No 528/2012 Concerning the Making Available on the Market and Use of Biocidal Products

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Regulation (EU) No 528/2012 concerning the making available on the market and use of biocidal products

DRAFT RISK ASSESSMENT OF A BIOCIDAL PRODUCT FOR NATIONAL AUTHORISATION APPLICATIONS (submitted by the eCA)

® BIOPREN 50 LFL Fly Larvicide Concentrate

Product type(s) 18

S-methoprene as included in the Union list of approved active substances

Asset Number NL-0018671-0000

Evaluating Competent Authority: The Netherlands

Date: 04/06/2021 NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Table of Contents

1 CONCLUSION...... 4 2 ASSESSMENT REPORT ...... 6

2.1 SUMMARY OF THE PRODUCT ASSESSMENT ...... 6 2.1.1 Administrative information...... 6 2.1.1.1 Identifier of the product ...... 6 2.1.1.2 Authorisation holder ...... 6 2.1.1.3 Manufacturer(s) of the products ...... 6 2.1.1.4 Manufacturer(s) of the active substance(s) ...... 6 2.1.2 Product composition and formulation ...... 7 2.1.2.1 Identity of the active substance ...... 7 2.1.2.2 Candidate(s) for substitution ...... 7 2.1.2.3 Qualitative and quantitative information on the composition of the biocidal product ...... 8 2.1.2.4 Qualitative and quantitative information on the composition of the biocidal product family 2 ...... 8 2.1.2.5 Information on technical equivalence ...... 8 2.1.2.6 Information on the substance(s) of concern ...... 8 2.1.2.7 Type of formulation ...... 9 2.1.3 Hazard and precautionary statements ...... 9 2.1.4 Authorised use(s) ...... 9 2.1.4.1 Use description ...... 10 2.1.4.2 Use-specific instructions for use ...... 11 2.1.4.3 Use-specific risk mitigation measures ...... 11 2.1.4.4 Where specific to the use, the particulars of likely direct or indirect effects, first aid instructions and emergency measures to protect the environment...... 11 2.1.4.5 Where specific to the use, the instructions for safe disposal of the product and its packaging ...... 11 2.1.4.6 Where specific to the use, the conditions of storage and shelf-life of the product under normal conditions of storage 11 2.1.5 General directions for use ...... 12 2.1.5.1 Instructions for use ...... 12 2.1.5.2 Particulars of likely direct or indirect effects, first aid instructions and emergency measures to protect the environment ...... 13 2.1.5.3 Instructions for safe disposal of the product and its packaging ...... 13 2.1.5.4 Conditions of storage and shelf-life of the product under normal conditions of storage ...... 13 2.1.6 Other information ...... 13 2.1.7 Packaging of the biocidal product ...... 14 2.1.8 Documentation ...... 14 2.1.8.1 Data submitted in relation to product application ...... 14 2.1.8.2 Access to documentation ...... 14 2.2 ASSESSMENT OF THE BIOCIDAL PRODUCT ...... 15 2.2.1 Intended use(s) as applied for by the applicant ...... 15 2.2.2 Physical, chemical and technical properties ...... 15 2.2.3 Physical hazards and respective characteristics ...... 30 2.2.4 Methods for detection and identification ...... 36 2.2.5 Efficacy against target organisms ...... 39 2.2.5.1 Function and field of use ...... 39 2.2.5.2 Organisms to be controlled and products, organisms or objects to be protected ...... 39 2.2.5.3 Effects on target organisms, including unacceptable suffering ...... 39 2.2.5.4 Mode of action, including time delay ...... 39 2.2.5.5 Efficacy data ...... 40 2.2.5.6 Occurrence of resistance and resistance management ...... 45 2.2.5.7 Known limitations ...... 46 2.2.5.8 2.2.5.8 Evaluation of the label claims ...... 46 2.2.5.9 Relevant information if the product is intended to be authorised for use with other biocidal product(s)... 50 2.2.6 Risk assessment for human health ...... 50 2.2.6.1 Assessment of effects on Human Health ...... 50

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Others ...... 60 2.2.6.2 Exposure assessment ...... 60 2.2.6.3 Risk characterisation for human health ...... 91 2.2.7 Risk assessment for animal health...... 98 2.2.8 Risk assessment for the environment ...... 99 2.2.8.1 Effects assessment on the environment ...... 100 2.2.8.2 Exposure assessment ...... 111 2.2.8.3 Risk characterisation ...... 121 2.2.9 Measures to protect man, animals and the environment ...... 128 2.2.10 Assessment of a combination of biocidal products ...... 130 2.2.11 Comparative assessment ...... 131 3 ANNEXES ...... 132

3.1 LIST OF STUDIES FOR THE BIOCIDAL PRODUCT ...... 132 3.2 OUTPUT TABLES FROM EXPOSURE ASSESSMENT TOOLS ...... 137 3.3 NEW INFORMATION ON THE ACTIVE SUBSTANCE ...... 139 3.4 RESIDUE BEHAVIOUR ...... 140 3.5 SUMMARIES OF THE EFFICACY STUDIES (B.5.10.1-XX ) ...... 140 3.6 CONFIDENTIAL ANNEX ...... 140 3.7 ENVIRONMENTAL RISK ASSESSMENT ...... 140 3.8 OTHER ...... 141

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

1 CONCLUSION

BIOPREN® 50 LFL Fly Larvicide Concentrate is a partly microencapsulated viscous turquoise and slightly sweetish liquid formulation containing as active substance 5.00 % S- methoprene.

Based on the efficacy studies it can be concluded that the product is sufficiently efficacious against larvae of Musca domestica , Stomoxys calcitrans and Eristalis tenax at an application rate of 2.3-2.4 g/m 2. Dosage to be applied to treat 100 m 2 surface: 230-240 ml of BIOPREN 50 LFL concentrate diluted to 5 litres with water. Maximum sprayer flow rate: 0.3-0.5 L/min. Efficacy has been demonstrated up to 12 weeks after treatment in cowsheds, pigsties and sheepfolds Expected time delay from the beginning of the application to manifestation of the biocidal effect is 2-4 weeks.

Based on the physical, chemical and technical characteristics the eCA considers the product as sufficiently described to fulfil the conditions for granting an authorization.

The applicant has requested read-across to be used for several test studies in order to support the shelf life of the product. Based on the combined results obtained for the product BIOPREN® 50 LFL Fly Larvicide Concentrate and BIOPREN® 50 LFL mosquito Larvicide Concentrate a shelf life of 30 months in COEX PE/PA and HDPE is supported.

The analytical method used to determine the content of active substance has been sufficiently validated.

The product is not classified based on its physical hazards and characteristics.

Regarding human health, Biopren 50 LFL Fly larvicide concentrate has shown acceptable risks for all presented scenarios when considered separately, except for the application by spraying without protective equipment. Therefore, PPE is required for this scenario (gloves and coverall). When PPE is worn, the risk is acceptable for all scenarios, based on all models. BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE is classified with : H317: May cause an allergic skin reaction. Furthermore, the diluted product is classified with EUH208: Contains Polyethylene glycol (15)-hydroxystearate. May produce an allergic reaction. Based on this classification with H317 and taking into account the product is applied by spraying, P280: Wear protective gloves and protective clothing is prescribed. The risk of combined scenarios is acceptable when professional users wear PPE. Furthermore, the following general risk mitigation measures are included in the SPC: ‘Avoid contact with skin and eyes’ and ‘Wash hands thoroughly with hot water and soap after use’. No additional risk mitigation measures are considered needed.

The general public is only indirectly exposed to the diluted product. No adverse systemic health effect is expected for the general public by use of BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE. Athough BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE is classified with H317: May cause an allergic skin reaction, the diluted product would be classified with EUH208: Contains Polyethylene glycol (15)-hydroxystearate. May produce an allergic reaction. . However, the diluted product is applied to manure. Furthermore, considering the EUH208 classification, this communicates that this effect could occur in already sensitized persons. A child may be exposed by dermal contact after accidentally touching the treated manure, and therefore, no local effects are expected for the general public.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

The dietary exposure assessment concluded no concern based on the theoretical maximum consumption of 0.0087 mg/kg bw/d using the food basket, which is 4.0% of the derived ADI. According to BPR guidance, no MRL needs to be derived if the dietary exposure is below 30% of the ADI.

Regarding the environment the product is classified as H411. The precautionary statements P391 and P501 are added. Emission of residues of the product to the sewer results in unacceptable risks for the aquatic environment, but emission to the manure is acceptable. Therefore a risk mitigation measure stating that residues of the product must be disposed to the manure storage and not to the sewer is added. Discharge of residues to the manure does not results in unacceptable risks for the aquatic and terrestrial environment.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

2 ASSESSMENT REPORT

2.1 SUMMARY OF THE PRODUCT ASSESSMENT 2.1.1 Administrative information

2.1.1.1 Identifier of the product

Identifier 1 Country (if relevant) NL BIOPREN ® 50 LFL Fly Larvicide Concentrate

2.1.1.2 Authorisation holder

Name and address of the Name Babolna Bio Ltd authorisation holder Address Szállás utca 6, Budapest, H -1107 Authorisation number NL-0018671-0000 Date of the authorisation 4 June 2021 Expiry date of the 4 June 2031 authorisation

2.1.1.3 Manufacturer(s) of the products

Name of manufacturer Babolna Bio Ltd Address of manufacturer Szállás utca 6, Budapest, H -1107 Location of manufacturing Dr Köves János út 1-3, Babolna, H-2943 sites

2.1.1.4 Manufacturer(s) of the active substance(s)

Active substance S-methoprene Name of manufacturer Babolna Bio Ltd Address of manufacturer Szállás utca 6, Budapest, H -1107 Location of manufacturing Szállás utca 6, Budapest, H-1107 sites

1 Please fill in here the identifying product name from R4BP.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

2.1.2 Product composition and formulation

NB: the full composition of the product according to Annex III Title 1 should be provided in the confidential annex.

Does the product have the same identity and composition as the product evaluated in connection with the approval for listing of the active substance(s) on the Union list of approved active substances under Regulation No. 528/2012? Yes No x

2.1.2.1 Identity of the active substance Main constituent(s) ISO name S-methoprene IUPAC or EC name Isopropyl-(2E,4E, 7S)-11-methoxy-3,7,11-trimethyl-2,4- dodecadienoate EC number Not available CAS number 65733 -16 -6 Index number in Not available Annex VI of CLP Minimum purity / 95% w/w content Structural formula CH3

CH3O C O CH

O CH3

2.1.2.2 Candidate(s) for substitution S-methoprene is not a candidate for substitution.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

2.1.2.3 Qualitative and quantitative information on the composition of the biocidal product 2

Common name IUPAC Function CAS EC number Content (%) name number S-methoprene Isopropyl- Active 65733-16-6 Not 5.26 (pure: (2E,4E, 7S)- substance available free 1.25, 11- micro-encaps.d methoxy- 3.75 total: 3,7,11- 5.00) trimethyl- 2,4- dodecadieno ate Polyethylene glycol Octadecanoi Non-active 70142-34-6 Not 11.58 (15)-hydroxystearate c acid, 12- substance available hydroxy-, polymer with .alpha.- hydro- .omega.- hydroxypoly (oxy-1,2- ethanediyl) The composition details of the full formulation are contained within the confidential annex 3.6.1. eCA remark: In the non-confidential part a range is considered acceptable for non-active substances in order to avoid including the exact amount in the non-confidential part.

2.1.2.4 Qualitative and quantitative information on the composition of the biocidal product family 2

Not relevant

2.1.2.5 Information on technical equivalence The notified source of S-methoprene is the same as that considered for Annex I inclusion under Council Directive 98/8/EC. The applicant Babolna Bio Ltd. is the Annex I notified source of S-methoprene active substance.

2.1.2.6 Information on the substance(s) of concern Please see the confidential annex for further details.

The only substance of concern in the product is polyethylene-glycol-15-hydroxy-stearate due to its hazard for human toxicity.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

2.1.2.7 Type of formulation

ZW (CS+EW)

2.1.3 Hazard and precautionary statements 3

Classification and labelling of the products according to the Regulation (EC) 1272/2008 [It should also be stated if some P statements triggered by the criteria in CLP has been excluded due to the risk assessment.]

Classification Hazard category Skin Sens 1. Aquatic Chronic 2 Hazard statement H317: May cause an allergic skin reaction. H411: Toxic to aquatic life with long lasting effects.

Labelling Signal words Warning Hazard statements H317: May cause an allergic skin reaction. H411: Toxic to aquatic life with long lasting effects. Precautionary P261: Avoid breathing spray. statements P280: Wear protective gloves/protective clothing. P302+P352: IF ON SKIN: Wash with plenty of water. P333+P313: If skin irritation or rash occurs: Get medical advice/attention. P363: Wash contaminated clothing before reuse. P273: Avoid release to the environment P391: Collect spillage. P501: Dispose of contents/container to… [… in accordance with local/regional/national/international regulation]

Note Based on the presence of Polyethylene-glycol-15-hydroxy- stearat the product needs classiciation with H317. In accordance to CLP, Polyethylene-glycol-15-hydroxy-stearat needs to be declared on the label.

2.1.4 Authorised use(s)

3 For micro-organisms based products: indication on the need for the biocidal product to carry the biohazard sign specified in Annex II to Directive 2000/54/EC (Biological Agents at Work).

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

2.1.4.1 Use description 4 Table 1. Use # 1 – Insecticide – Professional use - larvicide

Product Type PT18 - Insecticides, acaricides and products to control other arthropods (Pest control) Where relevant, an Insecticide exact description of the authorised use Target organism (including Musca domestica – House fly - larvae development stage) Stomoxys calcitrans – Stable fly - larvae

Eristalis tenax – Dronefly - larvae Field of use Indoor, Outdoor; To be used in closed livestock buildings (piggeries, sheepfolds and cattle farms). Outdoor use only concerns the treatment of manure/slurry kept in a concrete basin (leak- proof insulated). Application method(s) Spraying Application rate(s) and Slatted floor: Apply on the 3 rd day after the introduction of frequency new livestock. Treat the whole floor of manure-pit. It is necessary to repeat treatment after each removal of dung.

Deep litter: Treat the litter before the introduction of new livestock. Repeat the treatment by each manure layer of 8 - 10 cm thick.

Dosage to be applied to treat 100 m 2 surface: 230-240 ml of BIOPREN 50 LFL concentrate diluted to 5 litres with water. Note: apply this dosage on approx. 10 cm thick manure Application rate: 2.3-2.4 g/m 2. Maximum sprayer flow rate: 0.3-0.5 L/min Maximum number of applications : six per year

The product can control the number of newly hatched adult flies up to 12 weeks after treatment. Expected time delay from the beginning of the application to the beginning of decrease in number of flies emerging is 2 -4 weeks. The biocidal effect is fully demonstrated after 8-12 weeks. Category(ies) of users Professional, trained professional Pack sizes and Bottle: COEX PE/PA or HDPE, up to 10 L packaging material Drum: HDPE drum up to 55 L Can: COEX PE/PA or HDPE, up to 10 L

4 Copy this section as many times as necessary (one table per use, together with any instructions for use, risk mitigation measures and other directions for use that are use-specific. It has to be noted that in accordance with Document CA-May14-Doc.5.6 – Final, the SPC of a biocidal product presents the authorised uses as a number of pre-defined uses to which the product label shall have full correspondence.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

2.1.4.2 Use-specific instructions for use 5

See general directions for use

2.1.4.3 Use-specific risk mitigation measures

See general directions for use

2.1.4.4 Where specific to the use, the particulars of likely direct or indirect effects, first aid instructions and emergency measures to protect the environment

See general directions for use

2.1.4.5 Where specific to the use, the instructions for safe disposal of the product and its packaging

See general directions for use

2.1.4.6 Where specific to the use, the conditions of storage and shelf-life of the product under normal conditions of storage

See general directions for use

5 Describe the necessary instructions for use like for example: period of time needed for the biocidal effect; the interval to be observed between applications of the biocidal product or between application and the next use of the product treated, or the next access by humans or animals to the area where the biocidal product has been used, including particulars concerning decontamination means and measures and duration of necessary ventilation of treated areas; particulars for adequate cleaning of equipment; particulars concerning precautionary measures during transport; precautions to be taken to avoid the development of resistance.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

2.1.5 General directions for use

2.1.5.1 Instructions for use 6

Shake the concentrate well before use. Once the required quantity of concentrate is poured, close the bottle immediately.

Fill the sprayer with water to half of its volume, measure the quantity of concentrate given in the table and pour into the sprayer . After that, fill the sprayer to the desired volume, then mix the solution thoroughly. Keep the prepared working solution closed and use it up within 24 hours after preparation. Shake the working solution from time to time during treatment or every time f ollowing longer standing.

Rinse the empty container three times and add rinses to the spray tank.

Contact of livestock with the product should be avoided. If this is not feasible, remove the animals during treatment. Treat the manure accumulated under t he slatted floor and the cage or the deep litter system. Do not spray the working solution directly next to the feeding or drinking place or places where the animals may consume the product. It is important to spray the surface of the manure within 3 days after the introduction of livestock, then treat every newly formed manure layer of 10 cm thick. After each removal of the manure, evenly treat the surface of the manure remaining in the stable.

In the case of very dry substrates effect of the product may develop slower. Adding 100-200 mL water/m2 to the substrate may facilitate development of larvicide effect in the case of very dry environmental conditions.

Slatted floor: Apply on the 3 rd day after the introduction of new livestock. Treat the whole floor of manure-pit. It is necessary to repeat treatment after each removal of dung.

Deep litter: Treat the litter before the introduction of new livestock. Repeat the treatment by each manure layer of 8 - 10 cm thick.

Dosage to be applied to treat 100 m2 surface: 230-240 ml of BIOPREN 50 LFL concentrate diluted to 5 litres with water. Note: apply this dosage on approx. 10 cm thick manure

Application rate: 2.3-2.4 g/m 2. Maximum sprayer flow rate: 0.3-0.5 L/min

-Inform the registration holder if the treatment is ineffective. -Take into account the life cycle and characteristics of target insects to adapt treatments. In particular, target the most susceptible stage of the pest, timing of applications and areas to be treated. -Do not apply the product i n areas where resistance to the active substance (s) contained in this product is supected or established

6 Describe the necessary instructions for use like for example: period of time needed for the biocidal effect; the interval to be observed between applications of the biocidal product or between application and the next use of the product treated, or the next access by humans or animals to the area where the biocidal product has been used, including particulars concerning decontamination means and measures and duration of necessary ventilation of treated areas; particulars for adequate cleaning of equipment; particulars concerning precautionary measures during transport; precautions to be taken to avoid the development of resistance.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Risk mitigation measures

Wear appropriate personal protection during mixing, loading and application of the product, including cleaning of the spray equipment (gloves and coverall).

To avoid any risk to humans and the environment, observe the directions for use and the provisions concerning health at work. Do not eat or smoke during work. Avoid contact with skin and eyes. Wash hands thoroughly with hot water and soap after use.

It is forbidden to use BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE in case effluent/wastewater of the stable and/or manure deposit can directly leach into the sewer.

2.1.5.2 Particulars of likely direct or indirect effects, first aid instructions and emergency measures to protect the environment

Likely direct or indirect adverse effects: • Repeated exposure may cause allergic disorders.

First aid measures: • IF IN EYES: Rinse cautiously with water for 15 minutes. Remove contact lenses, if present and easy to do. • IF ON SKIN (or hair): Take off immediately all contaminated clothing. Rinse skin with water/shower. • IF exposed or concerned: Get medical advice/attention.

2.1.5.3 Instructions for safe disposal of the product and its packaging

The remnants of the product should be disposed of as hazardous waste.

2.1.5.4 Conditions of storage and shelf-life of the product under normal conditions of storage

Store in the closed, original container in a dry, cool, well ventilated area out of direct sunlight. Store product at temperatures between 5 and 40 °C.

Shelf life: 30 months.

2.1.6 Other information

To prevent the development of insecticide resistance it is advised to: - Products should always be used in accordance with label instructions. - It is advised to use insecticides with different mode of action in rotation during the pest control program, especially if an extended period of control is required. - Where possible, chemical treatments should be recommended to be combined with application of non-chemical measures (e.g. sticky traps or fly traps with UV lamps). - Sufficient elimination of pest insects should be attempted in infested areas. - Levels of effectiveness should be monitored, and instances of reduced effectiveness should be investigated for possible evidence of resistance in the frame of IPM programs.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

2.1.7 Packaging of the biocidal product

Type of Size/volume Material Type and Intended Compatibility packaging of the of the material user (e.g. of the packaging packaging of professional, product with closure(s) non- the proposed professional) packaging materials (Yes/No) bottle up to 10 L opaque HDPE with professional yes white HIS liner COEX airtight PE/PA or closure cap HDPE can up to 10 L opaque HDPE with professional yes white HIS liner COEX airtight PE/PA or closure cap HDPE drum up to 55 L opaque HDPE professional yes white HDPE closure cap eCA remark: The information provided on the packaging of the biocidal product is considered sufficient.

2.1.8 Documentation

2.1.8.1 Data submitted in relation to product application Please find the list of studies under 3.3 New information on the active substance point.

2.1.8.2 Access to documentation Babolna Bio Ltd is the owner of S-methoprene active substance dossier and also the sponsor of the studies prepared with the Biopren 50 LFL Fly Larvicide Concentrate.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

2.2 ASSESSMENT OF THE BIOCIDAL PRODUCT 2.2.1 Intended use(s) as applied for by the applicant

The uses below are the ones applied for by the applicant, without any changes by the e- CA. These uses are assessed in the following chapters. See 2.1.4 for the authorised uses, after assessment of the dossier.

Table 2. Intended use # 1 – PCO 7

Product Type(s) PT18 - Insecticides, acaricides and products to control other arthropods (Pest control) Where relevant, an exact The product is based on S-methoprene active substance which description of the is a juvenile hormone analogue that inhibits the development authorised use of pupae into imagoes. It does not kill adult flies; As a result of its special liquid formulation, the product has both immediate and residual action, and – depending on the conditions – preserves its efficacy for as long as 12 weeks. Target organism (including development Muscidae: house fly larvae (Musca Domestica) stage) Muscidae: stable flies larvae (Stomoxys calcitrans)

Musciade: rat-tailed maggot larvae (Eristalis tenax) Field of use In closed piggeries, cattle and poultry farms, as well as in horse, goat and sheep stables and other livestock buildings, to prevent fly larvae breeding the manure from developing into imagoes. Application method(s) spraying Application rate(s) and Depends on the rate of infestation. The product can control frequency the number of newly hatched adult flies for a period of 12 weeks after treatment. Category(ies) of user(s) Professionals Pack sizes and packaging Please see the relevant section. material

2.2.2 Physical, chemical and technical properties

Babolna Bio Ltd, the applicant proposes read-across to the relevant GLP study of Biopren 50 LML mosquito larvicide concentrate, which was first authorised in Italy in 2019 and subsequently in 5 other EU countries. This product is exactly identical to Biopren 50 LFL fly larvicide concentrate, except for the colour, which is different to distinguish between the two products. This minor difference in the colour is not expected to have any influence on the physico-chemical properties of the product. Therefore, tests performed with Biopren 50 LML mosquito larvicide concentrate are considered relevant for this current product as well. The difference between the two product is the following: • Biopren 50 LFL fly larvicide concentrate: blue colour

7 Copy this section as many times as necessary (one table per use).

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

• Biopren 50 LML mosquito larvicide concentrate: no colouring agent present, the difference is compensated with water The composition of the product Biopren 50 LML mosquito larvicide concentrate can be found in the confidential annex: COMPARISION TABLE OF BIOPREN 50 LFL AND BIOPREN 50 LML COMPOSITION FOR SUPPORTING READ-ACROSS DATA

Purity of Guideline the test Property Results Reference and Method substance (% (w/w) Physical state at 20 °C OPPTS 4.95% Viscous liquid xxxx,Study and 101.3 kPa 830.6303 Batch No.: Report: Physical FN/319 Determination State, US of the EPA, 1996 Appearance of BIOPREN 50 LFL fly larvicide concentrate, Study No.: 484-630-0532 Colour at 20 °C and 101.3 OPPTS 4.95% Turquoise xxxx Study kPa 830.6302 Batch No.: Report: Color, US FN/319 Determination EPA, 1996 of the Appearance of BIOPREN 50 LFL fly larvicide concentrate, Study No.: 484-630-0532 Odour at 20 °C and 101.3 OPPTS 4.95% Slightly xxxx Study kPa 830.6304 Batch No.: sweetish Report: Odor, US FN/319 Determination EPA, 1996 of the Appearance of BIOPREN 50 LFL fly larvicide concentrate, Study No.: 484-630-0532 Acidity / alkalinity CIPAC MT 4.89% pH neat = 7.0 at xxxx Study 75.3, Batch No.: 20±1°C (n=3) Report: Determinatio 136182 Determination n of pH of the pH Values OECD pH 1% w/v aqueous Values of No. 122, dilution = 6.5 at Biopren 50 Determinatio 20±1°C (n=3) LML mosquito n of pH, larvicide Acidity and concentrate, Alkalinity, Study No.: 484-122-4159

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Purity of Guideline the test Property Results Reference and Method substance (% (w/w) adopted July 26, 2013 pH neat = 6.8 at xxxx Study 4.75 % 20±1°C (n=3) Report: Batch No.: Determination 100613 pH 1% w/v aqueous of the pH (BIOPREN 50 dilution = 7.2 at Values of LML 20±1°C (n=3) BIOPREN 50 mosquito LML mosquito larvicide larvicide concentrate) concentrate (after storage), Study No.: 484-122-4160 4.95% 1% w/v xxxx Study Batch No.: aqueous Report: FN/319 dilution: Determination pH = 6.6 at of the pH 20±1°C (n=3) Values of Biopren 50 LFL fly larvicide concentrate, Study No.: 484-122-0602 eCA remark: The pH reported is from a 1 %w/v solution of product Biopren 50 LFL fly larvicide concentrate. No significant differences are expected based on the composition of the products hence the reported pH value of the undiluted product after storage is considered acceptable. 20 Relative density / bulk OECD 4.95% D4 = 1.0265 xxxx Study density Guideline Batch No.: ± 0.00049 Report: 109, EPA FN/319 (with 95 % Determination OPPTS confidence of the Relative 830.7300, interval) Density of EU Method Biopren 50 LFL A.3 (Relative fly larvicide Density), concentrate, CIPAC Study No.: Handbook 484-109-0603 Volume F MT 3, Pycnometer method Storage stability test – CIPAC MT 4.95% Stored for 2 xxxx Study accelerated storage 46.3, OPPTS Batch No.: weeks at 54 °C Report: 830.6303, FN/319 in a plastic box Determination OPPTS (COEX PE/PA) of the 830.6302, Accelerated OPPTS Appearance Storage 830.6304 T=0: Viscous Stability of turquoise liquid BIOPREN 50

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Purity of Guideline the test Property Results Reference and Method substance (% (w/w) with slightly LFL fly sweetish smell larvicide T=2w: Viscous concentrate, turquoise liquid Study No.: with slightly 484-160-0533 sweetish smell

Active substance content: T=0: 4.74% w/w T=2w: 4.64% Decrease: 2.1% eCA remark : The container material is described as ‘plastic’ in the study report. The applicant specified the material as co-extruded PA/PE. As the product is aqueous, extrapolation to other plastics is acceptable. The active substance content was measured using a validated analytical method (see section ‘Methods for detection’).

