calculating TER and HQ refer to the relevant reference

documents listed in Table .

Earthworm risk assessment

Soil Predicted Environmental Concentration (PEC) determination

Both acute and reproductive earthworm tests are static tests where the test substance is applied to the

system only once at the beginning. Therefore the nominal dose levels in the test match initial concentrations

in the field and thus it is appropriate to use initial PEC values (no time-weighted averages) for the acute as

well as the long-term TER.

The concentration of active substance in the soil is calculated on the basis of the FOCUS (1997) document

‘Soil persistence models and EU registration’

( ) ( )

Soil concentrations of the active ingredient are calculated by assuming the deposition would mix into the top

5 cm of soil, and this soil would have a bulk density of 1,500 kg/m3, i.e. the deposition expressed in mg/m

2

would mix into 75 kg of soil.

In case of multiple applications, the following formula has to be used:

( )

( )

where:

n = number of applications

k = ln2/DT50 (day-1

)

i = interval between two consecutive applications (days)

DT50 = half life in soil (days) Use only DT50 values of lab test done at 10-20 oC and pH between 5 and 9. The

same DT50 as for GENEEC2 has been used (122.4).

e = 2.718 (constant)

PEC calculation results are summarized for each scenario in Table and Table .

Calculation of TERs

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September 2013

Table 17: Acute in-field TER value for earthworms

Scenarios PEC (mg/kg soil) LC50

(mg/kg soil) TER acute Presumption

Ground based

application on

kiwifruit

0.49 > 1000 > 2043 Low risk

Table 18: Acute off-field TER value for earthworms

Scenarios PEC (mg/kg soil) LC50

(mg/kg soil) TER acute Presumption

Ground based

application on

kiwifruit

0.05 > 1000 > 21057 Low risk

Conclusion for earthworm acute risk assessment

Low acute risks have been identified for the earthworm organisms from the use of Kasumin in kiwifruit, from

in-field as well as off-field exposures.

Conclusion for earthworm chronic risk assessment

No data are available about the chronic toxicity of kasugamycin on earthworms so no risk assessment was

performed.

Non-target plant risk assessment

Non target plants are defined as follows: Plants which are non-crop plants located outside the treatment

area.

The studies on vegetative vigour and seedling emergence of 10 species of non-target plants show that the

maximum application rate does not produce more than 50% of effects. Therefore no risk assessment is

necessary and the risk is considered to be low according to the Guidance Document on terrestrial

ecotoxicology (Table ). The Canadian authorities concluded that a buffer zone of 2 m was necessary to

protect the non-target plants however, their assessment is based on ER25 values, which is not consistent

with our current approach.

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September 2013

8.7.1. Bird risk assessment

No mortality was observed in the 3 acute tests and the 2 short-term tests on birds. In addition the NOEC

from the 2 chronic tests was the highest tested concentration (1000 ppm) so no risk assessment for birds

was performed due to the low toxicity of kasugamycin. The risks are considered to be very low.

8.7.2. Bee risk assessment

The risk to bees is assessed as follows:

( ) ( )

( ) ( )

Calculation of HQs

Table 19: Acute HQs values for bees

Species LD50 (µg a.s./bee) Application rate

(g a.i./ha)

Hazard

Quotient Presumption

Oral

Apis mellifera > 100 100 < 1 Low risk

Contact

Apis mellifera > 100 100 < 1 Low risk

Conclusions for bee risk assessment

The HQoral value is below the trigger value of 50, indicating that kasugamycin presents a low oral risk to

honeybees.

The HQcontact value is below the trigger value of 50, indicating that kasugamycin presents a low contact risk to

honeybees.

8.7.3. Non-target arthropod risk assessment

Note: Where limit tests are conducted, a low risk to non-target arthropods can be concluded when the effects

at the highest application rate multiply by MAF are below 50% (ESCORT2 workshop, 2000 – p12)

Calculation of HQs

( )

* application rate and LR50 must not differ in their units, i.e. must be related to either formulation or a.i. rates

137


September 2013

** Multiple application factor, refer to Appendix V, p 45 of ESCORT 2 Workshop, 2000. MAF = 1 when there is just one application.

(

)

* Overall 90th

percentile drift values are presented in Appendix VI , p 46 of ESCORT 2 Workshop, 2000.

** default value of 10

*** default value of 10

MAF (Multiple Application Factor) for 4 applications of kasugamycin is defined as 3.4 for soil organisms

(Pardosa spiders) and 2.7 for leave dwelling arthropods (Typhlodromus pyri).