An additional study in which the product BIOPREN 50 LML mosquito larvicide concentrate was tested was provided but the results were not summarized in the PAR as the applicant does not consider the study to be robust enough. In this study also the technical characteristics of the product have been tested. The applicant suggests to include as a condition of storage that the product should be stored at ambient temperature. The eCA finds this acceptable provided that the long term storage at ambient temperature supports the shelf-life of the product. Storage stability test – Validated 4.95% Stored for 4 xxxx long term storage at HPLC-UV Batch No.: years at Determination ambient temperature method FN/319 ambient of the Long- temperature in term Storage a opaque white Stability of HDPE plastic BIOPREN 50 bottle. LFL fly larvicide Appearance concentrate T=0: Viscous Study No.: turquoise liquid 484-170-0534 with slightly sweetish smell T=6m: Viscous turquoise liquid with slightly sweetish smell T=12m: Viscous turquoise liquid with slightly sweetish smell

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Purity of Guideline the test Property Results Reference and Method substance (% (w/w) T=18m: Viscous turquoise liquid with slightly sweetish smell T=24m: Viscous turquoise liquid with slightly sweetish smell T=30m: Viscous turquoise liquid with slightly sweetish smell T=36m: Viscous turquoise liquid with slightly sweetish smell T=48m: Viscous turquoise liquid with slightly sweetish smell

Active substance content: T=0: 4.66% w/w T=6m: 4.79% Increase: 2.8% T=12m: 4.60% Decrease: 1.3 % T=18m: 4.71% Increase: 1.1 % T=24m: 4.89% w/w Increase: 4.9% T=36m: 4.79% w/w Increase: 2.8% T=48m: 4.71% w/w Increase: 1.1%

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Purity of Guideline the test Property Results Reference and Method substance (% (w/w) During the 4 year storage period no significant changes were observed in the content of the active ingredient, appearance and the packaging of the test items. CIPAC MT 4.89% Before storage: xxxx Study 75.3, Batch No.: Report: Determinatio 136182 pH = 7.0 at Determination n of pH 20±1°C (n=3) of the pH Values OECD Values of No. 122, Biopren 50 Determinatio LML mosquito n of pH, larvicide Acidity and concentrate, Alkalinity, Study No.: adopted July 484-122-4159 26, 2013 4.75% after 3 years 8 xxxx Study *Batch No.: months Report: 100613 storage: Determination of the pH pH = 6.8 at Values of 20±1°C (n=3) Biopren 50 LML mosquito larvicide concentrate (after storage) Study No.: 484-122-4160 CIPAC MT 4.75% Before storage: xxxx 148 *Batch No.: The residue Determination Pourability of 100613 was found to be of the suspension R = 3.5 % and Pourability of concentrates the rinsed Biopren 50 residue R'= 0.3 LML mosquito %. larvicide concentrate, xxxx Study No.: 484-178-0546 4.75% After 2 years 7 xxxx *Batch No.: months Determination 100613 storage: of the

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Purity of Guideline the test Property Results Reference and Method substance (% (w/w) The residue Pourability of was found to be Biopren 50 R = 5.97 % LML mosquito and the rinsed larvicide residue R'= concentrate 0.37 %. (after storage) Study No.: 484-148-3436 eCA note: Data on pourability prior to and after storage has been provided for the product BIOPREN 50 LML mosquito larvicide concentrate. The amount of residue measured after storage is higher than the maximum requirement of 5%. The label shall contain the following statement: Rinse the empty container three times and add rinses to the spray tank. CIPAC MT 5.04% Before storage: xxxx 160 Batch No.: Spontaneity Determination Spontaneity 128436 of dispersion of Spontaneity of dispersion expressed as of Dispersion of mean value of of BIOPREN 50 suspension 3 replicates: LML mosquito concentrates 91% larvicide (5% dilution) concentrate Study No.: 484-168-3429 4.75% After 3 years 5 xxxx *Batch No.: months Determination 100613 storage: of Spontaneity Spontaneity of Dispersion of dispersion of BIOPREN 50 expressed as LML mosquito mean value of larvicide 3 replicates: concentrate 94.5% Study No.: (5% dilution) 484-168-3430 eCA note: Spontaneity of dispersion was shown for product BIOPREN 50 LML mosquito larvicide concentrate prior to and after storage on two different batches. This is considered acceptable as for both batches a high amount of the active ingredient was determined in dispersion. The analytical method used to determine the active substance is sufficiently validated according to SANCO 3030/99 rev. 4. CIPAC MT 5.04% Before storage: xxxx 161 Batch No.: Suspensibility Determination Suspensibilit 128436 (mean of 3 of y of aqueous determinations) Suspensibility suspension 99.6% at 4 g/L of BIOPREN 50 concentrates and 99.7% at LML mosquito 100 g/L larvicide concentrate

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Purity of Guideline the test Property Results Reference and Method substance (% (w/w) Study No.: 484-169-3431 4.75% After 3 years 5 xxxx *Batch No.: months Determination 100613 storage: of Suspensibility Suspensibility (mean of 3 of BIOPREN 50 determinations) LML mosquito 100.8% at 4 larvicide g/L and concentrate 101.2% at 100 (after storage) g/L Study No.: 484-169-3432 eCA note: Suspensibility has been sufficiently proven on the product BIOPREN 50 LML mosquito larvicide concentrate on two different batches. The analytical method used to determine the active substance is sufficiently validated according to SANCO 3030/99 rev. 4. CIPAC MT 4.75% Before storage: xxxx Wet 185 Wet *Batch No.: Wet sieve test: Sieve Test of Sieve Test 100613 0.00% Biopren 50 of retention on LML mosquito 75 µm mesh larvicide size sieve concentrate Study no: 484-164-0545 After 3 years 4 xxxx Wet months Sieve Test of storage: Biopren 50 Wet sieve test: LML mosquito 0.11% larvicide of retention on concentrate 75 µm mesh (after storage) size sieve Study no: 484-164-3435 eCA note: The wet sieve test has been performed on the product Biopren 50 LML mosquito larvicide concentrate which has an identical composition with the product BIOPREN 50 LFL fly larvicide concentrate (except colour). The results are considered acceptable. CIPAC MT 47 5.04% Before storage: xxxx Persistent Batch No.: Persistent Determination foaming 128436 foam: the of Persistent volume of foam Foaming of obtained at ca. BIOPREN 50 8% dilution LML mosquito was: larvicide 0 mL after 10 s concentrate 0 mL after 1 Study No.: min 484-158-3427

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Purity of Guideline the test Property Results Reference and Method substance (% (w/w) 0 mL after 3 min 0 mL after 12 min 4.75% After 3 years 4 xxxx *Batch No.: months Determination 100613 storage: of Persistent Persistent Foaming of foam: the BIOPREN 50 volume of foam LML mosquito obtained at ca. larvicide 8% dilution concentrate was: (after storage) 78 mL after 10 Study No.: s 484-158-3428 57 mL after 1 min 53 mL after 3 min 34 mL after 12 min eCA note: Persitent foaming was determined on BIOPREN 50 LML mosquito larvicide concentrate at an 8% dilution. No foam was observed at this concentration. As no different results are expected at lower or higher concentrations corresponding to the lowest and highest in use concentration the results are considered acceptable. eCA remark: The container material is described as ‘plastic box’ in the study report. According to the information provided by the applicant the material is opaque white HDPE. As the product is aqueous, extrapolation to other plastics is acceptable. No information on release rate was provided as the appropriate method for determining the release rate of the active substance is under development. The applicant proposes to provide the data at the renewal of the product authorisation.

The active substance content was measured using the validated analytical method (see section ‘Methods for detection’).

The physical, chemical and technical characteristics of the ZW formulation BIOPREN 50 LML mosquito larvicide concentrate have been used to support the shelf life of the product BIOPREN 50 LML fly larvicide concentrate. As the two products have an almost identical composition this is considered acceptable at this stage. A shelf life of 30 months in CO/EX and HDPE containers is supported for the product BIOPREN 50 LML fly larvicide concentrate. Storage stability test – The label will low temperature include the stability test for liquids statement: “Store product at temperatures

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Purity of Guideline the test Property Results Reference and Method substance (% (w/w) between 5 and 40 °C”. Effects on content of the During the active substance and stability study technical characteristics the effects of of the biocidal product - light, humidity light and higher temperature were not investigated as the packaging of the water- based product is closed, opaque white, water proof and ensures the protection from direct sunlight and other impacts of the environment. The label also contains the proper and detailed instruction for safe storage of the product. Effects on content of the See "Effects on active substance and content of the technical characteristics active of the biocidal product – substance and temperature and technical humidity characteristics of the biocidal product – light " results. Additionally, the conditions of storage limit the storage of the product in the temperature range of 5°C

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Purity of Guideline the test Property Results Reference and Method substance (% (w/w) of the biocidal product - that the opaque reactivity towards white HDPE container material packaging material has not reacted with the product. No changes were observed in the appearance of the product and packaging of the test item during the study. Wettability Not applicable, due to the product being a water-based liquid. Suspensibility, Please see data spontaneity and at long term dispersion stability storage at ambient temperature. Wet sieve analysis and CIPAC MT 4.95% The percentage xxxx Wet dry sieve test 185 Wet Batch No.: retention on Sieve Test of Sieve Test FN/319 the 75 µm Biopren 50 LFL mesh size sieve fly larvicide was: 0.00 %. concentrate, Additional data xxxx 2015 included at Sponsor: Storage Babolna Bio stability test – Ltd., Study long term No.: 484-164- storage at 0606 ambient temperature. Emulsifiability, re- Not applicable, emulsifiability and due to the emulsion stability formulation type (ZW) Disintegration time Not applicable, due to the formulation type (ZW) Particle size distribution, Justification According to content of dust/fines, for the non- FAO/WHO attrition, friability submission guide 7.33 Note of data 11, particle size

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Purity of Guideline the test Property Results Reference and Method substance (% (w/w) distribution “…should be evaluated by 4.5.31 Wet sieve test and 4.5.43 Suspensibility or 4.5.44 Dispersion stability.” Wet sieve test study and Suspensibility test study have been performed on BIOPREN 50 LML. Please see data at Storage stability test – long term storage at ambient temperature. eCA remark: Particle size distribution has not been directly determined but a wet sieve test and suspensibility test were determined on the product BIOPREN 50 LML. The results provided are considered satisfactory. Persistent foaming Please see data above, at point Storage stability test – long term storage at ambient temperature. Flowability/Pourability/Du CIPAC MT 4.95% The residue xxxx stability 148 Batch No.: was found to be Determination Pourability of FN/319 R = 3.2 % and of the suspension the rinsed Pourability of concentrates residue R'= 0.3 Biopren 50 LFL %. fly larvicide concentrate, Additional xxxx 2015, information on Study No.: pourability is 484-178-0607 included in section Storage stability test – long term storage at

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Purity of Guideline the test Property Results Reference and Method substance (% (w/w) ambient temperature. Burning rate — smoke Not applicable, generators the product is a water-based liquid, not a smoke generator. Burning completeness — Not applicable, smoke generators the product is a water-based liquid, not a smoke generator. Composition of smoke — Not applicable. smoke generators The product is a water-based liquid, not a smoke generator. Spraying pattern — Not applicable. aerosols The product is a water-based liquid, not an aerosol. Physical compatibility Not applicable as the product is not intented to be used as tank mix. Chemical compatibility Not applicable as the product is not intented to be used as tank mix. Degree of dissolution and Not applicable dilution stability for the type of formulation assigned to this biocidal product. Surface tension OECD 4.95% The surface xxxx Study Guideline Batch No.: tension of Report: No. 115, FN/319 Biopren 50 LFL Determination European fly larvicide of the Surface Council concentrate Tension of Regulation aqueous Biopren 50 LFL (EC) No. solution (250.2 fly larvicide 440/2008, mg / 250 mL) concentrate, Annex Part was found to be Study No.:

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Purity of Guideline the test Property Results Reference and Method substance (% (w/w) A, Methods 35.2 mN/m at 484-115-0605, for the 20 °C using the May 13, 2015 determinatio ring method. n of physico- The surface chemical tension of properties, Biopren 50 LFL A.5. fly larvicide “Surface concentrate Tension”, EC aqueous Publication solution (4.8 No. O.J. L mL/100 mL) 142/50, May was found to be 2008 32.7 mN/m at 20 °C using the ring method. As these values are lower than 60 mN/m, Biopren 50 LFL fly larvicide concentrate is classified as surface active. Viscosity OECD Test 4.95% Viscosity at xxxx Study Guideline Batch No.: 20°C Report: 114, DIN FN/319 (dynamic): Determination 53015 106.7 mPa*s. of the Viscometry Viscosity at Viscosity of (falling ball 40°C Biopren 50 LFL viscometer) (dynamic): fly larvicide 131.1 mPa*s concentrate, Study No.: 484-114-0604, June 15, 2015

* GLP Long term stabilty study of BIOPREN 50 LML mosquito larvicide concentrate (batch No.: 100613) proves the stability of the product.

Conclusion on the physical, chemical and technical properties of the product The product BIOPREN 50 LFL fly larvicide concentrate is a partly microencapsulated liquid formulation containing 50 g/kg (5.0 %w/w) S-methoprene, intended for dilution in water prior to spray application. The overall a.s. content is 5.0% w/w as pure a.s. (minimum purity: 95% w/w): 1.25% w/w as free S-methoprene and 3.75% w/w as micro-encapsulated S-methoprene. BIOPREN® 50 LFL FLY LARVICIDE CONCENTRATE is a turquoise, viscous aqueous liquid formulation, with a characteristic slightly-sweetish odour. 20 The pH of the product at 20°C is neutral. The relative density D 4 was determined to be 1.0265.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Storage stability tests were conducted on the product in its original packaging (HDPE bottle). No significant change in the packaging was observed at the end of the storage (neither accelerated nor long term).

• Accelerated storage stability test: the product in its original packaging (HDPE bottle) was kept at 54 °C for 2 weeks. No significant chemical change occurred (a.s. content variation: -2.1%); no significant change in the product appearance was observed, either. No data on acidity/alkalinity/pH range; pourability; spontaneity of dispersion; suspensibility; wet sieve test submitted, therefore the label shall contain the statement limiting the storage of the product at temperatures between 5 and 40 °C. • Long-term storage stability study: After 4 years at 20±1 °C, the a.s. content increased by 1.1% w/w. No change in the product appearance was observed. Read across data from product BIOPREN 50 LFL mosquito larvicide concentrate was submitted showing that the product remained stable upon storage (6.8 and 7.0 at 20±1 °C, for the neat formulation). Additionally, read-across results were presented for persistent foam (CIPAC MT 47), spontaneity of dispersion (CIPAC MT 160), suspensibility (CIPAC MT 184), pourability (CIPAC 148.1), and wet sieve test (CIPAC MT 185). The most relevant technical characteristics of the product proved to be still satisfactory after storage for at least 30 months, thus ensuring that BIOPREN® 50 LFL FLY LARVICIDE CONCENTRATE remains suitable for use after transportation and storage. As for pourability, results after storage were slightly out of the acceptable limits. To ensure that no excessive residues remain in the container the label contains: “Rinse the empty container three times and add rinses to the spray tank”. Taking into account the overall storage stability data, the results support a shelf-life of 30 months. Low temperature stability test was not conducted. Therefore, label contains: “Store product at temperatures between 5 and 40°C”. Testing on the effect of light is not necessary for BIOPREN® 50 LFL FLY LARVICIDE CONCENTRATE. Though the a.s. is sensitive to light, opaque packaging ensures protection from direct sunlight. So, exposure to sunlight is negligible when the product is correctly stored in its original packaging (HDPE or COEX PA/PE) according to label instructions. As BIOPREN® 50 LFL FLY LARVICIDE CONCENTRATE is a CS formulation, the following technical characteristics were addressed before storage to ensure the product is easy to handle and offers good miscibility with water: - persistent foam; - spontaneity of dispersion; - suspensibility; - pourability. The wet sieve test was also carried out, to ensure the sprayability of the diluted product. Test results showed the satisfactory performance of BIOPREN® 50 LFL FLY LARVICIDE CONCENTRATE, in particular that the product is homogeneous on application, with no residue remaining which could cause blockages of the spraying equipment. BIOPREN® 50 LFL FLY LARVICIDE CONCENTRATE is not intended to be used in combination with other products. The surface tension values (35.2 mN/m, 250.2 mg / 250 mL and 32.7 mN/m, 4.8 mL / 100 mL) determined for the product indicate that BIOPREN® 50 LFL Fly larvicide concentrate is surface active. The dynamic viscosity of the Newtonian fluid was determined to be 106.7 mPa s and 131.1 mPa s at 20°C and 40°C, respectively.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18 eCA remark: The physical, chemical and technical characteristics of the product BIOPREN® 50 LFL fly larvicide concentrate have been sufficiently addressed also taking into account the tests performed with the almost identical product BIOPREN® 50 LFL mosquito larvicide concentrate. The shelf life of 30 months in HDPE and COEX PE/PA is supported.

2.2.3 Physical hazards and respective characteristics

Guideline Purity of the test Property and substance (% Results Reference Method (w/w) Explosives Justification -- Test not -- for the non- considered to submission be conducted of data since none of the components present in the formulated biocidal product contain groups mentioned in Table A6.1 in Appendix 6 of the UN-MTC which may indicate explosive properties. Flammable gases -- -- The product -- does not need to be considered for this hazard class, as the biocidal product is not a gas. Flammable aerosols -- -- The product -- does not need to be considered for this hazard class, as the product is not an aerosol. Oxidising gases -- -- The product -- does not need to be considered for this hazard class as it is not a gas. Gases under -- -- Not relevant, -- pressure because the

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Guideline Purity of the test Property and substance (% Results Reference Method (w/w) product is not a gas under pressure. Flammable liquids EU A.9 4.95% The product xxxx (closed cup Batch No.: FN-319 has no flash Determination ASTM point up to 100 of Flash Point of D7094) °C, and above Biopren 50 LFL this fly larvicide temperature concentrate the Study No: 484- determination 150-0601 of this property is impossible because of the water vapour saturated headspace of the flash point measuring equipment. The product has more than 60% water content, and contains no flammable ingredients, according to their SDSs. According to theses facts, the product is not considered to be a flammable liquid. eCA remark: The product was tested using the closed cup method in order to determine the flash-point. No flash point was observed following the screening tests. It can be concluded that the product is not to be classified as (highly) flammable liquid according to the Reg. EU 1272/2008. Flammable solids -- -- The product -- does not need to be considered for this hazard class, because the product is a liquid. Self-reactive Justification -- None of the -- substances and for the non- components of mixtures submission the product of data contain any of

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Guideline Purity of the test Property and substance (% Results Reference Method (w/w) the listed example moieties in Guidance on the Application of the CLP Criteria 2.8 Self-reactive substances and mixtures or in Appendix 6, Section 5.1 of the UN-MTC, therefore the product is not considered to be self-reactive mixture. Pyrophoric liquids -- -- The product -- does not need to be considered for this hazard class, the product is not a pyrophoric liquid and its ingredients have no pyrophoric property. Pyrophoric solids -- -- The product -- does not need to be considered for this hazard class, the product is a liquid. Self-heating Justification -- Product is a -- substances and for the non- liquid and mixtures submission according to of data the Guidance on the Application of the CLP Criteria 2.11.7.2. Examples of substances and mixtures not fulfilling the

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Guideline Purity of the test Property and substance (% Results Reference Method (w/w) classification criteria: In general, liquids show no self-heating behaviour unless adsorbed on a large surface, therefore this test is not to be conducted. Substances and Justification -- The product -- mixtures which in for the non- does not need contact with water submission to be emit flammable of data considered for gases this hazard class, because the product contains water. Oxidising liquids Justification -- The -- for the non- components submission contain oxygen, of data but this element is chemically bonded only to carbon or hydrogen atoms. Since none of the ingredients are officially classified or self-classified as oxidizing agent, the product is not expected to possess oxidising properties. Oxidising solids -- -- The product -- does not need to be considered for this hazard class, the product is a liquid.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Guideline Purity of the test Property and substance (% Results Reference Method (w/w) Organic peroxides Justification -- No organic -- for the non- peroxides are submission present. The of data product should not be considered for classification in this hazard class. Corrosive to metals Justification -- The product -- for the non- should not be submission considered for of data classification in this class because it fullfils the following criteria mentioned in 2.5 Corrosive to metals, TAB- APCP: halogen- free, no strong base/acid, no complexing agents, pH of the neat formulation is neutral. Auto-ignition EU A.9 4.95% No flash point -- temperatures of (closed cup) Batch No.: FN-319 observed up to products (liquids and 100°C. Due to gases) the high water content of the test item the flash point determination is not applicable above 100°C using closed cup method, therefore the product will not have a flash point up to 200°C. In this case an Auto- ignition study is not required.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Guideline Purity of the test Property and substance (% Results Reference Method (w/w) EU method 5.04% No self-ignition xxxx A.16 Batch No.: 128436 was observed Determination up to 400°C of the Auto- Ignition Temperature of BIOPREN 50 LML mosquito larvicide concentrate Study No.: 484-153-3433 eCA remark: During the preliminary test the test item BIOPREN® 50 LML mosquito larvicide concentrate did not ignite hence no auto-ignition temperature could be set. The results of this testing are considered acceptable also for the product BIOPREN® 50 LFL fly larvicide concentrate. Relative self-ignition Not applicable temperature for as the biocidal solids product is a liquid . Dust explosion Not applicable hazard as the biocidal product is a liquid.

Conclusion on the physical hazards and respective characteristics of the product The product was sufficiently tested in order to conclude that it should not be classified based on the physical hazards and respectice characteristics. Since neither the a.s. nor the co-formulants are officially classified or self-classified as explosive or oxidising, it can be anticipated that BIOPREN® 50 LFL FLY LARVICIDE CONCENTRATE has neither explosive nor oxidising properties. Neither the a.s. nor the co-formulants are officially classified or self-classified as flammable, so BIOPREN® 50 LFL FLY LARVICIDE CONCENTRATE is not expected to be a flammable liquid; besides, the product is an aqueous formulation, it has no flash point up to 100°C, contains no flammable ingredients, according to their SDSs. According to this facts, the product is not considered to be a flammable liquid. No self-ignition was observed up to 400°C. There are no chemical groups associated with self-reactive properties, either. Experience in production and handling shows that BIOPREN® 50 LFL FLY LARVICIDE CONCENTRATE does not ignite spontaneously on coming into contact with air at normal temperatures. Experience in handling and use shows that the product does not emit flammable gases after contact with water, either. BIOPREN® 50 LFL FLY LARVICIDE CONCENTRATE should not be considered for classification in the hazard classes of organic peroxide and corrosive to metals, based on considerations on the a.s. and the co-formulants. On the basis of the available data/information, BIOPREN® 50 LFL FLY LARVICIDE CONCENTRATE does not pose any physical hazards. Therefore, the need for a risk characterisation for physical hazards is not envisaged.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18 eCA remark: Considering the provided test results and waivers the product BIOPREN® 50 LFL fly larvicide concentrate is not to be classified based on its physical hazards and characteristics.

2.2.4 Methods for detection and identification

A method by RP-HPLC with UV detection at 265 nm on a Luna 3 μ C18 (2) 100A column for the determination of S-methoprene in BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE has been developed and validated according to SANCO/3030/99 rev. 4. The test item is dissolved in ethanol, then sonicated three times for 10 minutes. The sample is left standing for 1 hour. Then, after repeated sonication (10 minutes), the sample is centrifuged at 20°C, at 4000 rpm (10 min) and then diluted twenty-fold with acetonitrile before analysis.

The method is: - sufficiently specific, as showed by example chromatograms from the analysis of S- methoprene standard, placebo and product sample; - linear (R 2 > 0.999) over the range 4.0–6.0% w/w, corresponding to ca. 80-120% of the nominal a.s. content; - accurate, with mean recovery rates in the acceptable range 97-103% at spiking levels of 4.25 and 5.75% w/w (corresponding to ca. 85 and 115% of the nominal a.s. content); - precise, the %RSD= 1.3% and 1.5% (n=5) is below the limit of 2.1% given by the modified Horwitz equation.

Analytical methods for the analysis of the product as such including the active substance, impurities and residues Analyte Analyti Fortificatio Linearit Specificity Recovery Limit of Referenc (type of cal n range / y rate (%) quantifica e analyte metho Number of tion e.g. d measurem Ran Me RS (LOQ) or active ents ge an D other substan limits ce) S- reverse 2 levels Linear The level lev lev Not Validation methopr phase (level 1: across a specificity 1: el el required of the ene HPLC 4.25 %; range of of the 97- 1: 1: Analytical method level 2 5.75 8 - 12 method for 101 99 1.4 Method

with UV %) μg/mL S- % (HPLC) for detectio S- Methoprene the n = 5 n (265 methopr in BIOPREN determina

nm) on ene 50 LFL fly tion of S- lev a Luna y = larvicide level lev Methopre el 3μ C18 103331x concentrate 2: el ne in 2: (2) - 8172 was 97- 2: BIOPREN5 100A examined 101 99 1.5 0 LFL fly R2 = 1 column by the % larvicide analysis of concentrat control e, Study matrices No.: 484- Placebo 100-0535 Biopren 50 Date of LFL fly Final larvicide Report: concentrate April 29, . The

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

chromatogr 2015, ams of the xxxx control solutions showed no interfering substances at the retention time (approx 5.4 min) of S- Methoprene

Analytical methods for monitoring

Analyte Analytic Fortification Lineari Specifici Recovery rate Limit of Referen (type of al range / ty ty (%) quantificati ce analyte method Number of on (LOQ) e.g. measureme Rang Mea RS or other active nts e n D limits substanc e)

The analytical methods for monitoring have been addressed in the EU assessment report. No additional data are required.

Analytical methods for soil

No analytical methods for soil are necessary for the product based on the following considerations .

According to the ECHA guidance Volume I, Part A, 2.8.2.1 Point 5.2.1 Soil: “If the active substance degrades very quickly, i.e. DegT 50 and DegT 90 values of the active substance and the relevant metabolites are lower than two and three days ; respectively, analytical methods for residues in soil are not required except in the case of continuous exposure.” eCA remark: The active substance is applied encapsulated and hence it is possible that it does not degrade very quickly. In this case the DegT 90 can be higher than 3 days. Evidence should be provided that the DegT 90 is lower than 3 days.

For S-methoprene and its metabolites, none of the criteria above are met . Therefore, no validated analytical method is needed for S-methoprene monitoring.

According to the guidance, in general an environmental risk assessment for the relevant compartments needs to be performed for major metabolites. If there is any reason for concern, a risk assessment also needs to be performed for minor metabolites which are ecotoxicologically relevant. For S-methoprene, no major metabolites have been identified in the soil degradation study.

Minor metabolites lack any environmental relevance as S-methoprene’s mode of action is an insect growth hormone regulator. After degradation, the degradation product will lose

NL Biopren 50 LFL FLY Larvicide concentrate PT 18 that mode of action. According to the degradation pathway described in the soil biodegradation study report, the identified degradation products are intermediates and they are further degrading to “several minor and transient fractions” and as a terminal degradation, significant CO 2 production was measured. Therefore, the degradation products are not relevant for environmental risk assessment. Monitoring is not relevant in soil, consequently an analytical method in soil is not necessary.

Analytical methods for air

Based on the vapour pressure of S-methoprene there is not expected that the active ingredient appears in the air. The fugasity modelling confirms the same, as it shows 0% presence in the air. Nevertheless, for S-methoprene-based products to be sprayed, only coarse sprays with big droplets (MMAD >50 μm) are recommended, so the inhalable amount of product is expected to be negligible.

Analytical methods for animal and human body fluids and tissues

The practical using concentration of S-methoprene is extremely low compared to NOAEL and LOAEL values in animal tests. So the exposure and the result in residue in animals are negligible. Therefore no special analytical method was developed for monitoring purpose.

Analytical methods for monitoring of active substances and residues in food and feeding stuff Analyte Analytic Fortification Lineari Specifici Recovery rate Limit of Referen (type of al range / ty ty (%) quantificati ce analyte method Number of on (LOQ) e.g. measureme Rang Mea RS or other active nts e n D limits substanc e)

eCA remark: The analytical method for monitoring of active substances and residues in food and feeding stuff has not been performed according to SANCO/825/00 rev 8.1. Based on the specific use of the product analytical methods for determination of S-Methoprene in food and feeding stuffs are not required.

Conclusion on the methods for detection and identification of the product Validation of the analytical method for the determination of S-methoprene in BIOPREN 50 LFL fly larvicide concentrate was performed. In the course of this study has been validated a reverse phase HPLC method with UV detection on a Luna 3 μ C18 (2) 100A column. The procedure was found to be suitable for the analysis.

Analytical methods for air

Because the active is non volatile and based on the use pattern, a method of analysis in air is not required.

Analytical methods for animal and human body fluids and tissues

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

S-methoprene is not classified as being toxic or highly toxic. It is therefore proposed in the Assessment Report of the active substance that analytical methods in animal and human body fluids and tissues are not required. Nevertheless validated method is available for the determination of S-Methoprene in Edible Meat Matrices. The method is reserve phase HPLC method with UV detection on a Luna 3μ C18 (2) 100A column.

2.2.5 Efficacy against target organisms

2.2.5.1 Function and field of use BIOPREN 50 LFL fly larvicide concentrate is an Insect Growth Regulator within the group of insecticides (PT18) which is primarily effective against Musca domestica. This larvicide is to be used both indoors and outdoors by (trained) professionals only. Typical treatment areas are different types of manures in stables (indoor) of different species (pig andcow) in order to considerably decrease the number of larvae and to stop their development into imagoes. Outdoor use only concerns the treatment of manure/slurry kept in a concrete basin (leak- proof insulated).

2.2.5.2 Organisms to be controlled and products, organisms or objects to be protected Target organisms: Musca Domestica – House fly - larvae Stomoxys calcitrans – Stablefly - larvae Eristalis tenax – Dronefly - larvae

2.2.5.3 Effects on target organisms, including unacceptable suffering BIOPREN 50 LFL is an S-methoprene based insect growth regulator, a juvenile hormone analogue. The use of the product makes it possible to eliminate the less visible part of a fly population which represent more than 80 % of the full population The product prevents development of the larvae into the adult reproductive stages.

2.2.5.4 Mode of action, including time delay

Juvenile hormone analogue (JHA) insecticides impair the endocrine system of insects by blocking larval–pupal transformation and pupal-adult metamorphosis. S-methoprene acts as a JH agonist, it mimics the action of JH III. Juvenile hormone is normally produced by larvae and modulates the action of ecdysone burst which occurs at larval molts leading to maintaining larval development. During the early phases of metamorphosis, JH level decreases and thus, the effect of ecdysone can be expressed leading to metamorphic changes. If external JHA is applied during early metamorphosis, it binds to JH-interacting proteins forming a complex which alters the expression of early ecdysone-regulated metamorphic genes required for normal developmental changes. This results in developmental disruption: failure in egg hatching (probably due to the impairment of early embryonic development), increased mortality during pupal development, and - in some species - sterility in the emerged adults. The presence of external JHAs during the last instar phase can also result in the formation of non-viable larval–pupal or pupal–adult intermediates. Since JHA-type insecticides affect insect development only at specific, susceptible developmental stages, it takes some time before their effects are clearly visible at the level of the entire insect population.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Treated larvae continue to develop to the pupal stage after which they fail to emerge. Expected time delay from the beginning of application to manifestation of the biocidal effect is 2-4 weeks (depending on severity of the infestation and life cycle of the target insect) based on the results of efficacy studies and life cycle of Diptera species.

Methoprene is an invertebrate metabolic inhibitor which does not seem to cause direct toxic effects in mammals. (IPCS INCHEM / WHO, No 47, Methoprene). It modulates ecdysteroid signalling during insect development and metamorphosis. S-methoprene is an analogue to a unique insect-growth regulating hormone, which does not resemble any known mammalian hormones. Applied at very low rates, while insect populations are still in the egg or larval stage of their life cycle, methoprene prevents development to the adult reproductive stages so that insects die in arrested immaturity.

S- Methoprene is not toxic when applied to adult stages of the target insect. Because the chemical interferes with the normal life cycle of insects and is not directly toxic to the pest, it is considered to be a biochemical pesticide (EXTOXNET 2001).

2.2.5.5 Efficacy data

Experimental data on the efficacy of the biocidal product against target organism(s) Function Field Test Test Test Test system / Test Referenc of use substance organism metho concentrations results: e envisa (s) d applied / effects ged exposure time Larvicide indoor BIOPREN Musca Field 12 buildings Biopren 50 FIELD 50 LFL - 5% Domestica test. were monitored: LFL: TESTING (w/w) (larvae) French 1 - 4: treatment OF AN methoprene registra with Mean of INSECTIC tion experimental flies IDE standar product (2 before SPECIALI Device SC d cowshed and 2 treatment: TY method sheepfold) INTENDE 15 (150 g/l 268 (416, diflubenzur ology 4 -8: treatment D TO 392, 87 on) was C.E.B. with reference CONTROL used as a N°107 product (3 and 179). FLIES IN reference cowshed and 1 BREEDIN product sheepfold) Reduction: G 9 -12: untreated 2.4, 48.5, PREMISE control (1 81.4, 90.5 S cowshed, 2 and 91.9% Report n° sheepfold and 1 (average) 1824/091 pigsty) after 1, 4 2,,4, 8 and xxxx Dosage of tested 12 weeks product: dilution respectively in water at a . rate of 0.04%, then 50 ml/m2 Reference of the water product: mixture = 200 ml of product in Mean of 5 L of water flies applied on 100 before m2. treatment:

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

133 (69, Dosage of the 143, 205 reference and 116) product: 5 kg a.i./ha (3.33 g Reduction: product /m2 ) 0,1, 50,5, with a water 78,4, 89,3 rate of 5000 and 91,3% l/ha. (average) after 1, Treatment: The 2,,4, 8 and product was 12 weeks applied as a respectively spray in the . empty buildings (no animals) on Control manure pits or treatment: grids where the larvae occur. Mean of Only one flies application was done. before treatment: Test period: 76 (108, September – 80, 49 and December 2014. 67)

Reduction: 2.9, 0,9, - 3.5, -4.5 and 0.2% (average) after 1, 2,,4, 8 and 12 weeks respectively . Larvicide indoor Name: Musca Labor Larvae: 4 day EFFICACY Biopren 50 Domestica atory old L3 larvae Mortality STUDY LFL fly (larvae) were used, 3 x OF larvicide test 50 larvae per Pig slurry: BIOPREN concentrate In- dose. 98.58 (dose 50 LFL A.i.: 5.0 ww house 1), 98.55 FLY % S- trial Medium: pig (dose 2) LARVICID methoprene proto slurry, pig and 10.46 E manure, chicken (control) % CONCENT col manure and RATE IN chicken litter, Pig DIFFEREN Any larvae manure: T present in the 94.51 (dose MANURES manure were 1), 100 IN eliminated by (dose 2) BIOLGICA storing in a deep and 2.84 L fridge for 14 (control) % LABORAT days ORY Chicken 153.043.- Conditions: 23- manure: 044.-045. 25⁰C, 50-60% 47.13 (dose 153.048.- relative humidity 1), 63.77 049.-050. (dose 2) 153.053.- 054.-055.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Dose 1: 2 ml and 6.33 153.058. - Biopren 50 LFL (control) % 059.-060. + 48 ml tap xxxx water (0.47 ml Chicken of this solution litter: 17.27 per 93.27 cm 2). (dose 1), 19.06 (dose Dose 2: 2.4 ml 2) and 5.74 of Biopren 50 (control) % LFL + 47.6 ml of tap water (0.47 Inhibition ml of this solution per per Pig slurry: 93.27 cm 2). 98.41 (dose 1), 98.38 Numbers of (dose 2) pupae and and 10.46 imagoes were (control) % recorded to determine Pig (inhibition of) manure: emergency rates 94.35 (dose and mortality 1), 100 rates (dose 2) and 2.84 Test duration: (control) % 14 days. Chicken manure: 43.56 (dose 1), 61.32 (dose 2) and 6.33 (control) %

Chicken litter: 12.23 (dose 1), 14.13 (dose 2) and 5.74 (control) % Larvicide indoor BIOPREN Stomoxys Field 12 buildings Results for FIELD 50 LFL fly calcitrans , test. were monitored Stomoxys TESTING larvicide Eristalis French for each target calcitrans OF AN concentrate tenax registra species: INSECTIC - 5% (w/w) (larvae) tion Dose 1: IDE S- standar Stomoxys 1.1, 36.1, PRODUCT methoprene d calcitrans 1 - 4: 66, 91.0 INTENDE protoco treatment with and 87.1% D TO l dose 1 of reduction CONTROL Method product (average) FLIES IN C.E.B. (1 cowshed and after 1, 2, BREEDIN n°107 3 sheepfold) 3, 8 and 12 G weeks PREMISE 4 -8: treatment respectively S TRIAL with dose 2 of . AGAINST product (2 THE cowshed, 1 Dose 2: STABLE sheepfold and 1 2.7, 59, FLY pigsty) 84.8, 94.9 Stomoxys

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

and 90.6% calcitrans 9 -12: untreated reduction AND THE control (average) DRONE (2 cowshed and after 1, 2, FLY 2 pigsty) 3, 8 and 12 Eristalis weeks tenax Eristalis tenax respectively 1931- 1 - 4: treatment . FFT50LFL with dose 1 of /0515. product Control xxxx (2 cowshed, 1 treatment: sheepfold and 1 -6.8, -6.9, - pigsty) 5.3, 1.2 and 7.9% 4 -8: treatment reduction with dose 2 of (average) product (2 after 1, 2, cowshed and 2 3, 8 and 12 pigsty) weeks respectively 9 -12: untreated . control (2 cowshed, 1 Results for sheepfold and 1 Eristalis pigsty) tenax

Dosage of tested Dose 1: product: 0.8, 41.8, 64.1, 85.7 Dosage1: 2.3 and 78.7% ml in 47.7 ml of reduction water per m² (average) after 1, 2, Dosage 2 = 3.9 3, 8 and 12 ml in 46.1 ml of weeks water per m². respectively . Treatment: the product was Dose 2: applied as a 4.1, 53.2, spray in the 86.4, 96.8 empty buildings and 93.7% (no animals) on reduction manure (average) pits or grids after 1, 2, where the larvae 3, 8 and 12 occur.. weeks Only one respectively application was . done. Control Testing period: treatment: August – -10.0, - November 2015 14.8, -4.1, -1.4 and 9.7% reduction (average) after 1, 2, 3, 8 and 12

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

weeks respectively . Larvicide indoor BIOPREN Stomoxys Laborat Dose: 2 mLl in For both Laborator 50 LFL fly calcitrans , ory 50 mL water/m 2 Stomoxys y larvicide Eristalis test. (equivalent to calcitrans measure concentrate tenax 230-240 mL in and Eristalis ment of - 5% (w/w) (larvae) 5 L water for tenax the S- reduction of effectiven 100 m 2). methoprene emergence ess of an For both was 100% IGR- species: after 0, 4, 8 insecticid 25 last instar and 12 e larvae from a weeks. speciality colony breeding. intended

Emergence for the Medium: in the control of cowshed straw controls stable + manure varied from flies

89 to 97% Stomoxys Conditions: for calcitrans 24+/-1 °C, 65 Stomoxys and +/-5 % RH, light calcitrans Eristalis 1200 lux 8 and from tenax hours + 16 82 to 88% hours darkness, xxxx for Eristalis no ventilation. 2069d/04 tenax . 16R After 0, 4 and 8 and 12 weeks, the emerged adults were counted in the treated and the untreated series.

Four replicates were conducted per series. Larvicide indoor Biopren 50 Musca Laborat Larvae: 4 day BIOLGICA LFL fly Domestica ory old L3 larvae L larvicide (larvae) test were used, 3 x Mortality LABORAT concentrate In- 50 larvae per ORY A.i.:5.0 ww house dose. Chicken EFFICACY % S- trial Medium: chicken litter: 78.34 STUDY methoprene protoco manure and (dose 1), OF l chicken litter. 78.18 (dose BIOPREN Any larvae 2) and 6.22 50 LFL present in the (control) % FLY manure were LARVICID eliminated by Chicken E storing in a deep manure: CONCENT fridge for 14 78.65 RATE IN days (dose 1), DIFFEREN 87.25 (dose T Conditions: 23- 2) and 8.81 CONSIST 25⁰C, 50-60% (control) % ENCY relative humidity MANURES Inhibition xxxx 2016,

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Dose 1: 2 ml Chicken 163.089 - Biopren 50 LFL litter: 76.90 094. + 48 ml tap (dose 1), xxxx water (0.47 ml 76.73 (dose of this solution 2) and 6.22 per 93.27 cm 2). (control) %

Dose 2: 2.4 ml Chicken of Biopren 50 manure: LFL + 47.6 ml of 76.59 tap water (0.47 (dose 1), ml of this 86.02 (dose solution per 2) and 8.81 93.27 cm 2). (control) %

Numbers of pupae and imagoes were recorded to determine (inhibition of) emergency rates and mortality rates.

Test duration: 14 days.

Conclusion on the efficacy of the product Three laboratory and two field studies were provided to demonstrate the efficacy of the product to be authorized. All test were conducted with the product to be authorized.

It can be concluded that the product is sufficiently efficacious against larvae of Musca domestica, Stomoxys calcitrans and Eristalis tenax at an application rate of 2.3-2.4 g/m2 (230-240 ml of BIOPREN 50 LFL concentrate diluted to 5 litres with water to treat 10m2 surface). Maximum sprayer flow rate: 0.3-0.5L/min. Efficacy has been demonstrated up to 12 weeks after treatment in cowsheds, pigsties and sheepfolds. Expected time delay from the beginning of the application to manifestation of the biocidal effect is 2-4 weeks.

Efficacy against chicken manure and litter, horse manure and goat manure is not authorised.

For a full evaluation of the label claims, please refer to section 2.2.5.8.

2.2.5.6 Occurrence of resistance and resistance management To prevent the development of resistance it is advised to use insecticides with different mode of action (rotation) as well in the pest control program.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Based on the Arthropod Pesticide Resistance Database ( www.pesticideresistance.org ) and other scientific databases, the occurrence of resistance against the active substance in the target species was reported in the following cases: - Musca domestica : lab selected resistance from mild, field evolved resistance was reported against methoprene (in: Kristensen M. and Jespersen J.B. (2003) Larvicide Resistance in Musca domestica (Diptera: Muscidae) Populations in Denmark and Establishment of Resistant Laboratory Strains. Journal of Economic Entomology 96 (4): 1300-1306). Field evolved and lab selected resistance against other IGRs, e.g. diflubenzuron, cyromazine and pyriproxyfen was also reported. Low level of resistance against diflubenzuron, methoprene, novaluron, pyripoxyfen, and triflumuron was demonstrated in wild house fly populations (in: Cetin H., Erler F. and Yanikoglu A. (2009) Survey of insect growth regulator (IGR) resistance in house flies (Musca domestica L.) from southwestern Turkey. Journal of Vector Ecology 34 (2): 329-37). - Stomoxys calcitrans : no reported resistance against (S)-methoprene or other IGR- type active substances (neither against hormonal IGRs, nor against chitin-synthesis inhibitors). - Erystalis tenax : no reported resistance against (S)-methoprene or other IGRs.

Considering the possible biochemical mechanism of the development of IGR resistance in Musca domestica , it can not be excluded, that the same mechanism may be evolved in other Diptera species as well. However, up to now direct evidences were not reported for the presence of S-methoprene resistance in Stomoxys calcitrans and Erystalis tenax . To prevent the development of insecticide resistance in any target species, the following recommendations for resistance management should always be observed by applying the product:

The product is intended to be used mainly indoors, at more or less enclosed, limited locations. Area of use specified in the label claims as well as specific mode of action of the active substance resulting in the lack of adult, mature insects prevents spreading of affected insects and breeding with wild, naive insect populations which also helps to avoid the development of insecticide resistance at wide range. However, observing the following rules while applying the product is recommended to prevent the development of insecticide resistance: - Products should always be used in accordance with label instructions. - It is advised to use insecticides with different mode of action in rotation during the pest control program, especially if an extended period of control is required. - Where possible, chemical treatments should be recommended to be combined with application of non-chemical measures (e.g. sticky traps or fly traps with UV lamps ). - Sufficient elimination of pest insects should be attempted in infested areas. - Levels of effectiveness should be monitored, and instances of reduced effectiveness should be investigated for possible evidence of resistance in the frame of IPM programs.

2.2.5.7 Known limitations There are no known limitations on efficacy.

2.2.5.8 Evaluation of the label claims

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

For a product intended as a larvicide, laboratory tests and simulated-use tests should be provided. Simulated-use tests can be waived if a robus field trial is submitted. For this product three laboratory and two field studies were provided.

Laboratory studies:

Three laboratory studies were provided to demonstrate efficacy against Musca domestica , Stomoxys calcitrans and Eristalis tenax . The doses used are expressed as the undiluted amount of product that is applied per square meter. The relative density of the product is 1.02, for easier assessment doses in g/m 2 and ml/m 2 are considered to be equivalent. The results of these studies can be summarized as follows:

Laboratory test with Musca domestica (xxxx) Medium Dose Mortality (%) Chicken litter 2 g/m 2 78.34 Chicken litter 2.4 g/m 2 78.18 Chicken manure 2 g/m 2 78.65 Chicken manure 2.4 g/m 2 87.25

Laboratory test with Musca domestica (xxxx) Medium Dose Mortality (%) Chicken litter 2 g/m 2 17.27 Chicken litter 2.4 g/m 2 19.06 Chicken manure 2 g/m 2 47.13 Chicken manure 2.4 g/m 2 63.77 Pig slurry 2 g/m 2 98.58 Pig slurry 2.4 g/m 2 98.55 Pig manure 2 g/m 2 94.51 Pig manure 2.4 g/m 2 100

Laboratory test with Stomoxys calcitrans and Eristalis tenax (xxxx) Species Medium Dose Reduction Reduction Reduction Reduction in in in in emergence emergence emergence emergence (%) – day (%) – 4 (%) – 8 (%) – 12 0 weeks weeks weeks Stomoxys Cowshed 2 100 100 100 100 calcitrans straw + ml/m 2 manure Eristalis Cowshed 2 100 100 100 100 tenax straw + ml/m 2 manure

According to the evaluation criteria of the BPR efficacy guidance, 90% larval mortality should be achieved in laboratory studies to demonstrate efficacy. For Stomoxys calcitrans and Eristalis tenax >90% reduction in emergence (which can be considered equivalent to mortality for IGR-based products) was demonstrated in laboratory tests at an application rate equivalent of 2 g/m 2.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

For Musca domestica >90% mortality was demonstrated at an application rate equivalent to 2 g/m 2 in pig slurry and pig manure. Sufficient efficacy was not demonstrated in chicken manure and chicken litter.

Field studies:

Two field studies were provided to demonstrate efficacy against Musca domestica , Stomoxys calcitrans and Eristalis tenax . The results of these studies can be summarized as follows:

Field test with Musca domestica (xxxx ) Dose Reduction Reduction Reduction Reduction Reduction of fly of fly of fly of fly of fly population population population population population (%) – 1 (%) – 2 (%) – 4 (%) – 8 (%) – 12 week weeks weeks weeks weeks 2 2.4 48.5 81.4 90.5 91.9 g/m 2

Although the field test with Musca domestica was performed in the months September- December, outside and inside conditions were suitable for fly reproduction. Indoor temperature and humidity is always higher in livestock buildings due to the presence of large animals in huge number. Fly infestation levels were monitored prior to the trial, and it proved the presence of an established infestation at all the selected test sites. Moreover, natural fly controlling factors (e.g. insectivorous birds and spiders) are less active or even not present in the autumn.

Two of the test sites for Biopren 50 LFL were treated before with either Solfac or Pyrethrum. However, the product was applied as a spray in the empty buildings (no animals) on manure pits or grids which are the places where the larvae reside. The presence of larvae (fly infestation) prior to the trial with Biopren 50 LFL Fly implies that residual effect of previous treatments has no significant influence on outcome of this trial. Moreover, fly population reduction rate was the same at all the parallel test sites (either pre-treated or not), outlier data were not observed indicating the existence of similar test conditions at all the parallel test sites.

According to the evaluation criteria of the BPR guidance, simulated use tests (or field tests) should show a decrease in number of emerging flies. For Musca domestica a decrease in number of flies emerging was demonstrated in the field test after 2 weeks and up to 12 weeks at an application rates of 2 g/m 2.

Field test with Stomoxys calcitrans and Eristalis tenax (xxxx ) Species Dose Reduction Reduction Reduction Reduction Reduction of fly in in in in population emergence emergence emergence emergence (%) – 1 (%) – 2 (%) – 3 (%) – 8 (%) – 12 week weeks weeks weeks weeks Stomoxys 2.3 1.1 36.1 66.0 91.0 97.1 calcitrans ml/m 2

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Stomoxys 3.9 2.7 59.0 84.8 94.9 90.6 calcitrans g/m 2 Eristalis 2.3 0.8 41.8 64.1 85.7 78.7 tenax g/m 2 Eristalis 3.9 4.1 53.2 86.4 96.8 93.7 tenax g/m 2

For Stomoxys calcitrans and Eristalis tenax a decrease in number of flies emerging was demonstrated in the field test after 2 weeks and up to 12 weeks at application rates of 2.3 and 3.9 g/m 2.