A drift factor for kiwifruit of 23.61% (based on a minimum drift of 3 m and 4 applications) is used for the

off-field exposure calculations, based on recommendations made in ESCORT 2 and

SANCO/10329/2002 rev 2 final.

On this basis, the resultant in-field and off-field hazard quotients for Pardosa and Typhlodromus pyri are

shown in the Table and Table .

Table 20: In-field HQ values for Pardosa and Typhlodromus pyri

Species LR50

(g ai/ha)

Application rate

(g ai/ha) MAF

Hazard

Quotient Presumption

Pardosa > 180 100 3.4 1.89 Low risk

Typhlodromus pyri 102 100 2.7 2.65 High risk

Table 21: Off-field HQ values for Pardosa and Typhlodromus pyri

Sp

ecie

s

LR

50

(g a

i/h

a)

Ap

plicati

on

rate

(g a

i/h

a)

MA

F

Veg

eta

tio

n

facto

r

Co

rre

cti

on

facto

r

Dri

ft d

ista

nc

e

(m)/

% d

rift

Hazard

Qu

oti

en

t

Pre

su

mp

tio

n

Pardosa > 180 100 3.4 10 10 3/ 23.61 0.45 Low

risk

Typhlodromus

pyri 102 100 2.7 10 10 3/ 23.61 0.62

Low

risk

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September 2013

Conclusion for non-target arthropod risk assessments

Pardosa spiders are known to be less sensitive than the recommended indicator species (Typhlodromus pyri

and Aphidius rhopalosiphi). Therefore this species is not the most suitable for the risk assessment. Therefore

the conclusion of the risk assessment will be based on predatory mites only.

The in-field HQ value for kasugamycin is above the trigger value of 2 for Typhlodromus pyri, (the most

relevant species), indicating that this active substance is of high concern for non-target arthropods.

The off-field HQ value for kasugamycin is below the trigger value of 2, indicating that this active substance is

of low concern for non-target arthropods.

The study on Typhlodromus pyri shows that sub-lethal concentrations have a significant effect on the

reproduction. So even if the HQ is below the trigger for off-field situations on the basis of mortality, the off-

field population may still be at risk due to the effects on the reproduction.

No higher tier study is available to evaluate the potential of recovery and/or recolonisation.

Identification of persistent, bioaccumulative and toxic (PBT) and very

persistent and very bioaccumulative (vPvB) substances, components,

contaminants, or metabolites

Table 22: Screening criteria for Persistency, Bioaccumulation, and Toxicity

Type of data Criterion Screening assessment

Persistence

Ready biodegradability test Readily biodegradable Not P not vP

Enhanced ready biodegradability test Readily biodegradable Not P not vP

Specified tests on inherent biodegradability

Zahn-Wellens (OECD 302B)

MITI II test (OECD 302C)

≥ 70 % mineralisation (DOC

removal) within 7 d; log phase no

longer than 3d; removal before

degradation occurs below 15%;

no pre-adapted inoculums

≥ 70% mineralisation (O2 uptake)

within 14 days; log phase no

longer than 3d; no pre-adapted

inoculum

Not P

Not P

Biowin 2 (non-linear model prediction) and

Biowin 3 (ultimate biodegradation time)

or

Biowin 6 (MITI non-linear model prediction)

Does not biodegrade fast

(probability <0.5), and ultimate

biodegradation timeframe

prediction: ≥months (value < 2.2)

or

P

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September 2013

and Biowin 3 (ultimate biodegradation time)

Does not biodegrade fast

(probability <0.5) and ultimate

biodegradation timeframe

prediction: ≥months (value < 2.2)

P

Bioaccumulation

Convincing evidence that a substance can

biomagnify in the food chain (e.g. field data) e.g. BMF > 1

B or vB, definitive

assignment possible

Octanol-water partitioning coefficient

(experimentally determined or estimated by

QSAR)

Log Kow ≤ 4.5 Not B and not vB

Toxicity

Short-term aquatic toxicity EC50 or LC50 < 0.01 mg/L T, criterion considered to be

definitely fulfilled

Short-term aquatic toxicity EC50 or LC50 < 0.1 mg/L T

Avian toxicity (subchronic or chronic toxicity

or toxic for reproduction) NOEC < 30 mg/kg food T

The outcome of the assessment is summarized in the Table

Table 23: PBT/vPvB assessment for Kasugamycin

Yes No Cannot be determined

at this time

Does the substance or is

likely to contain PBT

components?

Component(s):

Remarks:

Are the metabolites

(mammalian and/or

environmental) PBT?

Metabolite(s):

Remarks:

Does the substance

contain or is likely to

contain vPvB (POP)

components?