Efficacy against fly larvae:

It can be concluded that the product is sufficiently efficacious against larvae of Musca domestica , Stomoxys calcitrans and Eristalis tenax at an application rate of 2.3-2.4 g/m2. Dosage to be applied to treat 100 m2 surface: 230-240 ml of BIOPREN 50 LFL concentrate diluted to 5 litres with water. Maximum sprayer flow rate: 0.3-0.5L/min. Efficacy has been demonstrated up to 12 weeks after treatment in cowsheds, pigsties and sheepfolds Expected time delay from the beginning of the application to manifestation of the biocidal effect is 2-4 weeks.

For proper penetration and even distribution of the active substance in the whole quantity of the manure, it is important to treat every fresh manure layer accumulating on the top of previously treated layers. Since fly larvae usually live in the upper 5-15 cm of the manure, burying themselves from the top of the manure where the adults lay eggs, this is the thickness of fresh manure layer which should be treated from the top. Dosage were calculated based on these data to ensure that each set of new larvae receive the same active substance dose. This approach is in accordance with instructions of the WHO guideline (Vector control – Methods for Use by Individuals and Communities, 1997, Chapter 6: Houseflies): „Larvicides are applied with a sprayer or a watering can as emulsions, suspensions or solutions. The dosage has to be sufficient to wet the upper 10– 15 cm of the substrate, i.e. 0.5–5 litres/m2.”

As sufficient efficacy of the product was not demonstrated in chicken manure and chicken litter in the laboratory tests and none of the replicates in the field tests with the product to be authorized were performed on poultry farms, the product cannot be authorized for use on poultry farms or on chicken manure stored in bassins. As no tests at all were provided with horse or goat manure, the product can also not be authorised for use in those kind of stables.

Although authorisation is granted for both indoor and outdoor use, both field tests were conducted indoors. As the product is used directly on the manure, it seems unlikely that the location of this manure (indoor or outdoor) will influence efficacy of the product in the manure. If the location would however influence the efficacy, indoor temperature and humidity is higher in livestock buildings due to the presence of large animals in huge number. This would make it worst case compared to outside conditions for fly infestations. Therefore, the eCA considers efficacy sufficiently shown for both indoor and outdoor use.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

2.2.5.9 Relevant information if the product is intended to be authorised for use with other biocidal product(s) This biocidal product is not intended to be used in combination with other biocidal products.

2.2.6 Risk assessment for human health

2.2.6.1 Assessment of effects on Human Health Biopren 50 LFL FLY Larvicide Concentrate was not a reference product included in the CAR. No studies with the product are submitted. The classification and labelling of the formulation has been prepared based on the calculation method described in Annex I of Regulation 1272/2008/EC (for details please refer to the confidential annex of the PAR).

Skin corrosion and irritation

Conclusion used in Risk Assessment – Skin corrosion and irritation Value/conclusion BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE is not skin irritant Justification for the S-methoprene is not skin irritant and does not require value/conclusion classification for dermal irritation. Based on the criteria defined in CLP Regulation (EC) No. 1272/2008, the co-formulants are present in the biocidal product at concentrations insufficient to trigger the classification of the product. Consequently, based on the criteria defined in CLP Regulation (EC) No. 1272/2008, the product is not a skin irritant or corrosive. Classification of the No classification required according to CLP. product according to CLP

Data waiving Information Skin corrosion and irritation test on the product. requirement Justification Testing on the product/mixture does not need to be conducted if there are valid data available on each of the components in the mixture sufficient to allow classification of the mixture according to the rules laid down in Regulation (EC) No 1272/2008 (CLP), and synergistic effects between any of the components are not expected.

Eye irritation

Conclusion used in Risk Assessment – Eye irritation

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Value/conclusion No risk identified for BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE in relation of Eye irritation Justification for the The active ingredient S-methoprene is not an eye irritant. The co- value/conclusion formulants are present in the biocidal product at concentrations insufficient to trigger the classification of the product. Classification of the No classification required according to CLP product according to CLP

Data waiving Information Eye irritation test on the product requirement Justification Testing on the product/mixture does not need to be conducted if there are valid data available on each of the components in the mixture sufficient to allow classification of the mixture according to the rules laid down in Regulation (EC) No 1272/2008 (CLP), and synergistic effects between any of the components are not expected.

Respiratory tract irritation

Conclusion used in the Risk Assessment – Respiratory tract irritation Value/conclusion No risk identified for BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE in relation to respiratory tract irritation Justification for The active ingredient S-methoprene is not an irritant for the the conclusion respiratory tract. The co-formulants that are present in the biocidal product do not trigger classification of the product. Classification of No classification required according to CLP the product according to CLP

Data waiving Information Respiratory tract irritation test on the product requirement Justification Testing on the product/mixture does not need to be conducted if there are valid data available on each of the components in the mixture sufficient to allow classification of the mixture according to the rules laid down in Regulation (EC) No 1272/2008 (CLP), and synergistic effects between any of the components are not expected.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Skin sensitization

Conclusion used in Risk Assessment – Skin sensitisation Value/conclusion BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE is classified with H317: May cause an allergic skin reaction Justification for the The active ingredient S-methoprene is not senzitizing. One value/conclusion component classified for H317 is present in a concentration that triggers classification, Polyethylene glycol (15)-hydroxystearate. For the exact concentration and calculation see confidential composition annex. Classification of the H317: May cause an allergic skin reaction product according to CLP

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Data waiving Information Skin sensitisation test on the product requirement Justification Testing on the product/mixture does not need to be conducted if there are valid data available on each of the components in the mixture sufficient to allow classification of the mixture according to the rules laid down in Regulation (EC) No 1272/2008 (CLP), and synergistic effects between any of the components are not expected.

Respiratory sensitization (ADS)

Testing on the product/mixture does not need to be conducted if there are valid data available on each of the components in the mixture sufficient to allow classification of the mixture according to the rules laid down in Regulation (EC) No 1272/2008 (CLP), and synergistic effects between any of the components are not expected. The active ingredient S-methoprene is not have respiratory sensitizing properties. Furthermore, the co-formulants that are present in the biocidal product do not trigger classification of the product.

Acute toxicity Acute toxicity by oral route

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Value used in the Risk Assessment – Acute oral toxicity Value Not required Justification for The active ingredient S-methoprene is not classified with acute oral the selected toxicity. The co-formulants classified with acute oral toxicity are value present in the biocidal product at concentrations insufficient to trigger the classification of the product. Classification of No classification required according to CLP the product according to CLP

Data waiving Information Acute oral toxicity test on the product requirement Justification Testing on the product/mixture does not need to be conducted if there are valid data available on each of the components in the mixture sufficient to allow classification of the mixture according to the rules laid down in Regulation (EC) No 1272/2008 (CLP), and synergistic effects between any of the components are not expected.

Acute toxicity by inhalation

Value used in the Risk Assessment – Acute inhalation toxicity Value Not required Justification for The active ingredient S-methoprene is not classified with acute the selected inhalation toxicity. The co-formulants classified with acute oral toxicity value are present in the biocidal product at concentrations insufficient to trigger the classification of the product. Classification of No classification required according to CLP the product according to CLP

Data waiving Information Acute inhalation toxicity test on the product requirement Justification Testing on the product/mixture does not need to be conducted if there are valid data available on each of the components in the mixture sufficient to allow classification of the mixture according to the rules laid down in Regulation (EC) No 1272/2008 (CLP), and synergistic effects between any of the components are not expected.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Acute toxicity by dermal route

Value used in the Risk Assessment – Acute dermal toxicity Value Not required Justification for The active ingredient S-methoprene is not classified for acute dermal the selected toxicity. The co-formulants are also not classified with acute dermal value toxicity. Classification of No classification required according to CLP the product according to CLP

Data waiving Information Acute dermal toxicity test on the product requirement Justification Testing on the product/mixture does not need to be conducted if there are valid data available on each of the components in the mixture sufficient to allow classification of the mixture according to the rules laid down in Regulation (EC) No 1272/2008 (CLP), and synergistic effects between any of the components are not expected.

Information on dermal absorption Dermal absorption can be estimated on the basis of existing information that comes from other sources. Study results from an in-vitro study using human volunteer skin with radiolabeled active ingredient S-methoprene is available that may give a good estimate.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Summary table of in vitro studies on dermal absorption Method, Species, Test Absorption data Remarks Reference Guideline, Number of substance, for each (e.g. GLP status, skin Doses compartment and major Reliability samples final absorption deviations) tested per value dose, Other relevant information about the study OECD 428, Human skin, Test Total dislogeable: xxxx GLP, one dose, formulation: 94.16 % Reliability 1 total of 10 Blank Dermal delivery: skin samples Bioprene 1.61 % Pharaoh’s Stratum corneum Ant Colony (tape strips 6-20): Eliminator + 1.21% 0.5 % w/w Strip 3-5: 0.58 % labeled S- methoprene Dermal absorption of , Test [14C] -S- preparation: methoprene is 3.44 0.125 ≈ 3.5%. w/w% S- methoprene

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Value(s) used in the Risk Assessment – Dermal absorption Substance S-methoprene Value(s)* 25% (concentrate), 50% (dilution) Justification for An agreed dermal adsorption rate of 3.50% was established in the AR the selected an in vitro study using human volunteer skin and radio-labelled S- value(s) methoprene (0.125% w/w).

The product contains 5.26% (technical) S-methoprene. For its use, the concentrate’s max. dilution is 240 ml BIOPREN 50 LFL concentrate diluted with water to 5 litres, equal to 0.26% S- methoprene. Furthermore, the product is not classified for dermal irritation. The value of 3.5% is a reasonable estimate for the dermal absorption of Biopren 50 LFL FLY Larvicide concentrate. Nevertheless, as no dermal absorption study is available with the product Biopren 50 LFL FLY Larvicide, it is proposed to use the default values of the EFSA guidance on dermal absorption (2017) in the exposure assessment calculations as an absolute worst case approach. For a CS formulation, 25% absorption is indicated for the concentrate and 70% for the dilution. As the latter value results in irrealistic and largely overestimated and unacceptable risks, it is proposed to use the 50% default for water-based products (EFSA 2017) for the dilution, considering that the product is diluted approximately 20 times (i.e. 240 ml BIOPREN 50 LFL concentrate diluted with water to 5 litres) and the dermal absorption on the CAR did not exceed 5% (i.e. 3.5% is reported in the CAR, performed with a different product). However, considering that the dermal absorption as reported in the CAR did not exceed 5%, the use of a 50% dermal absorption value is already 10 times higher and thus considered worst case. Additionally, the product is diluted approximately 20 times (i.e. 240 ml BIOPREN 50 LFL concentrate diluted with water to 5 litres). Due to this dilution, the co-formulant Polyethylene glycol (15)- hydroxystearate responsible for the sensitising properties is diluted to a concentration that would trigger EUH208 classification. EUH208 classification is considered a hazard communication to persons already sensitised to a specific substance that is indicated in the sentence of EUH208. Therefore, for risk assessment purposes, the dermal absorption of the in-use dilution is considered not affected by the classification and a value of 50% is considered applicable for the active substance exposure calculations.

Data waiving Information Dermal absorption test on the product requirement Justification Dermal absorption can be estimated on the basis of existing information and default values that comes from other sources (see detailed explanation above).

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Available toxicological data relating to non active substance(s) (i.e. substance(s) of concern)

Two substances of concern in the product are identified in accordance to the guidance (CA- Nov14-Doc.5.11). One of the SoCs is Polyethylene-glycol-15-hydroxy-stearate as it causes the classification of the product with: H317: May cause an allergic skin reaction. This classification is dedicated to band B, and therefore a qualitative exposure and risk assessment to determine whether P-statement(s) normally associated with concerned H statement is sufficient needs to be performed.

Based on this classification with H317 and taking into account the product is applied by spraying, P280: Wear protective gloves and protective clothing is prescribed. Taking this into account, risk due to this hazard is considered mitigated. Furthermore, the following general risk mitigation measures are included in the SPC: ‘Avoid contact with skin and eyes’ and ‘Wash hands thoroughly with hot water and soap after use’. No additional risk mitigation measures are considered needed.

Biopren 50 LFL FLY Larvicide concentrate contains < 1% ethylene glycol (CAS 107-21-1). For ethylene glycol, there is an European workplace exposure limit available. Therefore, according to the Substance of Concern Guidance, CA-Nov14-Doc5.11, point 5 of “Identification of Substances of Concern”, ethylene glycol is to be considered as substance of concern and a quantitative risk assessment is required.

According to the Scientific Expert Group on Occupational Exposure Limits, a 8-hour-TWA of 52 mg/m 3, a short-term exposure level STEL (15 mins) of 104 mg/m 3 and a “Skin Notation” is recommended for ethylene glycol.

However, primary exposure by inhalation is possible for the professional users (vapour pressure 6.5 Pa), but considering low concentration of ethylene glycol contained in the product (max. 0.47% in the concentrate and 0.024% in the in-use dilution) and downward application by low pressure spraying towards the ground, the exposure of professional users via inhalation is deemed to be very low. In addition to a SCOEL value, ethylene glycol has a skin notation and dermal exposure should therefore also be evaluated.

Based on the SCOEL of 52 mg/m3 (TWA 8 h), the AEL for an adult of 60 kg is calculated to be 52 mg/m3 x 1.25 m3/h x 8h ÷ 60 kg bw = 8.7 mg/kg bw

In the risk assessment for for monoethanolamine the AEL of 8.7 mg/kg bw is used as the reference value. The AEL of ethylene glycol for professionals is relatively high, showing low toxicity of the substance. To reach the AEL value a professional user of 60 kg needs to absorb the entire amount of ethylene glycol contained in 185 g of the concentrated product, which is an unlikely event. Moreover, when handling the concentrate, the use of gloves and a coverall is required due to classification of the product with H317, thereby further lowering the exposure via the dermal route. For exposure during application, the professional user even needs to absorb the entire amount of ethylene glycol in 3625g.

Altogether the exposure to ethylene glycol is not considered to cause adverse health effects to the professional users when using Biopren 50 LFL Fly larvicide concentrate in accordance

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to the use instructions. Due to its volatility ethylene glycol evaporates quickly after application from manure. Therefore secondary exposure to ethylene glycol is considered negligible.

Available toxicological data relating to a mixture

No studies using the formulation are available. The classification and labelling of the formulation has been prepared based on the calculation method described in Annex I of Regulation 1272/2008/EC.

Others

The Commission Delegated Regulation (EU) 2017/2100 specifying the scientific criteria for the determination of endocrine-disrupting properties (ED criteria) under Regulation (EU) No 528/2012 (BPR) establishes that the ED criteria become applicable by 7 June 2018 for biocides. No toxicological studies are available for the product Biopren 50 LFL FLY Larvicide cocnentrate. The product was not tested for potential endocrine disruption properties.

According to the CAR there is no indication that s-methoprene affects the endocrine system. To examine if any of the co-formulants contained in the product may possess ED properties, a screening was performed by examining whether the co-formulants are • Classified as CMR or PBT; • Identified as ED in the DG Santé’s Impact Assessment study on Screening of available evidence on chemical substances for the identification of endocrine disruptors; o Identified as ED in the EU list of potential endocrine disruptors; or o Listed in CoRAP linked to ED concerns. o included in ToxCast and EDSP21.

ED screening was performed by NL CA for all co-formulants, resulted in one potential alert. For one of the co-formulants (see onfidential annex for details) an ED concern has been raised by France and they propose to include this substance on the CoRAP list. CA NL cosiders that the ED assessment for this co-formulant does not need to be included in the ED assessment of the PAR and await the outcome of the discussions at EU level.

NL CA concludes that Biopren 50 LFL FLY Larvicide concentrate does not possess co- formulants with ED alerts for human health.

2.2.6.2 Exposure assessment

The active substance, S-methoprene and the product Biopren 50 LFL fly larvicide concentrateare manufactured and formulated in Hungary. No human health exposure scenarios have been assessed for the manufacture of the active substance or for the formulation of the product since production and formulation of the biocidal product is covered by other legislation.

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Potential exposure to S-methoprene from Biopren 50 LFL fly larvicide concentrate will occur through use of the product. Exposure assessment for use of this product is undertaken below. reparation has to be spread onto the manure’s surface either with low-pressure hand-held or backpack sprayer, or larger spraying tanks (e.g.50-100L) that may be moved e.g. with a trolley, at a dose of 230-240 ml of BIOPREN 50 LFL concentrate diluted to 5 litres with water /100 m 2. Slatted floor: it is necessary to repeat treatment after each removal of dung. Deep litter: on every new layer of litter and after every 8 - 10 cm increase the treatment has to be repeated with the product.

Inhalation exposure S-methoprene is not volatile and so the risk of inhalation exposure to S-methoprene for professional users during use is considered to be negligible during mixing and loading and post-application. However, during application by spraying aerosols are formed, this is taken into account for the exposure assessment.

Dermal exposure The main route of exposure is dermal exposure of users during manual mixing and loading, during application and during cleaning of equipment. After application, non-users are not likely to come into contact with ‘Biopren 50 LFL fly larvicide concentrate’ used in manure treatment. However as a worst case, secondary exposure calculation is also included.

Oral exposure ‘Biopren 50 LFL fly larvicide concentrate ’ is for professional use. Professional users are expected to follow good occupational hygiene where oral exposure will not occur. Therefore, the risk during use is considered to be negligible. Infant and children are not present in the animal houses. However children may visit animal farms. Therefore, this exposure scenario is taken into account for indirect exposure.

Identification of main paths of human exposure towards active substance(s) and substances of concern from its use in biocidal product

Summary table: relevant paths of human exposure Primary (direct) exposure Secondary (indirect) exposure Exposure Industri Profession Non- Industri Profession Gener Via path al use al use profession al use al use al food al use public Inhalation n.a. yes n.a. n.a. yes no n.a. Dermal n.a. yes n.a. n.a. yes yes n.a Oral n.a. no n.a. n.a. no yes n.a

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List of scenarios

Summary table: scenarios Scenario Scenario Primary or secondary exposure Exposed group number (e.g. Description of scenario (e.g. mixing/ professionals, loading) non- professionals, bystanders) 1. Mixing and Primary exposure Professional loading Based on HEEG Opinion 1, RISKOFDERM Dermal Model 2a Application Primary exposure Professional by low- Based on HEADhoc Recommendation 3, pressure RISKOFDERM, ART spraying 2b Application Primary exposure Professional by low- Based on US EPA Occupational Pesticide Handler pressure Unit Exposure Surrogate Reference Table spraying 3. Post- Primary exposure Professional application Cleaning of spraying equipment Based on HEAdhoc Recommendation 4 4 Workers Secondary exposure Worker secondary Based on HEEG Opinion 7 and HEAdhoc exposure Recommendation 12 5 Children Secondary exposure Children visiting Based on HEEG Opinion 7 and HEAdhoc animal Recommendation 12 farms 6 Dietary Secondary exposure General public exposure The general public could be exposed to residues via eating food from animal origin

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Industrial exposure

The active substance, S-methoprene and the product BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE are manufactured and formulated in Hungary. Production and formulation of the biocidal product is covered by other legislation. Industrial use of the product is not intended.

Professional exposure

Professional exposure refers to professional users such as a Pest Control Operator (PCO) and / or farmers but they are considered as professionals. In general the professional user is subject to national worker protection legislation and has residual risk controlled through control measures, which may include the use of personal protection equipment if required.

The professional user is expected to use gloves, however, the Tier 1 evaluation does not include any PPE.

Primary exposure to biocidal products occurs to the individual who directly uses /applies the product. Three main primary exposure scenarios are assessed for professional users: the mixing&loading phase, application and post-application.

During the mixing&loading phase (i.e. pouring the product concentrate into the spraying equipment and diluting it with water), only dermal exposure is relevant, as the active substance is not volatile and the formation of inhalable droplets is not expected.

During the application phase, primary exposure via skin contact or via inhalation may occur to the dilution. Professional users are expected to follow good occupational hygiene and thus contamination by ingestion is not expected. The oral route is not considered any further.

For the post-application phase (cleaning of the spraying equipment), only dermal exposure is considered relevant. Exposure may occur only to the diluted solution during this step. For further details see tables and calculations below.

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Scenario 1 Professional use, mixing and loading

For the treatment of fly larvae, 240 ml Biopren 50 LFL FLY Larvicide concentrate has to be diluted into 5 L working solution. This amount of working solution is sufficient for the treatment of 100 m 2 surface. The solution should be freshly prepared before application, proportionately to the area to be treated, based on this indicated dosage.

Mixing of the concentrate with water is not performed through stirring in the open equipment but only after the equipment has been closed, either by shaking (smaller sprayers) or through the automatic pumping of the device, so no exposure arises from this step. The only exposure is possible when the product is poured from its container into the device. The container of the product is designed in such a way that helps to minimize any accidental spills asd splashes when its content is poured. Exposure will be minimal.

Spraying is only performed with low-pressure sprayers (1-3 bar pressure). Hand-held or backpack sprayers (usually containing 5-20 L solution) are used; for larger areas the use of larger tanks (e.g. 50-100 L) is also possible which is then moved e.g. on a trolley.

For the task of spraying, a daily application time of 3 hours is considered relevant. The usual application rate of larvicide treatment is 0.3-0.5 L/min, this rate is appropriate to provide a uniform covering of the manure surfaces. With a 0.5 L/h rate, the user can treat 1800 m 2 surface. Among the housing areas relevant for cattle and pigs (the livestock buildings that may be treated with the product), the largest are the buildings of individually housed sows (460 m 2, based on the the OECD Emission Scenario Document for Insecticides for Stables and Manure Storage Systems). With this application time around 4 housings can be treated.

For the treatment specified above, max. 90 L working solution is required, which contains 4320 ml = 4.32 L Biopren 50 LFL Fly Larvicide concentrate. During the mixing&loading phase, this amount of product is poured into the spraying equipment.

Treatment may be performed with smaller or larger tanks – usually for smaller surfaces smaller tanks, and for larger surfaces larger tanks are used. If a tank of 5 L is used, 18 mixing&loading events would be required to prepare the dilution (18x5L = 90 L). In case a sprayer of e.g. 10 L is used, 9 mixing&loading events are necessary. For a tank of 100 L, only one loading event is required. Only the amount used for the treatment should be loaded in advance as the solution should always be prepared fresh. In any case, the product amount to be used will always be the same (4.32 L), and if smaller sprayers are used, the exposure time for loading is also proportionately shorter for each event but the overall loading time during the day will be the same. The final exposure will be the same in all cases.

The loading of the product concentrate into the device only takes a few seconds. The majority of the time when preparing the dilution is the task of adding water, but exposure to the product is no longer relevant in this phase. Mixing of the concentrate and the water always occurs in closed tanks as mentioned above. Pouring of 1L product does not take longer than 1 minute; for loading the total amount of 4.32 L product, 4.32 minutes will be required. Calculations have been therefore based on a total of 5 minutes for pouring all required product into the equipment.

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For the estimation of exposure during mixing&loading, the Biocides Human Health Exposure Methodology (2015) recommends as a first step to use HEEG Opinion 1 on the loading of products into vessels or systems in industrial scale. For liquid manual loading/pouring, HEEG Opinion 1 lists several available models; from these models the recommended choice for repeated loading (which is relevant for this product as larger quantities of working solution may be used during a day) is BEAT or RISKOFDERM Dermal model. Application of the product is also assessed with RISKOFDERM, so for a uniform approach RISKOFDERM is also used for the calculation of exposure during loading.

In HEEG Opinion 1, some default values for loading are shown for the RISKOFDERM Toolkit and RISKOFDERM Dermal model; the Excel version of RISKOFDERM Dermal model is also an annex to this guidance document. As specific parameters related to the use of the product are available, the Excel version of RISKOFDERM Dermal model can be configured the most to follow realistic conditions and is thus the most representative model for the current scenario.

The following parameters have been set in the RISKOFDERM Dermal Model:

Quality of ventilation related to the task: normal or good ventilation Frequency of skin contact with the contaminant: rare contact – it happens sometimes, but on average less than once per scenario. Exposure is only expected occssionally. Type of skin contact with the contaminant : more than light contact (splashes or drops fall onto the worker, part of the worker is in direct contact with the product) – in case exposure happens, it may be more than light, as a worst case approach Type of product handled: liquid Aerosols or splashes generated in the task: No (not generated) Level af automation of the task done by the worker: manual task Use rate of product: 1 L/min – it may be expected that 1 L product is poured in 1 minute into the equipment. For the remaining part of the loading process only water is poured which does not result in exposure of the worker. Percentile for exposure rate distribution to be assessed: 75 th percentile - to be in line with the RISKOFDERM assessment for application, as recommended by HEAdhoc Recommendation 3. Cumulative duration of scenario during a shift: 5 minutes (total)

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Description of Scenario 1: Professional user, mixing and loading product Professional user pouring product concentrate into spraying equipment and diluting it with water according to label instructions to create the working solution.

Only dermal exposure is included as inhalation exposure during mixing and loading is considered negligible (the active substance is non-volatile and no aerosols are formed during mixing and loading).

Based on RISKOFDERM Dermal model (Excel) Parameters Value Tier 1 (no PPE) active substance content in concentrate 5.26% (technical) dermal absorption (concentrate) 25% (default from EFSA guidance on dermal absorption, 2017) Inhalation absorption 100% User body weight 60 kg (based on HEAdhoc Recommendation 14 replacing HEEG Opinion No. 17, Default human factor values) Amount of product loaded 4.32 L (as calculated above) Dose for treatment 240 ml product/5L (label instructions, worst case) solution/100 m 2 surface Total loading time 5 min Body exposure 2.02 mg/min (HEEG Opinion 1; RISKOFDERM Dermal model, loading liquid default) Body exposure 3390 mg/min (HEEG Opinion 1; RISKOFDERM Dermal model, loading liquid default) Tier 2 (with Protection factor, gloves and coverall 90% PPE) (based on Biocides Human Health Exposure Methodology, 2015)

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The following exposure estimates have been found based on RISKOFDERM Dermal model with the above settings, without the use of personal protective equipment (Tier 1):

median percentile distribution Resulting exposure rate 2.78 µl/min 8.69 µl/min hands Exposure loading per shift 13.9 µl 43.4 µl hands

43.4 µl product concentrate is equivalent to 44.55 mg product (density = 1.0265 g/ml). This amount of Biopren 50 LFL FLY Larvicide concentrate contains 2.34 mg S-methoprene (technical). The Excel version of RISKOFDERM Dermal Model only estimates hand exposure but as a worst case body exposure may also be assumed. Exposure calculations have been based on the default value shown in HEEG Opinion 1 for RISKOFDERM Dermal Model, which is 2.02 mg/min product for body exposure. As the total duration is 5 minutes, the total exposure to the body is 10.1 mg product, which contains 0.531 mg active substance.

The total external dermal exposure is thus 2.34 + 0.531 = 2.87 mg S-methoprene from the loading phase.

Based on the default EFSA dermal absorption value of 25% (exposure to the concentrate) and a body weight of 60 kg, the systemic dose is 0.012 mg/kg bw/day , without personal protective equipment.

Professional users are always expected to wear personal protective equipment (PPE) when performing loading of the product, especially since the concentrate is classified as H317 which required the use of PPE (gloves and protective clothing). Tier 2 calculations are thus also presented below.

According to the Biocides Human Health Exposure Methodology, the protection factor of protective gloves and coverall is 90% for challenges by a liquid.

Based on these protection factors, external exposure is reduced to 0.234 mg active substance when gloves are worn. Body exposure is estimated to be 0.053 mg active substance when protection provided by clothing is accounted for. Total external dermal exposure is therefore 0.287 mg S-methoprene.

On the basis of the default EFSA dermal absorption value of 25% (exposure to the concentrate) and a body weight of 60 kg, the systemic dose when PPE (gloves and coverall) is worn is 0.0012 mg/kg bw/day .

eCA note: the applicant did provide tier 2 calculations assuming that gloves and a coverall are worn during mixing/loading activities. This was done because professional users are always expected to wear personal protective equipment (PPE) when performing loading of the product. Biopren 50 LFL FLY larvicide concentrate is classified as H317 and therefore PPE are required when handling the concentrate. However, the eCA notes that a tier 2 assessment is not required based on the systemic risk assessment (please refer to the risk characterisation for human health presented below).

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Scenario 2a: Professional user, Application by spraying (Based on HEAdhoc Recommendation no.3)

In HEAdhoc Recommendation No. 6 on the Methods and models to assess exposure to biocidal products in different product types, a recommendation is listed for the professional control of fly larvae in stables and cages by coarse, downward spraying. This guide recommends HEAdhoc Recommendation no. 3 - Spraying models for assessing exposure to insecticides for low pressure downward uses for assessing this scenario. Calculations have been therefore based on this guidance document (HEAdhoc 3).

In HEAdhoc Recommendation 3, it is proposed to use a combination of the exposure assessment models ART and RISKOFDERM for the estimation of inhalation and dermal exposure, respectively. Consequently, an assessment is presented below based on this approach.

As indicated above, spraying is only performed with low-pressure sprayers (1-3 bar pressure). Hand-held or backpack sprayers (usually containing 5-20 L solution) are used; for larger areas the use of larger tanks (e.g. 50-100 L) is also possible which is then moved e.g. on a trolley.

For the task of spraying, a daily application time of 3 hours is considered relevant. The usual application rate of larvicide treatment is 0.3-0.5 L/min, this rate is appropriate to provide a uniform covering of the manure surfaces. With a 0.5 L/min rate, the user can treat 1800 m 2 surface. Among the housing areas relevant for cattle and pigs (the livestock buildings that may be treated with the product) the largest are the buildings of individually housed sows (460 m 2, based on the the OECD Emission Scenario Document for Insecticides for Stables and Manure Storage Systems). With the specified application time about 4 housings can be treated.

For the treatment specified above, 90 L working solution is applied, which contains 4320 ml = 4,32 L Biopren 50 LFL Fly Larvicide concentrate. The following parameters have been set in the RISKOFDERM Dermal Model, as recommended by HEAdhoc Recommendation 3:

- indoor application - spraying downward - direction of airflow not clearly away from worker - no segregation of the worker from the source - source less than 1 meter from the worker - liquid not highly volatile - application rate 0.5 L/min - percentile 75 th - exposure duration: 180 minutes

The settings used for the ART modelling are the following: - indoor application - spraying downward - no segregation of the worker from the source

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- source less than 1 meter from the worker (near field) - liquid non-volatile - application rate 0.3-3 L/min - No restriction on general ventilation characteristics - vapour pressure 0.00108 Pa - exposure duration: 180 min - liquid weight fraction: 0.00015

Description of Scenario 2a: Professional user, application by spraying According to HEAdhoc Recommendation 3, Spraying models for assessing exposure to insecticides for low pressure downward uses: based on RISKOFDERM Dermal model (Excel) and ART.

Professional user spraying working solution with the use of hand-held or backpack spraying equipment, for the treatment of fly larvae in livestock buildings.

Dermal and inhalation exposure is possible. Oral exposure is not considered relevant for professional users. Parameters Value Tier 1 (no PPE) active substance content in working 0.2592% solution (technical) dermal absorption (dilution) 50% (default water-based products, EFSA 2017) Inhalation absorption 100% User body weight 60 kg (based on HEAdhoc Recommendation 14 replacing HEEG Opinion No. 17, Default human factor values)

Application time 180 min Application rate 0.5 L/min Dose for treatment 240 ml product/5L (label instructions, worst case) solution/100 m 2 surface Tier 2 (with Protection factor, gloves and coverall 90% PPE) (based on Biocides Human Health Exposure Methodology, 2015)

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Dermal exposure, RISKOFDERM:

Based on the model, the resulting exposure to the working solution is as follows: median percentile distribution (75 th ) Resulting exposure rate 8.72 µl/min 29.2 µl/min hands Resulting exposure rate 33.6 µl/min 112 µl/min body Exposure loading per shift 1570 µl 5260 µl hands Exposure loading per shift 6040 µl 20200 µl body Total dermal exposure per 7610 µl/day 25460 µl/day shift

The concentrate product contains 5.26% S-metoprene. The dilution rate is 240 ml product to 5 L working solution, resulting in 0.2592% active substance in the working solution. Consequently, the estimated daily dermal exposure of 25460 µl (25460 mg – as the dilution is mainly water, its density may be considered 1) equals to an external dermal exposure of 65.98 mg active substance.

With the dermal absorption value of 50% and a user body weight of 60 kg, the resulting internal systemic exposure is 0.550 mg/kg bw/day .

For the above calculations the use of PPE was not considered (Tier 1). Nevertheless professional users will always wear personal protective equipment for the use of the product. Tier 2 values are therefore also presented below.

According to the Biocides Human Health Exposure Methodology, the protection factor of protective gloves and coverall is 90% for challenges by a liquid.

Based on these protection factors, external total dermal (hand and body) exposure is reduced to 6.60 mg active substance when gloves and coverall are worn.

On the basis of a dermal absorption value of 50% and a body weight of 60 kg, the systemic dose from dermal exposure when PPE is worn is 0.055 mg/kg bw/day .

Inhalation exposure, ART:

As a result of ART inhalation modelling, it was concluded that the estimated inhalation exposure was 0.0006 mg/m³ active ingredient (75 th percentile value).

Based on HEAdhoc Recommendation no. 14 revising HEEG Opinion 17 or ECHA’s Biocides Human Health Exposure Methodology, the default adult inhalation rate is 1.25 m 3/h . With an exposure duration of 180 min , a total of 3.75 m 3 air is inhaled. The resulting exposure

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is 0.00225 mg active substance per day. With a user body weight of 60 kg , the resulting systemic exposure is 3.75x 10 -5 mg/kg bw/d .

Scenario 2b: Professional user, Application by spraying (Based on US EPA Occupational Pesticide Handler Unit Exposure Surrogate Reference Table)

eCA note: please note that the applicant provided an additional model to determine the exposure by spraying in addition to the harmonized models described in HEAdhoc Recommendation No. 6. The applicant indicated the US EPA guide of 2018 shows more relevant, measured values for the intended pattern. HEAdhoc 3 is more a general spraying model therefore the EPA values may be even more representative for the current use. Therefore it is preferred to include these results in the PAR as well as a comparison.

However, the eCA considers the US EPA PHED model not appropriate, as it is included in the text of the guidance: There is only ONE study in PHED that estimates exposure for this use scenario (#1024). The data were monitored in greenhouses; applications to floor, bench, and overhead plants (nozzle pressure not reported).

And

Low Confidence due to inadequate replicate number.

However, as the applicant wishes to keep an assessment using this model in the PAR the eCA did not remove this scenario 2b, but please note that scenario 2b was not evaluated by the eCA since the exposure assessment was based on scenario 2a using harmonised models according to HEAdhoc recommendation no. 3.

Furthermore, during mutual recognition the dermal absorption value for the diluted product was changed to 50%. This is not adapted in scenario 2b.

As a comparison, a different model is also presented for the assessment for fly treatment. The US EPA Occupational Pesticide Handler Unit Exposure Surrogate Reference Table (June, 2018) presents the current recommended unit exposures for standard Agency occupational pesticide handler exposure scenarios. These values are derived from a number of sources, including the Pesticide Handler Exposure Database (PHED), the Outdoor Residential Exposure Task Force (ORETF) the Agricultural Handler Exposure Task Force (AHETF), Exit or other available registrant-submitted exposure monitoring studies.

This EPA guide contains reference values that are specifically recommended for livestock houses therefore may be even more relevant than the general spraying models available in the EU. Therefore the results obtained based on these values are considered relevant and representative for the product.

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Three scenarios are present in the US EPA guide that may be relevant for the product: 1. Mixer/loader/Applicator, Manually-pressurized handwand; 2. Mixer/Loader/Applicator, Backpack sprayer and 3. Mixer/Loader/Applicator, Mechanically-pressurized handgun sprayer. From these scenarios, the Manually-pressurized handwand scenario has the highest Tier 1 values but the second scenario, the Backpack sprayer model has the higest recommended Tier 2 values. PPE will be mandatory for the use of the product therefore as a worst case approach the Backpack sprayer scenario is presented below.

Description of Scenario 2b: Professional user, application by spraying According to US EPA Occupational Pesticide Handler Unit Exposure Surrogate Reference Table (June, 2018) - Mixer/Loader/Applicator, Backpack sprayer

Professional user spraying working solution with the use of hand-held or backpack spraying equipment, for the treatment of fly larvae in livestock buildings.

Dermal and inhalation exposure is possible. Oral exposure is not considered relevant for professional users. Parameters Value Tier 1 (no PPE) active substance content in concentrate 5.26% (technical) dermal absorption (dilution) 35% (from oral absorption value – see explanation above) Inhalation absorption 100% User body weight 60 kg (based on HEAdhoc Recommendation 14 replacing HEEG Opinion No. 17, Default human factor values)

Applied amount of working solution 90 L = 90 kg Dose for treatment 240 ml product/5L (label instructions, worst case) solution/100 m 2 surface Dermal exposure, no gloves 2510 μg/lb ai. = 5.53 mg/kg (based on US EPA guide) handled ai. Inhalation exposure (no respirator) 30 μg/lb ai. = 0.066 mg/kg (based on US EPA guide) handled ai. Tier 2 (with Dermal exposure, single layer gloves 2500 μg/lb ai. = 5.51 mg/kg PPE) (based on US EPA guide) handled ai.

The scenario presented above contains both mixing&loading and application phases.

Based on the values shown also in the previous scenario, the applicator sprays 90 L = 90 kg (density may be considered as 1) working solution. This amount of solution contains 0.233 kg S-methoprene. This means that external dermal exposure will be 1.291 mg active

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ingredient. With a dermal absorption of 35% and a body weight of 60 kg, the internal dose is 0.00753 mg/kg bw/day without PPE.

When calculated with the value measured for single layer goves, the external exposure is 1.286 mg active ingredient. With a dermal absorption of 35% and a body weight of 60 kg, internal dose is 0.00749 mg/kg bw/day with the use of single layer gloves.

The inhaled amount is 0.015 mg S-methoprene. With a body weight of 60 kg, the internal dose from inhalation is 0.00026 mg/kg bw/day .

Total internal exposure in Tier 1 (no PPE) is therefore 0.00779 mg/kg bw/day . When single layer gloves are also used, the exposure is 0.00776 mg/kg bw/day (Tier2).

Scenario 3: Professional user, Post-application – cleaning equipment after treatment

Description of Scenario 3: Post application step, cleaning of spray equipment Professional user cleaning the spray equipment which contains any remaining dilution. This step is performed once at the end of the working day.

240 ml product was diluted to 5 L working solution. Active substance content in product: 5.26%. Active substance content in di lution: 0.259%. Dilution density may be considered as 1.

Estimated on the basis of HEAdhoc Recommendation no. 4. Parameters Value Tier 1 (no PPE) body exposure (HEAdhoc 4) 19.28 μl/min hand exposure (HEAdhoc 4) 35.87 μl/min duration of cleaning (HEAdhoc 4) 20 min dermal absorption (dilution) 50 % (default water-based products, EFSA 2017) User body weight 60 kg (based on HEAdhoc Recommendation 14 replacing HEEG Opinion No. 17, Default human factor values) Tier 2 (with Protection factor, gloves and coverall 90% PPE) (based on Biocides Human Health Exposure Methodology, 2015)

A scenario is included for the cleaning of the spray equipment. Calculations are based on Recommendation no. 4 of the BPC Ad hoc Working Group on Human Exposure (HEAdhoc) (2014).

This recommendation may be considered as an absolute worse case for the current scenario as HEAdhoc recommendation 4 concerns the cleaning of spray equipment in antifouling use

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

(paint spray equipment). The working solution of Biopren 50 LFL FLY Larvicide concentrate is not a viscous solution, it may be cleaned very easily, thus the total time required for the cleaning of the equipment should not take more than 10 minutes. The default value suggested in the Recommendation was 20 minutes, this value was used in the calculations as an absolute worst case, which may be considered as an overestimation of actual expected exposure. Based on the HEAdhoc Recommendation 4 and realistic post-application conditions, only dermal exposure was considered relevant for this scenario.

On the basis of the above-listed parameters, the body exposure to the working solution (which is present in the spraying device and exposure may occur during cleaning), is 385.6mg . Hand exposure is 717.4mg . The density of the solution is 1 (it is mainly water). Thus, the total external dermal exposure is 1103 mg working solution/day .

The active substance content of the concentrate is 5.26% and the dilution is prepared by diluting 240 ml product to 5 L working solution. Active substance content in the dilution is 0.25917%. Consequently, the total external dermal exposure is 2.859 mg active substance/day.

Dermal absorption was assumed to be 50% as a worst case based on the default water- based products (EFSA 2017), however realistic absorption values are expected to be much lower than this amount. As no specific study with Biopren 50 LFL FLY Larvicide concentrate are available, this default was used as an absolute worst case approach. The absorbed amount is therefore 1.43 mg active substance /day .

With a body weight of 60 kg , the internal dose from this scenario is 0.024 mg active substance/kg bw/day , when no personal protective equipment is used (Tier 1).

Professional users are always expected to wear protective equipment when performing the cleaning of the spraying equipment. Tier 2 values are therefore also presented below.

According to the Biocides Human Health Exposure Methodology, the protection factor of protective gloves and coverall is 90% for challenges by a liquid.

Based on these protection factors, the total external dermal exposure (hand and body exposure) to the working solution is reduced to 0.286 mg S-methoprene.

On the basis of a dermal absorption value of 50% and a body weight of 60 kg, the systemic dose from dermal exposure when PPE is worn is 0.0024 mg/kg bw/day .

Scenario 4: Professional worker, secondary exposure

The workers in livestock buildings may accidentally touch the treated areas. The product is applied for fly larvae treatment on manure; treated manure is usually not touched by hand and gloves are worn for such situations. In realistic use conditions secondary exposure through treated surfaces does not occur or is negligible.

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Description of Scenario [4] professional worker, s econdary exposure Parameters Value

Tier 1 Dose 2.46 g/m 2 product*

fraction of active substance on indoor surface 100% (worst case)** that is available for transfer

Concentration of the active substance 0.2592%

Dermal absorption 50%

Surface area (both palms) HEAdhoc 410 cm2/event Recommendation No. 14

Body weight (HEAdhoc Recommendation No. 60 kg 14) * application rate = 246,36 g product (240 ml with a density of 1.0265 g/ml) to be used on 100m 2 = 2.46 g/m 2 ** The treated surface that is touched may be wet. As there are no appropriate parameter available the worst case (100% transfer) is considered.

Dermal exposure is calculated as following: Dose x fraction available for transfer x concentration of the a.s. x dermal absorption x surface area (both palms) /body weight = 2.46 g/m 2 product x 100% transfer x 0.2592% a.i. x 50% dermal absorption x 410 cm2 /60 kg bw = 0.022 mg/kg bw

Calculations for Scenarios 1-4

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Summary table: estimated exposure from professional uses Exposure Tier/PPE Estimated Estimated Estimated Estimated scenario inhalation dermal oral uptake total uptake uptake uptake Scenario 1, Tier 1/ negligible 0.012 mg/kg n.a. 0.012 mg/kg Professional no PPE bw/day bw/day user, loading Scenario 1, Tier 2/ negligible 0.0012 mg/kg n.a. 0.0012 mg/kg Professional with PPE bw/day bw/day user, loading Scenario 2a Tier 1/ 3.75x 10 -5 0.550 mg/kg n.a. 0.550 mg/kg Professional no PPE mg/kg bw/day bw/day bw/day user, Application by spraying (based on HEAdhoc 3) Scenario 2a Tier 2/ 3.75x 10 -5 0.055 mg/kg n.a. 0.055 mg/kg Professional with PPE mg/kg bw/day bw/day bw/day user, Application by spraying (based on HEAdhoc 3) Scenario 3, Tier 1/ negligible 0.024 mg/kg n.a. 0.024 mg/kg Professional no PPE bw/day bw/day user, Post- application Scenario 3, Tier 2/ negligible 0.0024 mg/kg n.a. 0.0024 mg/kg Professional with PPE bw/day bw/day user, Post- application Scenario 4, Tier 1/ negligible 0.022 mg/kg n.a. 0.022 mg/kg Professional no PPE bw/day bw/day worker, Secondary exposure

Further information and considerations on scenarios [1-4]

Exposure assessments have been based on the Biocides Human Health Exposure Methodology (2015), the relevant HEEG Opinions and HEAdhoc Recommendations and EFSA guidance on dermal absorption (2017) (for detailed references see above).

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

During the product life cycle human exposure may occur at time of loading of the product, application and post-application. Professional users are assumed to wear protective gloves and coverall when handling the products. As a first step, exposure has been estimated without the use of any protective equipment (Tier 1).

It is assumed that 100% of inhalation exposure is absorbed. For dermal absorption of the concentrate, the default value from the EFSA guide on dermal absorption has been used, for the worst-case estimation of the absorption of the dilution the oral absorption value of 35% has been used; please see above for a detailed explanation. Operator body weight is assumed to be 60 kg.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Combined scenarios

It is realistic for professional users that the same person will be performing the mixing&loading, application and post-application tasks, therefore the assessment of combined exposure from these scenarios is relevant. The combined results of these tasks can be found in the table below.

Scenario 4 is not considered relevant for combined exposure. Usually the pest control operator and the worker handling manure in livestock buildings is a different person and treatment of surfaces and handling of treated manure will not occur on the same day. A combination of these doses is therefore not relevant. Scenario 4 is considered separately in the risk assessment.

Summary table: combined systemic exposure from professional uses Scenarios Estimated Estimated dermal Estimated oral Estimated total combined inhalation uptake uptake uptake uptake Scenarios 3.75x 10 -5 0.586 mg/kg bw/day n.a. 0.586 mg/kg [1+2a+3] mg/kg bw/day bw/day Tier 1, no PPE Scenarios 3.75x 10 -5 0.059 mg/kg bw/day n.a. 0.059 mg/kg [1+2a+3] mg/kg bw/day bw/day Tier 2 (gloves and coverall)

Non-professional exposure

Not applicable. BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE is not intended for non- professional use.

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Exposure of the general public

Scenario 5: Secondary exposure, children visiting animal farms

In general infants and children are not present in the animal houses. The secondary exposure of children is not a realistic scenario as children will not have access to treated manure of livestock buildings; however as a worst case approach calculations have been presented. Palms of a child may be exposed dermally touching the treated floor. Hand-to- mouth exposure is a behaviour normally included for the exposure of children, however, as the product is spread directly on the manure, therefore there is no direct contact without touching the manure. Oral exposure due to hand-to mouth behaviour by touching treated manure is not considered a realistic exposure. Subsequently, only dermal exposure is considered.

A child may be exposed by touching the treated area in livestock buildings.

Description of Scenario [5] secondary exposure, children visiting animal farms Parameters Value

Tier 1 Dose 2.46 g/m 2 product*

fraction of active substance on indoor surface 100% (worst case)** that is available for transfer

Concentration of the active substance 0.2592%

Dermal absorption 50%

Surface area (both palms) HEAdhoc 115.2 cm2/event Recommendation No. 14

Body weight (HEAdhoc Recommendation No. 10 kg 14) * application rate = 246,36 g product (240 ml with a density of 1.0265 g/ml) to be used on 100m 2 = 2.46 g/m 2 ** The treated surface that is touched may be wet. As there are no appropriate parameter available the worst case (100% transfer) is considered.

Dermal exposure is calculated as following:Dose x fraction available for transfer x concentration of the a.s. x dermal absorption x surface area (both palms) /body weight = 2.46 g/m 2 product x 100% transfer x 0.2592% a.i. x 50% dermal absorption x 115.2 cm2 /10 kg bw = 0.0367 mg/kg bw

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Calculations for Scenario 5

Summary table: systemic exposure of general public Exposure Tier/PPE Estimated Estimated Estimated Estimated total scenario inhalation dermal oral uptake uptake uptake uptake Scenario Tier 1 (no - 0.0367 n.r. 0.0367 mg/kg 5, PPE) mg/kg bw/d children bw/day indirect exposure

Further information and considerations on scenario [5]

No further information is available.

Combined scenarios

Only one secondary exposure scenario is relevant therefore combined scenarios are not applicable for secondary exposure.

Monitoring data

No monitoring data is available.

Dietary exposure

During the mixing & loading and the application scenarios no food, drinking water are exposed to the product. Therefore as no food is contaminated no Dietary Risk Assesment (DRA) is required. However, as animal exposure cannot be excluded exposure via eating or drinking products from animal origin needs to be evaluated. This is considered in the paragraph Estimating Livestock Exposure to Active Substances used in Biocidal Products below.

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Information of non-biocidal use of the active substance

In Europe S-methoprene is authorized for biocidal and veterinary use. In Australia it is also used for grain treatment.

Residue definitions [Summary table of other (non-biocidal) uses Sector of Intended use Reference value(s) 2 use 1 1. Biocide, EU Not establised 2. Veterinary, Spot-on products Not establised EU 3. Agriculture, Grain treatment Australia 4. No approved As the Maximum Residue for Plant Levels in animal products are Protection 0.05* mg/kg based on the Products lower limit of analytical determination (in some of the products 0.1 mg/kg as edible offal of swine, bovine, sheep or 0.2 mg/kg as fat free of lean meat of swine, bovine, sheep (see Regulation 899/2012/EC for PPP)

1 e.g. plant protection products, veterinary use, food or feed additives 2 e.g. MRLs. Use footnotes for references .

Estimating Livestock Exposure to Active Substances used in Biocidal Products

Livestock are not present when the litter or manure is treated. The only possible exposure is via secondary exposure. With Biopren 50 LFL Fly Larvicide concentrate only treatment of piggeries, sheepfolds and cattle farms is permitted. Therefore an exposure screening was performed for these livestock based on the DRAWG Draft proposal guidance on Estimating Livestock Exposure to Biocidal Active Substances (2010) included in BPR guidance (2017, version 4).

Based on the above-mentioned guidance, the following screening step may be applied:

Exposure = AR x AT : N : BW

Where: AR: application rate = 129.59 mg a.i./m2 (calculated from the dose of 240 ml product/100 m2, with a product density of 1.0265 and an a.i. content of 5.26%) AT: area treated N: number of animals per stable body weight of animal

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

In the screening calculations above, an absolute worst case value is estimated where all applied product on the surface is accessible to the animals. Based on the guidance, uptake of the entire amount of applied product is assumed in this step regardless of the route of exposure. The calculated dose will therefore overestimate actual exposure.

For treated areas the values from the ESD for Insecticides for Stables and Manure Storage Systems were applied. The area of walls is not relevant for larvicide treatment therefore the default surfaces intended for larvicides were used. Numbers of animals per stable and body weight values originate from Appendix 6-1 of the Guidance on BPR: Volume III Parts B+C. As no information is included for animal housing for sheep/lamb, these are not included in the sceeening below.