Component(s):

Remarks:

Are the metabolites

(mammalian and/or

environmental) vPvB

(POP)?

Metabolite(s):

Remarks:

Does the substance

contain POP stipulated in

the Stockholm

Convention?

Substance:

Remarks:

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September 2013

Conclusion for PBT/vPvB assessment

Kasugamycin and Kasumin are not PBT/vPvB or do not contain PBT/vPvB substances.

Summary and conclusions of the ecological risk assessment

Low acute and chronic risks have been identified for the aquatic organisms from the use of Kasumin 2L on

kiwifruits. The same conclusion was drawn for acute risk on earthworms. The risk is also considered to be

low for the non-target plants, the bees and the birds from acute or chronic exposures.

Non-target arthropods are at a high risk in field and are likely to be at risk off-field due to the effects on

reproduction at sub-lethal doses.

It was not possible to assess all the risks due to substantial data gaps. The following studies would have

been necessary:

Effects of kasugamycinic acid on aquatic organisms (fish, invertebrates, algae).

Effects of kasugamycin on the reproduction of earthworms.

Effects of kasugamycin on the carbon transformation in soil.

Effects of kasugamycin on bee larvae.

Higher tier studies on non-target arthropods with kasugamycin.

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September 2013

Appendix C: Controls applying to Kasumin

Notes

The controls for these substances apply for the indefinite duration of the approval of the substances.

Please refer to the Hazardous Substances Regulations23 for the requirements prescribed for each control

and the modifications listed as set out in Section 7 of this document.

Hazardous Substances (Classes 6, 8, and 9 Controls) Regulations 2001

Code Regulation Description Variation

T1 11 – 27 Limiting exposure to toxic substances

through the setting of TELs

No TELs values are set for any

component of the substance at this

time; however, the following ADE and

PDE values have been set for

kasugamycin:

ADE = 0.113 mg/kg bw/day

PDEfood = 0.08 mg/kg bw/day

PDEdrinking water = 0.023 mg/kg bw/day

PDEother = 0.011 mg/kg bw/day

T2 29, 30 Controlling exposure in places of work

through the setting of WESs.

No WES values have been set for any

component of this substance at this

time.

T4 7 Requirements for equipment used to

handle substances

T5 8 Requirements for protective clothing

and equipment

T7 10

Restrictions on the carriage of toxic or

corrosive substances on passenger

service vehicles

Hazardous Substances (Identification) Regulations 2001


I1

6, 7, 32 –

35, 36(1) –

(7)

Identification requirements, duties of

persons in charge, accessibility,

comprehensibility, clarity and

durability

I8 14 Priority identifiers for toxic

substances

I9 18 Secondary identifiers for all

hazardous substances

23

The regulations can be found on the New Zealand Legislation website; http://www.legislation.co.nz

http://www.legislation.co.nz/

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I16 25 Secondary identifiers for toxic

substances

The concentration cut-offs that

trigger the requirement for labelling

of components are set out in the

following table:

HSNO Classification

Cut-off for label % (I16)

6.5A, 6.5B, 6.6A, 6.7A

0.1

6.6B 1

6.7B 1

6.8A, 6.8C 0.3

6.8B 3

6.9A, 6.9B 10

I17 26 Use of generic names

I18 27 Requirements for using concentration

ranges

I19 29 – 31

Additional information requirements,

including situations where

substances are in multiple packaging

I21 37 – 39, 47

– 50 General Documentation requirements

I28 46 Specific Documentation requirements

for toxic substances

I30 53 Advertising corrosive and toxic

substances

Hazardous Substances (Packaging) Regulations 2001


P1 5, 6, 7(1), 8 General packaging requirements

P3 9

Criteria that allow substances to be

packaged to a standard not meeting

Packing Group I, II or III criteria

P13 19 Packaging requirements for toxic

substances

PS4 Schedule 4 Packaging requirements as specified

in Schedule 4

Hazardous Substances (Disposal) Regulations 2001


D4 8 Disposal requirements for toxic and

corrosive substances

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September 2013

D6 10 Disposal requirements for packages

D7 11, 12

Information requirements for

manufacturers, importers and

suppliers, and persons in charge

D8 13, 14

Documentation requirements for

manufacturers, importers and

suppliers, and persons in charge

Hazardous Substances (Emergency Management) Regulations 2001


EM1 6, 7, 9 – 11 Level 1 information requirements for

suppliers and persons in charge

EM6 8(e) Information requirements for toxic

substances

EM8 12 – 16, 18

– 20

Level 2 information requirements for

suppliers and persons in charge

Hazardous Substances (Tank Wagon and Transportable Containers) Regulations 2004

Code Regulation Description

Tank

Wagon

4 to 43 as

applicable Controls relating to tank wagons and transportable containers.