Please find below the results of the screening assessment for the relevant livestock:

Livestock floor area per Number of Body weight Exposure stable (m 2) animals/stable (screening) (mg/kg bw/ day) Dairy cattle 1170 100 650 kg 2.33 Beef cattle 370 125 500 kg 0.77 Veal calves 160 80 200 1.30 Sows (group 710 132 260 2.68 housing, worst case) Fattening pigs 710 400 100 1.94

To derive an AEL for animals, an assessment factor of 5 (for human 100 is used) can be applied according to the EFSA guidance on birds and mammals (2009). From the NOAEL 21.7 mg/kg bw, by applying correction for oral absorption (0.35) and the assessment factor of 5, the AEL of 1.52 mg/kg bw can be derived for animals. The screening value for dairy cattle, fattening pigs and group housed sows exceed the AEL, therefore a realistic worst- case estimate is presented below for all livestocks:

Oral exposure through licking surfaces: The exposure may be calculated as: AR x TS x LD : BW

Where: AR: application rate = 129.59 mg a.i./m 2 as calculated above TS: tongue surface area LD: licks per day BW: body weight

Oral exposure through uptake of contaminated feed in through:

The exposure may be calculated as: AR x ES : BW

Where:

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

AR: application rate = 129.59 mg a.i./m 2 as calculated above ES: exposed feed surface BW: body weight

Dermal exposure through rubbing against surfaces :

The exposure may be calculated as: AR x BS : BW

Where: AR: application rate = 129.59 mg a.i./m 2 as calculated above BS: body surface in contact with surface BW: body weight

Tier 1 calculations are based on 100% oral and dermal absorption.

It should be noted that according to the Guidance on BPR, Volume III Parts B+C, for a treatment of the floor, an emission factor of 0.85 can be chosen and for oral exposure through uptake of contaminated feed by spraying on the through an emission factor of 0.11 could be used (fraction emitted to the treated surface during surface treatment by spraying). However, Biopren 50 LFL Fly Larvicide concentrate is intended to be used at a low application rate of 0.3-0.5 L/min as a more uniform, even treatment of the manure should be achieved instead of a quick treatment with larger volumes. This will result in a more targeted application and better effectiveness.It was therefore assumed in the calculations made that all of the product would reach the targeted floor and a difference was not made between the amount that would reach the floor and the amount that could be present on the walls.

Default values are based on Guidance on BPR: Volume III Parts B+C.

Defaults and results, Tier 1:

parameter Beef Dairy Veal Fattening Sows Sheep Lamb (see above) cattle cattle calves pigs TS (m2) n.a. n.a. 0.008 0.008 0.008 0.008 0.008 LD (no.) n.a. n.a. 10 10 10 10 10 BW (kg) 500 650 200 100 260 75 40 ES (m2) 0.7 2.9 0.5 0.4 1.1 n.a. n.a. BS (m2) 1.44 1.68 0.87 0.45 0.84 n.a. n.a. Oral exposure licking n.a. n.a. 0.0518 0.1037 0.0399 0.1382 0.2592 (mg/kg bw/d) Oral exposure contaminated 0.020 0.064 0.0356 0.0570 n.a. n.a. n.a. feed (mg/kg bw/d) Dermal exposure surface 0.373 0.335 n.a. 0.5831 0.4187 n.a. n.a. rubbing (mg/kg bw/d) Total 0.393 0.399 0.651 0.7438 0.4585 0.1382 0.2592 exposure

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

(mg/kg bw/d)

Inhalation exposure is not relevant for livestock as the animals are not present during treatment and the active substance is not volatile thus evaporation will not be relevant. Based on the BPR guidance, exposure from consumption of dead flies should not be included for beef and dairy cattle, sheep and pigs.

The exposure value represents an absolute worst-case scenario as all the applied amount was considered accessible to the animals which is a clear overestimation, furthermore reduction due to dermal and oral absorption was also not taken into account. As all calculated worst-case exposure values are below the AEL level specified for livestock, risk to livestock is considered acceptable and no further refinement e.g. using a factor of 50% for dermal and oral absorption, is proposed.

For consumer exposure , the standard food basket as defined in Commission Regulation (EU) 2018/782 is considered. According to the standard food basket the theoretical maximum daily intake for animal origin product for an adult is: 2.0 kg for mammals (0.3 kg muscle, 0.05 kg fat, 0.1 kg liver, 0.05 kg kidney 1.5 kg milk). Poultry is not relevant as the treatment of poultry housings is not possible with the product.

The theoretical maximum consumption of the substance is therefore calculated as follows (using Tier 1 calculations (100% oral and dermal absorption): 0.743 mg/kg bw (based on fattening pigs, as calculated above, representing the worst- case value) x 2.0 kg x 50% oral absorption = 0.743 mg/person = 0.012 mg/kg bw/day based on a 60 kg person.

For S-methoprene, no ADI is set in EU. The AEL long-term was derived from the NOAEL of 21.7 mg/kg bw/day from the 2-year rat study, using an assessment factor of 100 and corrected for oral absorption, resulting in a value of 0.076 mg/kg bw/day. For the derivation of an ADI, the NOAEL of 21.7 mg/kg bw/d from the 2-yr rat study is applicable and including an assessment factor of 100, a value of 0.217 mg/kg bw/day is derived.

The maximal theoretical consumption of 0.012 mg/kg bw/d is 5.7% of the derived ADI, therefore the risk is considered acceptable.

Conclusion

Based on the calculation of the exposure of livestock it is expected that the product BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE does not pose a concern for livestock and for consumers. The low toxicity of S-methoprene (acute as well as chronic toxicity) and the result of the toxicokinetic study (rapid elimination from the body, no accumulation in the body)further supports this conclusion.

However, as the trigger value of 0.004 mg/kg bw/d is exceeded for the representative animal species the issue of potential residues in food of animal origin must be further addressed

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

and this will be included in section 2.2.6.3 Risk characterision for Human Health when discussing MRLs.

Estimating residues in edible crops via fertilizer (scenario 6b)

As manure is commonly spread onto agricultural land as fertilizer, biocidal residues contained in manure may be transferred into crops grown on this land. For this, the concentration in plant roots due to the absorption of the s-methoprene via roots is calculated, based on the model used for environmental risk assessment for REACH. According to the EUSES background document,

Fwater plant : volume fraction of water in plant tissue [m3.m-3] Flipidplant: volume fraction of lipids in plant tissue [m3.m-3] Kow: octanol-water partition coefficient [m3.m-3] b: Correction for differences between plant lipids and octanol [-] Kplant-water : partition coeff. between plant tissue and water [m3.m-3]

Kplant-water: = 0.65 + 0.01 x 6.34 L/kgx0.95 = 0.71 m3/m3

Kplant-water partition coeff. between plant tissue and water [m3.m-3] Cagric,porew: in pore water of agricultural soil [kgc.m-3] (PEC value in groundwater) RHOplant: bulk density of plant tissue (wet weight) [kgwwt.m-3] Croot : concentration in root tissue of plant [kgc.kgwwt-1]

Croot: = (0.71 x 5.17 E-7 kg/m3)/700 = 5.24 E-10 kg a.s./kgwwt = 5.24E-4 mg/ kgwwt = 0.00052 mg/ kgwwt

The half-life of S-methoprene in the soil is 1.55 days, and the a.s. will rapidly decompose in field. Also, the instruction not to use treated manure earlier than 2 months after treatment is included. The decomposition of the a.s. during this two months in manure is not taken into account in the calculation for PECground water. The calculated concentration represents therefore the theoretical worst case and in practice the concentration should be much lower

Conclusion Based on the CA document for an interim approach for the establishment of MRL (CA- March17-Doc.7.6.c-final), the question of residues should be further explored only when active substances can lead to measurable residue levels in food (sec 2.2). The calculated S- methoprene level of 0.00052 mg/kg bw is below the default MRL of 0.01 mg/kg, which is set at the LOD, thus not measurable.

In addition, MRLs for S-methoprene are set to be 0.02-5 mg/kg for different food categories for both plants and animal origin, based on EU No 396/2005.

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Altogether, S-methoprene concentration in crop is considered not to be measurable, and is below the MRLs. Therefore no adverse effects on consumers via dietary exposure to food of plant origin of land fertilized with treated manure is expected.

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Estimating transfer of biocidal active substances into foods as a result of professional and/or industrial application(s)

No EU guideline is available for estimating transfer of biocidal active substances into foods as a result of professional application. However it is assumed that no food contamination would occur during the use of the product. BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE is used for manure treatment in animal stables and manure pits. Therefore the product is not a disinfectant, not used in residential homes, for drinking water disinfection and not an in-can preservative where food contamination is expected.

Estimating transfer of biocidal active substances into foods as a result of non- professional use

Not applicable. BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE is not intended for use by non-professional users. Dietary exposure by consuming animal products is considered in the section “Estimating Livestock Exposure to Active Substances used in Biocidal Products” above. Further no food contamination would occur during the use of the product. BIOPREN 50 LFL FLY CONCENTRATE is used for manure treatment in animal stables and manure pits.

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Exposure associated with production, formulation and disposal of the biocidal product

The active substance S-methoprene is manufactured in a closed system which is described in the confidential annex of the dossier supporting inclusion decision of S-methoprene. Full PPE is required (Gloves, coverall, face-shield and respirator) during filling and maintenance. No cleaning of the apparatus occurs since only S-methoprene is produced in the system. The only operator contact with the active ingredient is during sampling for quality.

Exposure during formulation of the product BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE is expected to be minimal due to operating in a closed system with a ventilation system of the premises. Measurement and mixing of components is automated and controlled by computer. During the production every worker must wear protective glasses, plastic gloves, mask and overall.

Therefore no hazard identified during manufacturing, and no risk assessment is needed.

There is no guide for calculation of the exposure associated with waste disposal. However based on the toxicological profile and fast photo- and biodegradation of S-methoprene in the environment no significant exposure is expected during the wate disposal of the product.

Aggregated exposure

No metholodology has been developed yet. S-methoprene is also authorized according to the veterinary regulation however S- methoprene mainly used in biocide products in the EU. These products are used in different areas, therefore there is no overlapping use of the products. Therefore no aggregated exposure estimation required

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Summary of exposure assessment

Scenarios and values to be used in risk assessment Scenario Exposed group Tier/PPE Estimated total number (e.g. professionals, non- uptake professionals, bystanders) (mg/kg bw) 1.Mixing&loadi Professional user Tier 1, no PPE 0.0120 mg/kg ng bw/day 1. Professional user Tier 2, with PPE 0.0012 mg/kg Mixing&loading (gloves, coverall) bw/day Scenario 2a Professional user Tier 1, no PPE 0.550 mg/kg Professional bw/day user, Application by spraying (based on HEAdhoc 3) Scenario 2a Professional user Tier 2, with PPE 0.055 mg/kg Professional (gloves, coverall) bw/day user, Application by spraying (based on HEAdhoc 3) 3. Post- Professional user Tier 1, no PPE 0.024 mg/kg application – bw/day cleaning of equipment 3. Post- Professional user Tier 2, with PPE 0.0024 mg/kg application – (gloves, coverall) bw/day cleaning of equipment 4. Workers Workers, secondary exposure Tier 1, no PPE 0.022 mg/kg accidentally bw/day touching treated manure 5. Children Children, secondary exposure Tier 1, no PPE 0.0367 mg/kg visiting farms bw/day

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2.2.6.3 Risk characterisation for human health

Reference values to be used in Risk Characterisation Reference Study NOAEL AF 1 Correction for Value (LOAEL) oral absorption (mg/kg (mg/kg bw/day) bw/day) AELshort- developmental 100 100 0.35 0.35 term rabbit study AELmedium- dog 90-day 100 100 0.35 0.35 term repeat oral AELlong-term combined 21.7 100 0.35 0.076 chronic toxicity and carcinogenicity ARfD Not -- established ADI Not -- established

The ARfD was not derived during the discussion phase of the active substance.

AELmedium-term: In the dog 90-day repeat oral dose study the NOAEL value of 100mg/kg bw/day is based on clinical signs and increased liver weight in both sexes and raised ALP in females at the mid- dose level assessed of 300mg/kg bw/day. At the highest dose level assessed the effects noted include gastrointestinal signs, increased liver weight, raised ALP levels and also zonal vacuolisation of hepatocytes. All of these effects were noted in both sexes at this dose level. This information indicates a clear dose response relationship and the effects noted including the vacuolisation of hepatocytes, which may be due to fatty or fluid balance change, may be indicative of liver toxicity. Accordingly from the results achieved, the NOAEL value of 100mg/kg bw/day obtained will be taken forward to the risk characterisation for medium- term repeated exposure and was used to establish a systemic AEL medium-term reference value of: AELmedium-term 0.35 mg/kg bw/day.

AELlong-term In the combined chronic toxicity and carcinogenicity study conducted in rat the NOAEL value of 21.7-mg/kg bw/day is based on evidence of liver toxicity such as increased incidence of hepatic lesions (bile-duct proliferation and portal lymphocyte infiltration) in males and increased absolute and relative weights of the liver in females obtained at the highest dose assessed which is the equivalent of 108 mg/kg bw/day S-methoprene. The value of 21.7mg/kg bw/day S-methoprene is taken forward to the risk characterisation for long-term repeated exposure and was used to establish a systemic AEL long-term reference value of: AELlong-term 0.076 mg/kg bw/day.

AELacute: In the developmental rabbit gavage study severe maternal toxicity (including mortalities and abortion) was accompanied by significant foetolethality and foetotoxicity at the high dose of 1000 mg/kg bw/day. At the next dose level assessed, 100-mg/kg bw/day, no effects were observed. The top dose is considered to be inappropriately high and the mid-range dose provides an NOEL value. However, this is used to establish a systemic AEL acute reference value. It is recognised the value used of 100mg/kg bw/day may be overly

NL Biopren 50 LFL FLY Larvicide concentrate PT 18 conservative but considering the inadequate dosing in the rabbit developmental study the value is brought forward to the risk characterisation for acute exposure and was used to establish systemic AEL acute reference value of: AELacute 0.35 mg/kg bw/day.

Maximum residue limits or equivalent

In the Approval Regulation of S-methoprene PT18 (in force on 1-9-2015) is described that for products that may lead to residues in food or feed, the need to set new or to amend existing maximum residue levels (MRLs) in accordance with Regulation (EC) No 470/2009 of the European Parliament and of the Council or Regulation (EC) No 396/2005 of the European Parliament and of the Council shall be verified, and any appropriate risk mitigation measures shall be taken to ensure that the applicable MRLs are not exceeded.

For the purpose of dietary risk assessment, and to establish the need to derive a MRL, and in absence of an EU harmonised ADI, an ADI was derived based on the information included in the S-Methoprene dossier. The AEL long-term was derived from the NOAEL of 21.7 mg/kg bw/d from the 2-yr rat study, using an assessment factor of 100 and corrected for oral absorption (35%). For the derivation of an ADI, the NOAEL of 21.7 mg/kg bw/d from the 2- yr rat study is applicable and including an assessment factor of 100, a value of 0.217 mg/kg bw/ day is derived.

The dietary exposure assessment concluded no concern based on the theoretical maximum consumption of 0.012 mg/kg bw/d using the food basket, which is 5.7% of the derived ADI. According to BPR guidance, no MRL needs to be derived if the dietary exposure is below 30% of the ADI.

Risk for industrial users

Not applicable. The product is not intended for use by industrial users.

Risk for professional users

Systemic effects Task/ Tier Systemic AEL Estimated Estimated Acceptabl Scenario NOAEL mg/kg uptake uptake/ e mg/kg bw/d mg/kg AEL (yes/no) bw/d bw/d (%) 1.Mixing&loa Tier 1 21.7 0.076 0.0120 15.76% yes ding mg/kg bw/day 1. Tier 2 21.7 0.076 0.0012 1.6% yes Mixing&loadi (glove mg/kg ng s, bw/day cover all) Scenario 2a Tier 1 21.7 0.076 0.550 mg/kg 724% no Professional bw/day user,

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Application by spraying (based on HEAdhoc 3) Scenario 2a Tier 2 21.7 0.076 0.055 mg/kg 72% yes Professional (glove bw/day user, s, Application cover by spraying all) (based on HEAdhoc 3) 3. Post- Tier 1 21.7 0.076 0.024 mg/kg 32% yes application – bw/day cleaning of equipment 3. Post- Tier 2 21.7 0.076 0.0024 3.2% yes application – (glove mg/kg cleaning of s, bw/day equipment cover all) 4. Workers Tier 1 21.7 0.076 0.022 mg/kg 29% yes accidentally bw/day touching treated manure

Biopren 50 LFL Fly larvicide concentrate has shown acceptable risks for all presented scenarios when considered separately, except for the application by spraying without protective equipment, calculated based on HEAdhoc recommendation 3. Professional users will always wear protective gloves and clothing. When PPE is worn, the risk is acceptable for all scenarios, based on all models.

Combined scenarios

As indicated above, it is realistic for professional users that the same person will be performing the mixing&loading, application and post-application tasks, therefore the assessment of combined risks from these scenarios is relevant. A combination of scenarios 1+2a+3 (mixing&loading, application and post-application) was therefore assessed.

Scenario 4 is not considered relevant for combined risks. Usually the pest control operator and the worker handling manure in livestock buildings is a different person and treatment of surfaces and handling of treated manure will not occur on the same day. A combination of these doses is therefore not relevant. Scenario 4 is considered separately in the risk assessment.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Scenarios Tier Systemic AEL Estimated Estimated Acceptable combined NOAEL mg/kg uptake uptake/ (yes/no) mg/kg bw/d mg/kg bw/d AEL bw/d (%) Scenarios Tier 1, 21.7 mg/kg 0.076 0.586 mg/kg 771% no 1+2a+3 no PPE bw/day mg/kg bw/day bw/day Scenarios Tier 2, 21.7 mg/kg 0.076 0.059 mg/kg 77% yes 1+2a+3 with bw/day mg/kg bw/day PPE bw/day

The risk of combined scenarios is acceptable when professional users wear personal protective equipment.

Local effects

S-methoprene does not produce local effects, neither after a single nor repeated exposure. BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE is classified with: H317: May cause an allergic skin reaction. Furthermore, the diluted product is classified with EUH208: Contains Polyethylene glycol (15)-hydroxystearate. May produce an allergic reaction. Based on this classification with H317 (during mixing and loading) and EUH208 (during spraying of the in-use dilution), P280: Wear protective gloves and protective clothing is prescribed. Taking this into account, risk due to this hazard is considered mitigated. Furthermore, the following general risk mitigation measures are included in the SPC: ‘Avoid contact with skin and eyes’ and ‘Wash hands thoroughly with hot water and soap after use’. No additional risk mitigation measures are considered needed.

Hazard Exposure Risk Hazard Effect Additiona P Who is Tasks, Potenti Frequen Potenti Relevant Conclusi catego s in l T exposed? uses, al cy and al RMM & on on ry terms relevant process exposu duration degree PPE risk of C&L hazard es re of of informati route potentia exposu on l re exposur e Mediu Skin - 1 Profession Mixing Skin 5 n.r. Gloves Accepta m Sens. 8 al, trained and minutes and ble 1, profession loading per day coveral H317 al prescrib ed

Addition al RMMs: - Avoid contact with skin and eyes.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

- Wash hands thoroug hly with hot water and soap after use.

Low EUH2 - 1 Profession Sprayin Skin 3 hours n.r. Gloves Accepta 08 8 al, trained g per day and ble profession coveral al prescrib ed

Addition al RMMs: - Avoid contact with skin and eyes. - Wash hands thoroug hly with hot water and soap after use.

Conclusion

On the basis of the scenarios presented above the risks to professional users are considered acceptable when personal protective equipment (i.e. gloves and coverall) is worn. Furthermore, the following risk mitigation measures are included in the SPC: ‘Avoid contact with skin and eyes’ and ‘Wash hands thoroughly with hot water and soap after use’.

Risk for non-professional users

Not applicable. BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE is not intended for non- professional use.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Risk for the general public

Systemic effects Task/ Tier Systemic AEL Estimated Estimated Acceptable Scenario NOAEL mg/kg uptake uptake/ (yes/no) mg/kg bw/d mg/kg AEL bw/d bw/d (%) Scenario [6], Tier 1 100 0.35 0.0257 7.3% yes school mg/kg children bw/day indirect exposure,

Children: A child may be exposed by dermal contact after accidentally touching the treated manure.

Calculating the risk for children as secondary exposure, the exposure was 7.3 % of the AEL. For the assessment of risks to children, the AELacute/medium term value was considered relevant.

From the above risk characterization it can be concluded that the BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE has a negligible risk on children entering the treated area.

Combined scenarios

Combined scenarios are not relevant for secondary exposure.

BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE contains only one active substance. However, besides the active substance a substance of concern was identified. As this substance of concern can only result in local effects, and S-methoprene only has systemic effects, no combined exposure of other effects are expected than already included in the risk assessment. Furthermore, it is not described that it should be used in combination with other formulations.

Local effects: S-methoprene does not produce local effects, neither after a single nor repeated exposure. The general public is only indirectly exposed to the diluted product. Athough BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE is classified with H317: May cause an allergic skin reaction, the diluted product would be classified with: EUH208: Contains Polyethylene glycol (15)-hydroxystearate. May produce an allergic reaction.. However, the diluted product is applied to manure. Furthermore, considering the EUH208 classification, this communicates that this effect could occur in already sensitized persons. A child may be exposed by dermal contact after accidentally touching the treated manure, and therefore, no local effects are expected for the general public.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Conclusion No adverse health effect is expected for the general public by use of BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE .

Risk for consumers via residues in food

During the use of the product no food, drinking water are exposed to the product. No risk is anticipated for consumers considering direct exposure, however a risk assessment is included for the indirect exposure of consumser via livestock exposure.

Risk characterisation from combined exposure to several active substances or substances of concern within a biocidal product

BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE contains only one active substance. However, besides the active substance a substance of concern was identified. As this substance of concern can only result in local effects, and s-methoprene only has systemic effects, no combined exposure other effects are expected than already included in the risk assessment. Furthermore, it is not described that it should be used in combination with other formulations

2.2.7 Risk assessment for animal health

An exposure assessment for livestock was presented for Biopren 50 LFL Fly Larvicide concentrate.

Based on the exposure levels calculated for relevant livestock (the product may only be used in piggeries, sheepfolds and cattle farms), as all exposure levels remained below the determined AEL, no unacceptable risks were identified for animal health.

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

2.2.8 Risk assessment for the environment

The environmental exposure of S-methoprene, formulated as capsule suspension (CS) is assessed. The product contains 3.75 % microencapsulated + 1.25 % ‘free’ S-methoprene. The insecticide function of the product is for the control of House fly ( Musca domestica ), Drone fly ( Eristalis tenax ) and Stable fly ( Stomoxys calcitrans ) through disruption of the development and metamorphosis of fly larvae and ultimately to prevent egg production. The trade name for the S-methoprene product is BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE. The preparation is proposed as a spray application for indoor (stables) and outdoor (manure pit, dunghill) use by professional users.

BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE is an S-methoprene based insect growth regulator, a juvenile hormone analogue. BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE prevents the emergence of adult flies, but has no effect on flies which have reached the pupal or adult stage prior to treatment.

The diluted preparation has to be sprayed onto the litter or the manure’s surface either with backpack sprayer, or with an appropriate device, eg. tractor mounted sprayers according to the following dosing: 200-240 mL/100 m 2.

For presenting the risk assessment of BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE on the environment all representative use scenarios (animals and housing) were selected for the exposure (and hazard) assessment.

The environmental exposure of S-methoprene was assessed in accordance with the Guidance on the Biocidal Product Regulation, Volume IV: Environment - Part B+C: Risk Assessment (Active Substances), European Chemicals Agency, Report no. ECHA-15-G-01- EN, Helsinki, Finland (2017), hereafter referred to as guidance, and the OECD ESD for Insecticides for Stables and Manure Storage Systems No.14 (2006). Concentrations for micro-organisms in a STP and the STP’s effluent were calculated with the model SimpleTreat version 3.0 by using the default values for parameters, unless otherwise noted. Release of active substance during the waste phase of the end-products is not assessed, because it is assumed that end-products to which the active substance is added are disposed as solid waste and usually incinerated. Possible pH effects on the environment were not considered, because the STP and receiving compartments are expected to have sufficient buffering.

In accordance with the Biocide Regulation, the life cycle of the product is incorporated into the risk assessment.

The risk characterisation is performed by comparing the predicted no effect concentration (PNEC), with the predicted environmental concentration (PEC). Considering the different ingredients in the product, only the active ingredient S-methoprene will cause risk for the environment and the risk characterization is therefore only performed for the active ingredient S-methoprene.

It has to be highlighted that there is no guide is available for testing the biodegradation in manure therefore all calculations is made without degradation of S-methoprene in manure as a worst case. However S-methoprene proved to degrade rapidly in the environment in the biodegradation study in soil (DT50 between 0.78 – 0.93 day in the deferent soil types) and in water sediment (total system: 0.87 and 1.32 day, water: 0.54 and 0.78 days, sediment: 6.72 and 3.74 days in pond and river system respectively). The relevant metabolites are also degrading fast (in the water sediment study metabolite M2 2.85 and 5.21 day; metabolite M3 1.21 and 1.91 day in the river and the pond system respectively).

NL Biopren 50 LFL FLY Larvicide concentrate PT 18

2.2.8.1 Effects assessment on the environment

PNEC determination on active ingredient S-methoprene: Details of PNEC determinations are provided in the S-methoprene Assessment Report (2013). PNEC’s relevant to risk characterisation in the affected compartments are as follows:

PNEC’s relevant to risk characterisation in the aquatic compartment (hydrosphere) are as follows: PNECSTP micro-organisms = 6.85 mg/L PNECaquatic (SW) = 0.00019 mg/L PNECsediment = 0.00038* mg/kg wwt PNECsoil = 0.148 mg/kg wwt

*At risk assessment stage, according to the guidance, the PNECsed is lowered by a factor of 10 due to the log Kow >5.

Information relating to the ecotoxicity of the biocidal product which is sufficient to enable a decision to be made concerning the classification of the product is required

According to the aquatic acute toxicity studies, the most sensitive species is Daphnia magna with an EC50 of 0.22 mg/L.

A summary of the aquatic ecotoxicological data presented for acute exposure indicates the following key endpoints:

Fish toxicity: 96 h LC50 in Zebrafish ( Brachydanio rerio ) = 4.26 mg/L Invertebrate toxicity: 48 h EC50 in Cladoceran ( Daphnia magna ) = 0.22 mg/L Algal toxicity: 72 h ErC50 in Selenastrum capricornutum = 2.264 mg/L

Based on the above acute toxicity results a chronic study was also commissioned on Daphnia magna .

Invertebrate toxicity: 21 d NOEC in Cladoceran ( Daphnia magna ) = 0.019 mg/L

The above ecotoxicity studies together with the biodegradation studies are sufficient for classification of S-methoprene.

Further Ecotoxicological studies

Further ecotoxicological studies on terrestrial organisms other than included in the current list of endpoints are submitted by the applicant for risk assessment purposes. The studies have been evaluated by the Dutch eCA. The evaluations are added to the IUCLID dossiers.New PNECs have been derived for soils. The PNECs for STP, surface water and sediment were taken from the current assessment report.

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Summary table - Further ecotoxicological studies Summary table of further ecotoxicological studies Method, Species End point Exposure Results Remark Refe- Guideline, (mg/kg dwt) s rence

GLP status, Design Dura NOEC EC 50 / Reliability -tion LC 50 OECD 222, Earthwor reproduction Reproduct 56 d 106 241 Nominal IUCLID ISO No.: m (56 d) ion test 5% OM section 11268-2GLP, (Eisenia 9.2.2.2 Reliability: 1 fetida ) mortality 28 d 213 404 (28 d) OECD 232, Collembol reproduction Reproduct 28 d 47 79.85 Nominal IUCLID ISO No.: a (56 d) ion test 5% OM section 11267-2GLP, (Folsomia 9.2.5 Reliability: 1 candida )

Conclusion used in Risk Assessment – Further ecotoxicological studies Value/conclusion From the terrestrial study results the Folsomia candida (Collembola) EC10 of 24.75 mg/kg dry soil was used for PNECsoil calculation. The study has been conducted with a high content of organic material in the artificial soil (i.e. 5% peat), the resulting endpoints have to be corrected for differences between the organic matter content of the test soil and that of the standard soil defined for biocides. For the latter, the standard average organic matter content of 3.4% is used to convert the endpoint to a standard soil for biocides resulting in a EC10 of 16.83 mg/kg soil dwt.

PNECsoil = 0.168 mg/kg dwt (= 0.148 mg/kg wwt) Justification for the The most sensitive organism among the species tested is the value/conclusion Collembolan. As two chronic studies were submitted, an assessment factor of 100 is applied because the available terrestrial ecotox data are derived for species belonging to a single trophic level

The value of PNECsoil in the Assessment Report of S-methoprene was estimated using the equilibrium partitioning method. (PNECsoil = 0.0003 mg/kg wwt). However new terrestrial studies with S-methoprene have been submitted at time of the BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE product dossier submission. The 28 days study result on Folsomia candida (Collembola) was a NOEC reproduction of 47 mg/kg soil dry weight and for Earthworm the NOEC reproduction was 106 mg/kg soil dry weight (56 day) and the NOEC mortality was 213 mg/kg soil dry weight (28 day). Since in the study with Folsomia candida already 20% inhibition of reproduction was observed at the level of the NOEC, the 28-d EC10 of 24.75 mg/kg dw is preferred over the NOEC of 47 mg/kg dw. The derivation of the EC10 is described in the study report and the dose-response curve seems to fit very wel to the observed effect percentages. The study has been conducted with a high content of organic material in the artificial soil (i.e. 5% peat). This means that the resulting endpoints have to be corrected for differences between the organic matter content of the test soil and that of the standard soil defined for biocides. For the latter, the standard average organic matter content of 3.4% is used to convert the endpoint to a standard soil for biocides resulting in a EC10 of 16.83 mg/kg soil

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dwt. As two chronic studies were submitted, an assessment factor of 100 is applied because the available terrestrial ecotox data are derived for species belonging to a single trophic level.

PNECsoil = 0.168 mg/kg dwt (= 0.148 mg/kg wwt)

Note from the CA evaluator: The derived PNEC is fully in line with guidance in Vol IV Part B+C (2017), but should be considered with care. From the PPP area it is well known that especially larvicides have very specific modes of action. Therefore possibly sensitive species have not been tested. At present, however, there are no tools on how to address this further.

Effects on any other specific, non-target organisms (flora and fauna) believed to be at risk (ADS)

Data waiving Information Not relevant requirement Justification No additional test on other target organisms is needed on the basis of intended uses, data available on the active substance or risk assessment.

Supervised trials to assess risks to non-target organisms under field conditions

Data waiving Information Not relevant requirement Justification Not applicable. The product is not applied directly on non-target organisms or soil.

Studies on acceptance by ingestion of the biocidal product by any non-target organisms thought to be at risk

Data waiving Information Not submitted requirement Justification No study on acceptance by ingestion of the biocidal product by any non-target organisms thought to be at risk is needed on the basis of intended uses, data available on the active substance or risk assessment.

Secondary ecological effect e.g. when a large proportion of a specific habitat type is treated (ADS)

Data waiving Information Not relevant requirement Justification No additional test on secondary ecological effect is needed on the basis of intended uses, data available on the active substance or risk assessment.

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Endocrine disruption activity of non-active substances

The Commission Delegated Regulation (EU) 2017/2100 specifying the scientific criteria for the determination of endocrine-disrupting properties (ED criteria) under Regulation (EU) No 528/2012 (BPR) establishes that the ED criteria become applicable by 7 June 2018 for biocides ( https://www.ctgb.nl/onderwerpen/hormoon-verstoorders ).

No further ecotoxicological studies are available for BIOPREN 4 GR FLY LARVICIDE GRANULE. The product was not tested for potential endocrine disruption properties. BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE contains the active substance s-methoprene and various co- formulants (see confidential annex).

For S-methoprene no ED assessment is required because for active substances which have been approved, the EU assessment should be followed. The Assessment Report (September 2016) states that S-methoprene is not included in the Commission staff working document on implementation of the EU Strategy for Endocrine Disrupters. Whilst S-methoprene is a juvenile (insect) hormone analogue, there is no evidence of any endocrine disruption potential in the human health or ecotoxicological studies presented in the dossier. As such it has been agreed that S-methoprene should be further assessed with regards to its potential endocrine disruptor properties once further guidance is available and preferably before the product authorisation stage. The conclusion of that assessment might lead to review of the active substance approval.

For the co-formulants a screening was performed by consulting: • ECHA data for identification of ED and PBT, under REACH or BPR or CLP • Identified as ED by United States EPA (https://comptox.epa.gov/dashboard/) • Identified as ED by the United Nations Environment (July 2017) Programme( http://wedocs.unep.org/bitstream/handle/20.500.11822/25634/edc_re port2.pdf?sequence=1&isAllowed=y and https://wedocs.unep.org/bitstream/handle/20.500.11822/25635/edc_report2_facts heet.pdf?sequence=1&isAllowed=y)

Only the co-formulant 2,6-di-tert-Butyl-p-cresol (CAS 128-37-0) triggered an alert for ED property. This co-formulant is included in the United Nations Environment Programme. And this is the same co-formulant that raised a concern based on the available toxicological information (see Section 2.2.6.1). As discussed in Section 2.2.6.1, CA NL considers that the ED assessment should await the outcome of the discussions at EU level. If this co-formulant is concluded to possess ED potency the authorisation granted for BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE needs to be re-evaluated.

Foreseeable routes of entry into the environment on the basis of the use envisaged

No significant direct release into the environment is expected from indoor professional use of BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE as a larvicide in animal housing. Emissions can occur from spreading treated manure on agricultural land, and from the potential release to municipal sewage treatment plants from cleaning of certain types of animal housing. Considering that most farms are not connected to the municipal sewer, waste water is often released to the manure depot instead. If present, residues may be released to individual sewage treatment plants. In all other cases release to the municipal sewer and subsequently to a sewage treatment plant (STP) is likely. During outdoor use, also surface water can be directly exposed by transport of insecticide residues via runoff and/or via agricultural drainage systems.

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Further studies on fate and behaviour in the environment (ADS)

Further studies on fate and behaviour have been submitted by the applicant for risk assessment puposes. It concerns a study on biodegradability in sewage sludge to determine if the active substance may be inherently biodegradable, one study on biodegradation in soils, and one study on degradation in water-sediment systems. These studies were already included as the post-approval studies in the CAR for S- methoprene (and discussed by member states during BPC-15 and BPC-16). The additional studies were submitted for commenting at BPC-15 (2016). eCA IE received comments from both DE and DK. After a three-week commenting period, it was addressed at BPC-16 that there were no further comments and IE updated the CAR addendum and LoEP based on the comments received. The evaluation of the study on biodegradation of S-methoprene in sewage sludge by the eCA IE is included below for completeness.

2.2.8.1.1 Biodegradability (ready/inherent)

Summary table - biodegradation studies (ready/inherent)

Metho Test Test Inoculum Additi Test Degradati Rema Referenc d, type 1 param onal sub- on rks e Guidel eter substr stanc Type Conc Ada Incu Deg ine, ate e en- p- ba- ree GLP conce tratio tati tion [%] status ntr. n on perio , d Realib ility

Modifie Inherent See Activa 100 No n/a 30 28 > See xxxx d MITI Biodegrad below ted ppm ne ppm days 70% below Inherent Test ability sludg Biodegrad (II), e ability of OECD S- guideli Methopren ne e In 302C, Modified GLP, MITI Test The (II). study xxxx , was unpublish consid ed report ered No.: accept 484.462.3 able 617 with a IUCLID Reliabil section ity 10.2 score of 1.

1 Test on inherent or ready biodegradability according to OECD criteria

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Conclusion used in Risk Assessment – Further studies on fate and behaviour in the environment Value/conclusion kdeg for the STP = 0/d Justification for the The biochemical oxygen demand (BOD) values were measured value/conclusion continuously during the experiment.

• Under the test conditions the percentage biodegradation of S- methoprene reached a mean of 4.2 % after 7 days, 24.5 % after 14 days, 77.5 % after 21 days and 85.8 % after 28 days based on its ThOD.

• The Inherent biodegradability of S-methoprene Technical was determined from the BOD measurement over 28 days according to OECD guideline 302C. The percentage of degradation of S-methoprene Technical was calculated using the BOD method, and supplemental chemical analysis was also carried out using HPLC method with UV detection on a Phenomenex, Luna 3µ C18 column.

From the BOD method the biodegradability was calculated to be 85.8% and by chemical analysis the degradation was found to be 74.2%. This modified MITI (II) test showed >70% degradation within 28 days. This represents inherent biodegradability (as specified in TGD). The failure to reach 70% within 14 days means that the specific inherent biodegradability criteria were not met and therefore that extrapolation of the results for use in STP models is not possible.

Leaching behaviour (ADS)

For product type 18 Insecticides a leaching study is not relevant.

Testing for distribution and dissipation in soil (ADS)

A soil dissipation study was submitted by the applicant for risk assessment purposes. A study considering aerobic degradation of S-methoprene in soil was submitted by the applicant for risk assessment puposes. This study is already included as post-approval study in the CAR for S-methoprene (and discussed by member states during BPC-15 and BPC-16). The additional studies were submitted for commenting at BPC-15 (2016). eCA IE received comments from both DE and DK. After a three-week commenting period, it was addressed at BPC-16 that there were no further comments and IE updated the CAR addendum and LoEP based on the comments received. The evaluation of the study on aerobic degradation of S-methoprene in soil by the eCA IE is included below for completeness.

Aerobic biodegradation Summary table – aerobic biodegradation in soil- laboratory study Test system

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Soil Soil pH OC Incu Method, Test Degr origin type % - Guideline, sub- a- Test batio Remar Referenc GLP stance datio type 1 n ks e status, concentr n perio Reliability a-tion DT * d 50

Aerobic Soil Germa Loam 5.5 1.6 0.52 120 1.76 See xxxx transformati degradati ny y 1 mg/kg days days below Degradati on in soil, on study sand on and OECD 307, Metabolis Germa Sand 6.0 0.6 1.38 GLP, The m in Four ny y 7 days study was Soils of Loam considered [14C] S- acceptable Germa Clay 7.1 1.7 1.50 methopre with a ny 3 days ne Reliability Incubated Germa Silt 6.2 1.1 1.57 score of 1. under ny Loam 9 3 days Aerobic Condition s. xxxx IUCLID section 10.2

* Geomean DT 50 for the four soils = 1.55 days

Conclusion used in Risk Assessment –Distribution and dissipation in soil Value/conclusion DT50 is 1.55 d at 12°C (geometric mean, n=4) Justification for the The route and rate of degradation of [ 14 C]S-methoprene in four soils value/conclusion incubated under aerobic conditions at 20 ± 2 °C in the dark were investigated.

14 • [ C]S-methoprene degraded rapidly in all four soils with DT 50 values of 0.93, 0.78, 0.79 and 0.83 days in soils I, II, III and IV, respectively. When coverted to 12 °C, this gives DT 50 values of 1.76, 1.38, 1.50 and 1.57 days in soils I, II, III and IV, respectively.

• Besides the parent compound one metabolite was identified by co-chromatography as Isopropyl (2E,4E)-11-hydroxy- 3,7,11-trimethyldodeca-2,4-dienoate (R4) reaching maximum mean amounts of 2.3%, 2.5%, 2.5% and 3.5% of applied on day 1 in soils I to IV, respectively.

• All other radioactive fractions were minor and transient, not exceeding 5.9% of the applied radioactivity at any sampling interval.

• High mineralisation of the radioactive residues to radiolabeled carbon dioxide was observed in all soils reaching maximum levels of 51.1%, 61.5%, 52.4% and 52.4% of the applied radioactivity for soils I to IV, respectively, on day 118.

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• The amount of non-extractable radioactivity was also significant, amounting to maximum mean values of 48.6% to 54.1% of the applied radioactivity during the 118-day incubation period. By the end of the study (day 118), the level of bound residues had declined. Organic matter fractionation on day 28 indicated that the majority of the non-extractable radioactivity was bound to the immobile humic acids and humins (26.7 – 41.4% of the applied radioactivity). Lower amounts of radioactivity (6.7 – 12.1% of applied) were detected in the more mobile fulvic acid fraction.

• The main degradation pathway of S-methoprene in soil proceeds via biodegradation beyond Isopropyl (2E,4E)-11- hydroxy-3,7,11-trimethyldodeca-2,4-dienoate (R4) as degradation inter-mediate and several minor and transient 14 fractions. In addition, a significant CO 2 production and formation of bound residues was observed.

Testing for distribution and dissipation in water and sediment (ADS)

A water/sediment dissipation study was submitted by the applicant for risk assessment puposes. This study is already included as post-approval study in the CAR for S- methoprene (and discussed by member states during BPC-15 and BPC-16). The additional studies were submitted for commenting at BPC-15 (2016). eCA IE received comments from both DE and DK. After a three-week commenting period, it was addressed at BPC-16 that there were no further comments and IE updated the CAR addendum and LoEP based on the comments received. The evaluation of the water/sediment dissipation study by the eCA IE is included below for completeness.

Water/sediment degradation test Summary table – fresh water/sediment degradation

Method, Test Test Test Incubat Degradation Remar Referen Guideline type 1 system substan ion ks ce (DT 50 /days) , GLP ce period Wat Sedim status, concent er ent Reliabilit ra-tion y

Aerobic Biodegrad River Rhine, 0.059 100 Parent See xxxx transform ation in Mumpf, AG, mg/kg days total 2.5 below Route ation in freshwater Switzerland total system 0 and Rate aquatic system Metabolite of sediment M2 Degrada systems, total 5.4 tion of OECD 308, system 0 [14C]S- GLP, The Metabolite Methopr study was M3 ene in considered total 2.2 Aerobic acceptable system 9 Aquatic with a Fröschweihe 0.061mg Parent Sedimen Reliability r pond, /kg total total 1.6 t score of 1. Möhlin, system system 5* Systems

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AG/Switzerla Metabolite . xxxx nd M2 unpublis total 9.8 hed system 8 report Metabolite no.: M3 D93728 total 3.6 IUCLID system 4 section 10.2

* parameter was derived using DFOP kinetics. Comparison of biphasic kinetic parameters with trigger cutoffs is not ideal. Therefore calculation of the DT 50 from the slow phase of the degradation yields a more conservative estimate of degradation:

° ° DT 50 = ln2/k 1 = 0.6931/0.1089 = 6.4 days @ T = 20 C or 12.1 days @ T = 12 C.

Conclusion used in Risk Assessment –distribution and dissipation in water and sediment

Value/conclusion DT 50 at 20°C are 0.65 d in water, 5.01 d in sediment, and 1.07 d for the whole system (geometric mean, n=2), corresponding to respectively 1.23, 9.50, and 2.03 d at 12°C. Justification for Data indicated that [ 14 C] S-Methoprene and metabolites degraded at a very rapid the rate and showed rapid dissipation from the total system. value/conclusion • Total recoveries of the applied radioactivity (material balances) averaged 92.7 ± 7.2% and 96.1 ± 10.7% of the applied radioactivity for the river and pond systems, respectively.

• After treatment, the majority of the radioactivity applied was detected in the water phase, representing 100.5% and 104.5% of the applied radioactivity for river and pond, respectively. In both systems the level of radioactivity in the water phase rapidly decreased over time, mainly by degradation and adsorption to the sediment. Within two days, the amount in the water phase had decreased to 38.2% and 28.5% of applied radioactivity for river and pond systems respectively. On Day 100, corresponding values were 1.2% and 0.6% of the applied radioactivity for river and pond, respectively.

• The total extractable radioactivity from sediments initially increased reaching maximum mean amounts of 21.2% (river) and 25.5% (pond) of applied after two days. Thereafter, levels decreased and on Day 100, 3.4% and 2.0% of the applied radioactivity was still extractable from the river and pond sediments, respectively. Soxhlet extractions were performed from Day 2 onwards, recovered up to 3.4% (river system, Day 2) and 4.7% (pond system, Day 14) of the applied radioactivity.

• The formation of radioactive carbon dioxide was significant, and constantly increased throughout incubation in both systems, reaching maximum mean amounts of 54.9% (river) and 67.5% (pond) of the applied radioactivity after 100 days of incubation. Organic volatile compounds never exceeded 0.6% of the applied radioactivity in either system.

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NL Biopren 50 LFL FLY Larvicide concentrate PT 18

• The relative amounts of [ 14 C] S-Methoprene and degradation products present in water and the sediment extracts were determined by chromatographic profiling by normal phase TLC.

• Significant metabolites in the water phase were M2 (R4 = NL 3.1.1.1) and M3 (R1 = FN 263). M2 reached up to 7.8% in the river and 6.2% in the pond waters by Day 2, and was detected until Day 21 in some samples. Maximum mean levels of M3 were 10.2% (river) and 5.8% pond, observed on Day 2. On all other sampling days, the concentrations of Metabolite M3 remained below 1%.

• The amounts of the test item reached maximum values after two days of incubation. At this interval the mean concentrations of [ 14 C] S-Methoprene in the river sediment were 16.6% and 20.8% in the pond sediment. These amounts decreased to 3.3% of applied in the river sediment and to 1.7% in the pond sediment by Day 49. In both sediments M2 (R4 = NL 3.1.1.1) and M3 (R1 = FN 263) were the predominant metabolites, but the levels did not exceed 1.9% of applied in either sediment, and had decreased to levels below 1% by Day 49. One additional metabolite M1 was detected which appeared to be less polar that the parent [ 14 C] S-Methoprene. M1 reached maximum mean levels 1.9% (river, Day 21) and 2.5% (pond, Day 14).

S-Methoprene degrades at a very rapid rate when applied to an aerobic aquatic environment. The primary route of degradation was mineralization (54.9- 67.5%). Significant formation of bound residues was observed (36.9- 41.0%). Acidic harsh extraction under reflux followed by organic matter fraction was conducted on the non-extracted residues from the Day 7 only released 1.6% or less of the total radioactivity. Overall, parent S-Methoprene as well as the two major metabolites M2 and M3 showed rapid dissipation from the total system. Neither S-Methoprene nor the metabolites appear to be persistent in the water- sediment system.

Test System @ T = 20°C Converted to T = S- 12°C - - 2 Methoprene Best M(0) K1(d K2(d g DT 50 DT 90 x DT DT 1 1 fit % ) ) (d)* (d)* error 50 90 (d)* (d)* River (water) SFO 95.9 0.8862 - - 0.78 2.60 0.554 1.48 4.93 Pond (water) SFO 99.1 1.2950 - - 0.54 1.78 0.982 1.02 3.38 River SFO 16.1 0.1855 - - 3.74 12.4 34.2 (sediment) 7.09 23.5 Pond SFO 20.9 0.1032 - - 6.72 22.3 7.23 (sediment) 12.8 42.3 River (total SFO 95.7 0.5235 - - 1.32 4.40 9.98 system) 2.50 8.34 Pond (total DFOP 99.1 0.109 1.223 0.2756 0.87 9.31 3.12 system) 1.65 17.7 M2 (NL 3.1.1.1) River (total SFO 0 0.2429 - - 2.85 9.48 18.5 system) 5.40 18.0 Pond (total SFO 0 0.1329 - - 5.21 17.32 28.2 system 9.88 32.9

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M3 (FN 263) River (total SFO 0 0.5745 - - 1.21 4.01 21.0 system) 2.29 7.60 Pond (total SFO 0 0.3613 - - 1.92 6.37 28.6 system 3.64 12.1 * The values quoted for water and sediiment phases refer to dissipation. The total system values refer to degradation

Testing for distribution and dissipation in air (ADS)

Data waiving Information Study on distribution and dissipation in air requirement Justification No accumulation of S-methoprene and its long range transport is expected. The possibility of long-range environmental transport (LRT) has been evaluated in the AR of S-methoprene and it was concluded that it was not expected that the substance will fulfil the screening criteria for the potential for LRT. S-methoprene is also not a persistent organic pollutant (POP).The physiochemical properties of S-methoprene do not suggest that this substance will pose a risk to the atmospheric environment. S-methoprene exhibits a medium to low volatility and sensitive to light. Thus an accumulation of S-methoprene in air and long range transport of the product is unlikely therefore no distribution and dissipation study was made.

If the biocidal product is to be sprayed near to surface waters then an overspray study may be required to assess risks to aquatic organisms or plants under field conditions (ADS)

BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE is used in animal stables and on manure only, therefore an overspray study is not required.

If the biocidal product is to be sprayed outside or if potential for large scale formation of dust is given then data on overspray behaviour may be required to assess risks to bees and non-target arthropods under field conditions (ADS)

BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE is used in animal stables and on manure only, therefore an overspray study is not required.

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2.2.8.2 Exposure assessment

General information Assessed PT PT 18 Scenario 1: Stable litter treatment Assessed scenarios Scenario 2: Manure pit treatment OECD Emission Scenario Document for Insecticides for ESD(s) used Stables and Manure Storage Systems. OECD report ENV/JM/MONO(2006)4. Scenario 1: Average consumption Approach Scenario 2: Average consumption Distribution in the Calculated based on Guidance on BPR Vol IV Part B+C environment (2017) and SimpleTreat version 3 Groundwater simulation Simulation for leaching with FOCUS PEARL 4.4.4 FOCUS SWASH 5.3 is used to calculate the PECs in surface Further higher tier water and sediment after STP sludge and manure application simulation to agricultural soil. Confidential Annexes No Scenario 1 and 2: Production: No Life cycle steps assessed Formulation No Use: Yes Service life: Yes PECs after ten years of successive manure application were calculated according to: - Recommendation of the BPC Ad hoc Working Group on Environmental Exposure. An Addendum to OECD SERIES ON EMISSION SCENARIO DOCUMENTS, Number 14: Emission Scenario Document for Insecticides for Stables and Manure Storage Systems ENV/JM/MONO(2006)4. (Agreed at the Environment Remarks session of WG-V-2015 on 27 November 2015). Item 7.7: Consultation of the BPC Working Group Environment. Proposal for revision of AHEE Recom WG V 2015 PT18. Calculation of PIECsoil_grassland after 10 year. (Agreed at the WG-I-2018, January 2018).Emission to the sewer was only considered for stable types and application methods for which a fraction to the STP is presented (TAB 2.0, ENV-168).

The environmental exposure of S-methoprene was assessed in accordance with the Guidance on BPR Vol IV Part B+C (2017) and the OECD ESD for Insecticides for Stables and Manure Storage Systems No.14. In accordance with the Biocides Regulation, the life cycle of the product is incorporated into the risk assessment. Degradation in manure was not taken into account. FOCUS PEARL 4.4.4. was used to estimate concentrations for S- methoprene in shallow groundwater. The physical-chemical parameters applied in the

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assessment for the different compartments (STP, water, sediment, soils and groundwater) are presented elsewhere.