Schedule 8 of the Hazardous Substances (Dangerous Goods and Scheduled Toxic Substances) Transfer

Notice 2004


Sch 8 Schedule 8 This schedule prescribes the controls for

stationary container systems. The

requirements of this schedule are detailed

in the consolidated version of the

Hazardous Substances (Dangerous

Goods and Schedule Toxic Substances)

Transfer Notice 2004, available from

http://www.epa.govt.nz/Publications/Trans

fer-Notice-35-2004.pdf

Additional controls


Water 77A This substance must not be applied onto, over or into water24

App Rate 77A Kasumin may be applied at a maximum application rate of 100 g

24 Where "water" means water in all its physical forms, whether flowing or not, and whether over or under ground, but does not include water in any form while in a pipe, tank or cistern or water used in the dilution of the substance prior to application or water used in the dilution of the substance prior to application or water used to rinse the container after use

http://www.epa.govt.nz/Publications/Transfer-Notice-35-2004.pdf

http://www.epa.govt.nz/Publications/Transfer-Notice-35-2004.pdf

144


September 2013


kasugamycin/ha per application, and up to four applications of Kasumin

per year, with a minimum interval of 10 days between applications.

Ground

Bas

77A Kasumin must only be applied via ground-based methods25

R-12 77A Restricted Entry Interval (REI)

(1) The REI for this substance is 40 days.

(2) The person in charge of the application area shall ensure that no

person who is authorised to be there enters the application area until

the end of the REI.

(3) Despite (2), a person may enter the application area before the end

of the REI—

(a) if PPE and RPE is worn as if that person is applying the

substance; and

(b) if entering an indoor treated area, for the purpose of carrying

out tasks associated with ventilation of the building or structure.

LABEL STATEMENT

(4) A person must not supply a hazardous substance to any other

person unless the substance label shall show the requirements for the

REIs and corresponding PPE, in accordance with (1) to (3).

(5) A person who is in charge of a hazardous substance must ensure

that the substance label shows the information required by (4).

R-13 77A Approved handler requirements

(1) The requirements of regulation 9 of Hazardous Substances

(Classes 6, 8, and 9 Controls) Regulations 2001, and regulations 4 to 6

of Hazardous Substances (Personnel Qualifications) Regulations 2001

apply to this substance.

LABEL STATEMENT


person unless the substance label specifies that the substance must

only be applied by an approved handler, or under the direct supervision

of an approved handler.



This regulation is inserted immediately after regulation 9:

9A Exception to approved handler requirement for transportation

of packaged substances

(1) Regulation 9 is deemed to be complied with if—

(a) in the case of a hazardous substance being transported on land,—

(i) in the case of a substance being transported by rail, the person who

drives the rail vehicle that is transporting the substance is fully trained

25 Ground-based methods of applying pesticides include, but are not limited to, application by ground boom, airblast or knapsack, and do not include aerial application methods.

145


September 2013


in accordance with the approved safety system for the time being

approved under section 6D of the Transport Services Licensing Act

1989; and

(ii) in every other case, the person who drives, loads, and unloads the

vehicle that is transporting the substance has a current dangerous

goods endorsement on his or her driver licence; and

(iii) in all cases, Land Transport Rule: Dangerous Goods 1999 (Rule

45001) is complied with; or

(b) in the case of a hazardous substance being transported by sea, one

of the following is complied with:

(i) Maritime Rules: Part 24A – Carriage of Cargoes – Dangerous Goods

(MR024A):

(ii) International Maritime Dangerous Goods Code; or

(c) in the case of a hazardous substance being transported by air, Part

92 of the Civil Aviation Rules is complied with.

(2) Subclause (1)(a)—

(a) does not apply to a tank wagon or a transportable container to

which the Hazardous Substances (Tank Wagons and Transportable

Containers) Regulations 2004 applies; but

(b) despite paragraph (a), does apply to an intermediate bulk container

that complies with chapter 6.5 of the UN Model Regulations.

(3) Subclause (1)(c)—

(a) applies to pilots, aircrew, and airline ground personnel loading and

managing hazardous substances within an aerodrome; but

(b) does not apply to the handling of a hazardous substance in any

place that is not within an aerodrome.

(4) In this regulation, UN Model Regulations means the 17th revised

edition of the Recommendation on the Transport of Dangerous Goods

Model Regulations, published in 2011 by the United Nations.

R-4 77A Spray drift mitigation

(1) No person may apply the substance in a manner that results in

adverse effects beyond the boundary of the subject property.

(2) A person applying the substance must take all practicable steps to

avoid off-target movement of the substance.

LABEL STATEMENT

(3) The following statement must appear on the substance label:

The person applying this substance must not cause adverse effects

beyond the boundary of the treated property, and must also avoid

adverse effects from spray drift occurring. Mitigation measures

employed must be recorded as part of the application records.


person unless the substance label includes the statement specified in

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(3).



The following subclause is added to regulation 6(1) of the Hazardous

Substances (Classes 6, 8, and 9 Controls) Regulations 2001, after

subclause (g):

(h) details of measures taken to ensure that there are no adverse

effects beyond the boundary of the subject property into an adjoining

property or sensitive area.

Label 77A The label must include the following statement:

Kasumin contains the antibiotic, kasugamycin, and may be harmful to

beneficial organisms, such as predatory mites, earthworms and bees

and other pollinators.

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Appendix D: References

AgDRIFT

Crocker D., Hart A., Gurney J. and McCoy C. (2002). Project PN0908: Methods for estimating daily food

intake of wild birds and mammals. York: Central Science Laboratory.

EC (2002). SANCO/10329/2002 rev 2 final. Guidance document on terrestrial ecotoxicology under Council

Directive 91/414/EEC, 17 October 2002.

EC (2002). SANCO/4145/2000 final. Guidance document on risk assessment for birds and mammals under

Council Directive 91/414/EEC, 25 September 2002.

EC (2003). Technical guidance document on risk assessment in support of Commission Directive 93/67/EEC

on Risk Assessment for new notified substances, Commission Regulation (EC) No 1488/94 on Risk

Assessment for existing substances, Directive 98/8/EC of the European Parliament and of the Council

concerning the placing of biocidal products on the market – Part II.

ECHA (2008). Guidance on information requirements and chemical safety assessment, Chapter R.10:

Characterisation of dose [concentration]-response for environment.

ECHA (2008). Guidance on information requirements and chemical safety assessment, Chapter R.11: PBT

assessment.

ECHA (2010). Guidance on information requirements and chemical safety assessment, Chapter R.16:

Environmental Exposure Estimation.

EFSA (2009). Guidance of EFSA. Risk assessment to birds and mammals, 17 December 2009.

EFSA (2009). Calculator tool. www.efsa.europa.eu/en/efsajournal/pub/1438.htm 2009.

ESCORT 2 (2000). Guidance document on regulatory testing and risk assessment procedures for plant

protection products with non-target arthropods. From ESCORT 2 Workshop, 21/23 March 2000.

FOCUS (1997). Soil persistence models and EU registration. The final report of the work of the Soil

Modelling Work group of FOCUS (FOrum for the Co-ordination of pesticide fate models and their USe), 29

February 1997.

Ganzelmeier H. D., Rautmann R., Spangenberg M., Streloke M., Herrmann H.-J., Wenzelburger and H.-F.

Walter (1995). Studies on the spray drift of plant protection products. Heft 305, Blackwell Wissenschafts-

Verlag GmbH, Berlin: 111 ppJager T. (1998). Mechanistic approach for estimating bioconcentration of

organic chemicals in earthworms (Oligochaeta). Environ Toxicol Chem, 17, 2080-2090.

Smit C.E. (2005). Energy and moisture content and assimilation efficiency of bird and mammal food.

Rautmann D., Streloke M. and R. Winkler (2001). New basic drift values in the authorization procedure for

plant protection products. Mitt. Biol. Bundesanst. Land- Forstwirtsch. 383:133-141.

RIVM report 601516013, 57–71.

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Urban D. J. and Cook N. J. (1986). Hazard Evaluation Division Standard Evaluation Procedure: Ecological

Risk Assessment. EPA 540/9-85-001. United States Environmental Protection Agency Office of Pesticide

Programs, Washington DC, USA.

US EPA (2002). (GEN)eric (E)stimated (E)nvironmental (C)oncentration Model Version 2.0, 01 August 2002.

US EPA (2004). Overview of the Ecological Risk Assessment Process in the Office of Pesticide Programs,

U.S. Environmental Protection Agency. Endangered and threatened Species Effects Determinations, 23

January 2004.

Veith G.D., Defoe D.L. and Bergstedt B.V. (1979). Measuring and estimating the bioconcentration factor of

chemicals on fish. J. Fish. Res. Board Can., 36, 1040-1048.

epa staff evaluation and review report

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