The ESD distinguishes 18 types of farms (i1=1-18), which were in this risk assessment grouped in dairy cattle (i1=1), beef cattle (i1=2-3), pig farming (i1=4-6), and poultry (i1=7- 18). The final conclusion was based on the worst-case for each category as presented in the PAR, i.e. the stable type that results in the highest PECs. This is not necessarily the stable with the largest surface as the PECs are determined by nitrogen production as well. The worst-case stable types for each category are: • Dairy cows dairy cows (i1=1); • Beef cattle veal calves (i1=3); • Pig farming Sows in groups (i1=5); • Poultry Free range laying hens on litter (i1=11) for application in stables and turkeys (i1=16) for direct application on manure. Note that battery cages are not allowed in Europe (Regulation No 1999/74/EC) of the European Parliament) and therefore stable types i1=7-10 are not considered relevant. Nevertheless, none of these stable types are worst-case and therefore covered by the risk assessment for free range hens. The full risk assessment for all stable types is presented in the annex of the current PAR.

The diluted preparation has to be spread onto the manure’s surface either with back pack sprayer, or with an appropriate spraying device (only low pressure) at dose of 200-240 mL/100 m 2. Slatted floor: It is necessary to repeat treatment after each removal of dung. Deep litter: on every new layer of litter and after every 8 - 10 cm increase the treatment has to be repeated with the product.

Aquatic compartment (incl. sediment) and STP

Indoor use – STP route There is no direct exposure of surface water (including sediment) for the proposed use as an insecticide in stables when applied indoors. Indirect exposure of surface water and sediment is expected from indoor applications of the product in animal housings after wet cleaning where residues present on the stable’s floor or in equipment are removed and discharged to the sewer (certain poultry houses). In line with the TAB (version 2.0, ENV- 168) emission to the sewer is only considered for poultry for which a fraction to the sewer is presented in the ESD. There is no emission to the sewer from stables for dairy cattle, beef, and pigs.

The Simpletreat v.4.0 model is used to calculate distribution and degradation in the STP and to derive the PEC for the effluent, but in line with the TAB (ENV-9) the amount of suspended solids in the effluents is increased to 30 mg/L. The distribution and degree of removal of active substances in the STP is determined by the processes of biodegradation, adsorption onto sludge, removal due to sludge removal and volatilisation. Distribution and final PECs for water, sediment, and soils are subsequently calculated according to the guidance.

Indoor and outdoor use – manure/slurry route It may be expected that the majority of the active substance is released to the manure storage when manure and bedding materials are collected and/or stables are cleaned. Soils are therefore polluted when manure is applied as a fertiliser. The active substance may subsequently enter surface water in adjacent ditches when S-methoprene-enriched manure is applied as a soil fertiliser for agricultural land due to run-off or drainage. This route is assessed with FOCUS SWASH 5.5.3.

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The scenario in SWASH is similar to the PT18 groundwater assessment. As a worst-case for both arable land and grassland, the crop setting used is grassland. The application scheme is a distribution of the manure over 4 equal events. Application depth is 5 cm with soil incorporation and the application method is application to soil. The application dates are determined by an application scheme, which starts at March the 1st and ends September the 6th, with an interval of 53 days (and taking the 30 day SWASH requirement into account). All FOCUS scenarios are included. The application rate is set to the worst-case value of 1 kg/ha, covering spreading of manure on arable land and grassland, and sludge distribution on agricultural soils with a large margin of safety.

Soil compartment The PECs were calculated according to the ESD for insecticides for stables and manure storage systems. The exposure for application in stables was assessed according to the parameters for the application ‘larvicides in stables’ where the emission to the manure or slurry was based on the total surface of slatted areas, other areas, and manure areas inside (Table 5.3 of the ESD). In line with the TAB (ENV-168) the fractions to slurry and manure were summed for poultry. PECs resulting from application in manure were based on the default parameters as presented in Appendix 5 of the ESD. For application in slurry emission was only calculated for rectangular storage tanks being the realistic worst-case. Application on slurry stored in lagoons was not considered as such storage systems are unlikely in Europe to prevent distribution of pathogens, odour, and unacceptable emissions to the environment. Although the applicant has stated that the product is efficacious for twelve weeks, emission to the environment was still based on six applications during the fly season (ESD default) as repetitive treatment may be necessary. Degradation of the active compounds in manure or slurry during storage was not considered as no information is available. Degradation in soils between two successive manure or slurry applications was considered by applying the active substance’s half-life (DT 50 ) in soils.

According to the ESD for PT18, the deposition of active substances onto agricultural land (grassland) by manure/ slurry is estimated on the basis of emission standards for nitrogen or phosphor. Depending on the amount of nitrogen or phosphor in manure and the type of soil to which it is applied, these emission standards define the maximum amount of manure/slurry that can be applied per hectare and per year. The concentration in soil after manure/slurry application at maximum permissible rate (170 kg N/ha for both grassland and arable land and 110 kg P/ha for grassland and 85 kg P/ha for arable land) is calculated using the equations as proposed in the ESD for PT18. According to ENV 124 in the TAB (2017), it was decided to use the nitrogen immission standards from the EC Nitrates Directive (91/676/EC) of 170 kg N ha -1 yr -1 for all soils (arable land and grassland). Therefore, only the predicted environmental concentrations (PECs) based on the nitrogen emission standards are presented in the current PAR.

The PEC’s as calculated with the ESD represent the concentration after one manure application on arable land and one on grassland (Predicted Initial Environmental Concentrations, PIEC). However, agricultural soils are fertilised repeatedly and the active substance may consequently accumulate in soils after successive years of manure applications. Therefore, the concentrations presented in the current assessment report are the concentrations after ten years, i.e. ten manure applications on arable land and forty on grassland. Concentrations in soils after ten years were calculated according to the addendum for PT18 (insecticide in stables), although the no-manure time was increased from 206 to 365 days (agreed at WG-I-2018).

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The active substance disappears from soils between two subsequent manure events due to degradation, leaching, and evaporation. The leaching rate constants and resulting PECs were calculated according to the guidance based on the active substance’s physical-chemical parameters as presented elsewhere. The corresponding half-lives for leaching from the top soil layer are 7648 d in arable land (20 cm) and 3842 d in grassland (5 cm). Evaporation was not considered as the active substance does not easily evaporate from soil to air (low Henry’s law constant).

As the amount of disinfections per year and the surfaces to be disinfected strongly depend on the type of animal housed, emission to soils due to disinfection of stables varies among the different farm industries.

Groundwater compartment For refinement FOCUS PEARL (PEARL model © RIVM/Alterra, FOCUS v. 4.4.4) was used to estimate concentrations in shallow groundwater. The exposure scenario to soil did not include degradation of the active substance in manure. PEARL calculations do include aerobic degradation of active substance in soil.

The worst-case concentrations in soil were recalculated to kg/ha for manure spread on grassland and on arable land in order to be entered in PEARL. It was assumed that this concentration is equally distributed over 1 ha, in a soil layer with an incorporation depth of 0.2 m for arable land and of 0.05 m for grassland and a density of 1700 kg wwt/m 3. From these parameters, application rates expressed in kg /ha in manure in arable land or in grassland were calculated and used in PEARL. For arable land, a single land application to maize 20 days before crop emergence takes place. For grassland, four land applications to grassland/alfalfa at 1 st of March, 23 rd of April, 15 th of June and 7 th of August take place with an interval of 53 days take place each year. Anaerobic degradation in manure was not taken into account for the substance during manure storage. PEARL does include degradation in soil. The plant uptake factor was set at 0. The Q10 was set at 2.58. The activation energy was set at 65.4 kJ/mol. A simulation period of 20 years was chosen. The Freundlich exponent was set at 1.

Emission estimation

Two different emission pathways are described in the ESD for PT18 (2006): • Release via sewage treatment plant; • Release via slurry/manure

These pathways are all relevant for the environmental exposure assessment of BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE.

Scenario [1] Stable litter treatment

Input parameters for calculating the local emission Input Value Unit Remarks Scenario: Stable litter treatment Flies Type of insecticide - index i2: 3 (larvae) Application rate of biocidal product 2.4 mL/m² worst case (Qprod-uins)

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NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Concentration of active substance in 50 g/kg - the product (Fbioc) Type of application Spraying index i3: 1 Surface see remark m² The product is distributed over the stable’s floor. Emission is therefore based on the default total floor area as specified in the ESD

Scenario [2] Manure pit treatment

Input parameters for calculating the local emission Input Value Unit Remarks Scenario: Manure pit treatment Application rate of biocidal product 2.4 mL/m² worst case (Qprod-uins) Concentration of active substance in 50 g/kg - the product (Fbioc) Surface see remark m² Emission is based on the default areas for rectangular storage tanks

Identification of relevant receiving compartments based on the exposure pathway Fresh- Freshwate Sea- Seawater Ground- STP Air Soil Other water r sediment water sediment water Scenario 1: Stable yes yes yes yes yes no yes yes no litter treatment Scenario 2: Manure yes yes yes yes no no yes yes no pit treatment

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NL Biopren 50 LFL FLY Larvicide concentrate PT 18

Emission to the sewer The amount discarged to the sewer is presented below. The presented values are for turkeys (i1=16) being the worst-case.

Daily emission to the sewer Input Value Unit Remarks Stabble litter treatment 0.799E-02 kg/d - Application directly to Manure pit treatment - kg/d manure. Therefore no emission to the sewer.

The active substance’s properties applied for the exposure assessment are summarised below.

Input parameters for calculating the fate and distribution in the environment Input Value Unit Remarks Molecular weight of S-methoprene 310.48 g/mol - calculated with Melting point 53.8 °C Epiweb 4.1 Vapour pressure (20°C) 6.23E-04 Pa - Experimental solubility (20°C) 6.85 mg/L - Molar enthalpy of vaporisation 95 kj/mol - Molar enthalpy of dissolution 27 kj/mol - Henry’s Law Constant (20 °C) 0.0306 Pa/m 3/mol calculated Organic carbon-water partitioning - 8.76E+02 L/kg coefficient (K oc ) Organic organic matter-water 5.08E+02 L/kg Koc / 1.724 partitioning coefficient (K om ) Freundlich exponent 1.0 - Worst-case default Log Octanol/water partition 6.34 Log 10 calculated coefficient Not Ready Biodegradability biodegrada - - ble Half-life for biodegradation in soil 1.55 day - (Geometric mean, 12 °C)

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Whole system Half-life for biodegradation in water 2.5 day DT50 river (Geometric mean, 12 °C) system, AR (2016) Default worst-case Half-life for biodegradation in when whole 1000 day sediment (Geometric mean, 12 °C) system value is used for water. Half-life on crop canopy 10 d Default worst-case Plant uptake TSCF 0.0 - factor worst-case

Other SWASH parameters were copied from the EXSW0 substance in SPIN. Distribution in the sewage treatment plant was calculated according SimpleTreat. The values applied in the risk assessment are summarised below.

Calculated fate and distribution in the STP Compartment Percentage [%] Remarks Air 5.41E-02 Water 89.8 - Sludge 10.1 Degraded in STP n.r.

Calculations

Calculated PEC values

The PECs are presented below. All PECs are based after ten years of successive sewage sludge or manure application. Concentrations are based on the nitrogen emission standards for emission to the manure.

Summary table on calculated PEC values

PEC ST PEC wate PEC seawate PEC sed PEC seased PEC soil PEC GW PEC air P r r

3 [mg/L [mg/kg wwt [mg/kg wwt [mg/k [mg/m [mg/L] [mg/L] [μg/L] ] ] ] g wwt ] ]

via STP - Scenario 1, stable litter treatment (poultry)

3.59E- 3.58E- 1.46E- 7.11E-03 - - 0.01 -- 03 04 03

via slurry/manure to grassland - Scenario 1, stable litter treatment Dairy 2.38E- 3.71E- 4.72E-04 - - 2.38E-01 cattle -- 05 03 --

Beef 5.79E- 9.03E- 1.15E-03 - - 5.80E-01 cattle -- 05 03 --

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Pig 5.22E- 8.14E- farmin 1.03E-03 - - 5.23E-01 -- 05 03 -- g

5.77E- 8.99E- Poultry 1.14E-03 - - 5.77E-01 -- 05 03 --

via slurry/manure to arable land - Scenario 1, stable litter treatment Dairy 1.09E- 1.70E- 2.16E-04 - - 1.09E-01 cattle -- 05 03 --

Beef 2.65E- 4.14E- 5.26E-04 - - 2.66E-01 cattle -- 05 03 --

Pig 2.39E- 3.73E- farmin 4.75E-04 - - 2.40E-01 -- 05 03 -- g

2.64E- 4.12E- Poultry 5.24E-04 - - 2.65E-01 -- 05 03 --

via slurry/manure to grassland - Scenario 2, manure treatment Dairy 6.16E- 9.62E- -- 1.22E-03 - - 6.17E-01 -- cattle 05 03

Beef 2.31E- 3.60E- 2.31E+0 -- 4.58E-03 - - cattle 04 02 0 --

Pig 5.43E- 8.46E- farmin -- 1.08E-03 - - 5.43E-01 05 03 -- g

2.72E- 4.24E- Poultry -- 5.39E-04 - - 2.72E-01 05 03 --

via slurry/manure to arable land - Scenario 2, manure treatment Dairy 2.83E- 4.41E- -- 5.60E-04 - - 2.83E-01 cattle 05 03 --

Beef 1.06E- 1.65E- 1.06E+0 -- 2.10E-03 - - cattle 04 02 0 --

Pig 2.49E- 3.88E- farmin -- 4.93E-04 - - 2.49E-01 05 03 -- g

1.25E- 1.94E- Poultry -- 2.47E-04 - - 1.25E-01 05 03 --

A more realistic approach to calculate the sediment concentrations has been performed by using higher tier FOCUS SWASH modelling applying the input parameters as previously mentioned. PEC water and PEC sed following applications to grassland from all SWASH scenarios available for grass/alfafa are presented in the table below.

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Results for s-methoprene using FOCUS SWASH modelling for grassland

# Scenario PEC water PEC sed

[µg/L] [µg/kg dwt ] D1_ditch <0.001 <0.001 D1_stream <0.001 <0.001 D2_ditch <0.001 <0.001 D2_stream <0.001 <0.001 D3_ditch <0.001 <0.001 D4_pond <0.001 <0.001 D4_stream <0.001 <0.001 D5_pond <0.001 <0.001 D5_stream <0.001 <0.001 D6_ditch na na R1_pond na na R1_stream na na R2_stream <0.001 <0.001 R3_stream <0.001 <0.001

R4_stream na na na – scenario not available # In line with the agreed approach (WG ENV IV 2016) for the refinement of PECsed, the PECsw value should be converted to PECsed with the BPR guidance default distribution (see equation 53). This is due to differences in the suspended matter characteristics 8 between the BPR guidance and the FOCUS guidance. Therefore, the reported FOCUS TOXSWA (SWASH) PECsed values are not the correct values for biocidal product risk assessment, because the distribution between water and sediment was calculated differently. However, because FOCUS TOXSWA already included a distribution between water and sediment, thereby reducing the PECwater concentration, the direct conversion from the reported PECwater to PECsed would underestimate the actual PECsed value. Ideally, FOCUS TOXSWA should be recalculated with the BPR suspended matter characteristics. However, based on the reported risk values, it can be seen that a large margin of safety (see risk section) is available for both the water and sediment compartment. Therefore a recalculation of the distribution is not deemed necessary, because the outcome would remain the same.

The PEC values in groundwater presented in the above table were calculated in line with the approach of the Guidance on the BPR Volume IV Environment - Part B+C (2017) using the concentration in pore water in soil as indication for the groundwater level. Resulting PEC porewater concentrations exceed the 0.1 µg/L criterion for emission via manure or stable litter spread on grassland for scenario 2. Therefore it was concluded that a potential risk for groundwater exists and this needed to be assessed using higher tier methods, i.e. PEARL. The exposure scenario to soil did not include degradation of the active substance in manure. PEARL calculations do include aerobic degradation of active substance in soil.

The worst-case concentrations in soil for beef cattle in scenario 2 were recalculated to 3.06E- 02 kg/ha/year for manure spread on grassland and 5.61E-02 kg/ha/year for manure spread

8 Bulk density, volume, concentration

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NL Biopren 50 LFL FLY Larvicide concentrate PT 18 on arable land in order to be entered in PEARL. It was assumed that these concentrations are equally distributed over 1 ha, in a soil layer with an incorporation depth of 0.2 m for arable land and of 0.05 m for grassland and a density of 1700 kg wwt/m 3. For arable land, a single land application to maize 20 days before crop emergence takes place. For grassland, four land applications to grassland/alfalfa at 1 st of March, 23 rd of April, 15 th of June and 7 th of August take place with an interval of 53 days take place each year. Anaerobic degradation in manure was not taken into account for the substance during manure storage. PEARL does include degradation in soil. The plant uptake factor was set at 0. The Q10 was set at 2.58. The activation energy was set at 65.4 kJ/mol. A simulation period of 20 years was chosen. The Freundlich exponent was set at 1.

The resulting 80 th percentile of average annual groundwater concentrations at 1 m depth were < 0.001 µg/L at all nine PEARL locations.

Primary and secondary poisoning As the log Kow of the active substance is >3 (6.34 L/kg), the potential for bioaccumulation is considered high. As no experimental data is available, the expected bioconcentration factor (BCF) is calculated to be 516 L/kg. However, significant accumulation in terrestrial organisms (earthworms) is not expected as the active substance disappears quickly from soils (DT50 = 1.55 days at 12°C) and bioaccumulation of extreme hydrophobic compounds such as S-methoprene is slow (takes weeks to reach equilibrium), significant uptake is not expected.

The accompanied risks for birds and mammals is expected to be low regarding secondary poisoning as the active substance quickly disappears from soils as explained previously. It is furthermore not expected that birds and mammals are directly exposed to contaminated larvae as stables, manure storages, and slurry tanks are no common foraging areas. Hence the product meets the standards for the risk to birds and mammals. Primary and secondary poisoning is not expected for the intended uses.

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2.2.8.3 Risk characterisation

Atmosphere

Conclusion: Criteria for the examination of environmental risks to air are not specified in the form of a numerical standard. The assessment of potential impacts on air quality is aimed to minimize the risk for stratospheric ozone depletion. There are no indications that S-methoprene contributes to depletion of the ozone layer as the compound is not listed as ‘controlled substance’ in Annex I of Regulation (EC) No 1005/2009 of the European Parliament. Moreover, AOPwin calculates for the active substance a half life of 4.6 hours in air (OH timeframe 24 hrs/day, 0.5×10 6 OH radicals/cm 3). The calculated half life is below the trigger of 2 days, which is used as cut off value to identify chemicals that could be of potential concern for long range transport through the atmosphere. The environmental risk to air is therefore considered acceptable.

Sewage treatment plant (STP)

Summary table on calculated PEC/PNEC values

PEC/PNEC STP

Scenario 1, stable litter treatment (poultry)* < 0.001

* worst-case PECs for poultry are calculated for animal category 15 (parent broilers in rearing with grating floor) PECs calculated for poultry in battery cages were excluded.

Conclusion: The worst-case PEC/PNEC value for treatment of poultry stables is well below 1. From this PEC/PNEC value it can be concluded that no risk is identified for micro- organisms in the STP when poultry stables are treated with BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE.

Aquatic compartment

Emission of the active substance to surface water and sediment after application in animal housing occurs mainly via manure and waste water (STP), the latter only in some types of poultry houses.

Summary table on calculated PEC/PNEC values

PEC/PNEC water PEC/PNEC sediment Stable litter treatment via STP - (poultry) Scenario 1, stable litter treatment 1.89 18.7 (poultry)*

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NL Biopren 50 LFL FLY Larvicide concentrate PT 18

via slurry/manure to grassland Diary cattle 0.125 1.24

Beef cattle 0.305 3.02

Pig farming 0.275 2.72

Poultry 0.303 3.01 via slurry/manure to arable land Diary cattle 0.057 0.57

Beef cattle 0.14 1.39

Pig farming 0.126 1.25

Poultry 0.139 1.38 Manure treatment via slurry/manure to grassland Diary cattle 0.324 3.22

Beef cattle 1.22 12

Pig farming 0.286 2.83

Poultry 0.143 1.42 via slurry/manure to arable land Diary cattle 0.149 1.47

Beef cattle 0.557 5.52

Pig farming 0.131 1.3

Poultry 0.066 0.65

Conclusion: From this PEC/PNEC value it can be concluded that a risk is identified for water and sediment organisms when poultry stables are treated with BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE. The load to the STP for the worst-case animal category 15 (parent broilers in rearing with grating floor) is 0.00468 kg a.s./d, while for the best-case animal category 12 (broilers in free range with litter floor) the load to the STP is 0.00024 kg/d which is a factor 20 lower. Although the PEC/PNEC for sediment for this lower load to the STP is not exceeded (11/20 = 0.55) it is considered that there is acceptable risk for the STP for all poutlry houses in case the following risk mitigation measure is included in the proposed label: ” it is forbidden to use BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE in case effluent/wastewater of the stable and/or manure deposit can directly leach into the sewer.”

The first tier assessment for runoff from fertilised soils results in unacceptable risks for the aquatic environment. No risks for the sediment and surface water are identified considering higher tier modelling using FOCUS SWASH 5.3, see table below.

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Summary table on calculated PEC/PNEC values

PEC/PNEC water PEC/PNEC sediment Manure treatment via slurry/manure to grassland < 0.001 < 0.001 D1_ditch < 0.001 < 0.001 D1_stream < 0.001 < 0.001 D2_ditch < 0.001 < 0.001 D2_stream < 0.001 < 0.001 D3_ditch < 0.001 < 0.001 D4_pond < 0.001 < 0.001 D4_stream < 0.001 < 0.001 D5_pond < 0.001 < 0.001 D5_stream

D6_ditch na na

R1_pond na na

R1_stream na na < 0.001 < 0.001 R2_stream < 0.001 < 0.001 R3_stream

R4_stream na na na – scenario not available

Terrestrial compartment

Calculated PEC/PNEC values

** PEC /PNEC soil

via STP - Scenario 1, stable litter treatment (poultry) 0.010 via slurry/manure to grassland - Scenario 1, stable litter treatment

Dairy cattle 0.025

Beef cattle 0.061

Pig farming 0.055

Poultry 0.061 via slurry/manure to arable land - Scenario 1, stable litter treatment

Dairy cattle 0.012

Beef cattle 0.028

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Pig farming 0.025

Poultry 0.028 via slurry/manure to grassland - Scenario 2, manure treatment

Dairy cattle 0.065

Beef cattle 0.243

Pig farming 0.057

Poultry 0.029 via slurry/manure to arable land - Scenario 2, manure treatment

Dairy cattle 0.03

Beef cattle 0.112

Pig farming 0.026

Poultry 0.013

Conclusion: No risks are expected due to the distribution of sewage sludge or manure as all PECs are below the PNEC. Therefore, the standards for the terrestrial environment are met. No additional risk mitigations are required.

Non-target arthropods (including bees) The risk assessment to arthropods is considered to be similar to soil organisms due to their direct contact with soils. The standards for soil arthropods are therefore met. Because the active substance is expected to have a non systemic mode of action, secondary exposure of bees through pollen is considered negligible. Hence, the risk for bees is considered acceptable for the active substance.

Groundwater

Calculated PEC values

PEC ** (µg/L)

via STP - Scenario 1, stable litter treatment (poultry) 0.01 via slurry/manure to grassland - Scenario 1 and 2, stable litter or manure treatment Chateaudun < 0.001 Jokioinen < 0.001 Hamburg < 0.001 Kremsmuenster < 0.001 Okehampton < 0.001 Piacenza < 0.001 Porto < 0.001 Sevilla < 0.001

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Thiva < 0.001 via slurry/manure to arable land - Scenario 1 and 2, stable litter or manure treatment Chateaudun < 0.001 Jokioinen < 0.001 Hamburg < 0.001 Kremsmuenster < 0.001 Okehampton < 0.001 Piacenza < 0.001 Porto < 0.001 Sevilla < 0.001 Thiva < 0.001

Conclusion: After performing refined PEC groundwater calculations (using PEARL v. 4.4.4) for S- methoprene, it is concluded that the 0.1 µg/L criterion is not exceeded for STP sludge spread on land or manure or stable litter spread on grassland or arable land for all nine PEARL locations.

Primary and secondary poisoning

Conclusion: Primary poisoning is not expected for the intended uses. Although the BCF values suggest a high potential for bioaccumulation, accompanied risks for birds and mammals is expected to be low regarding secondary poisoning as the active substance quickly disappears from soils as explained previously. It is furthermore not expected that birds and mammals are directly exposed to granulates or contaminated larvae as stables, manure storages, and slurry tanks are no normal foraging areas. Hence the product meets the standards for the risk to birds and mammals.

Mixture toxicity

Screening step Screening Step 1: Identification of the concerned environmental compartments

The environmental compartments that are likely to be exposed are the STP, water, sediment, soil and the groundwater compartments.

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Screening Step 2: Identification of relevant substances

There is no relevant component in the mixture other than the active ingredient therefore the toxicity and the risk assessment of the active ingredient discussed in the previous paraghraphs will cover the toxicity to the environment and risk assessment of the mixture as well.

Screening Step 3: Screen on synergistic interactions

Screening step Significant exposure of environmental compartments? Y Number of relevant substances 1 Indication for synergistic effects for the product or its constituents in the literature? N

There is no other relevant component except the active ingredient, therefore no synergistic effect would occur. Futhermore the active ingredient S-methoprene is an Insect Growth Regulator (IGR) a Juvenile Hormone Analog (JHA). None of the components has the same mode of action and none of them are expected to have a synergistic effect.

Tiered approach No relevant component except the active ingredient that has an environmental toxicity was identified, therefore mixture risk assessment is covered by the risk assessment of the active ingredient S-methoprene.

Conclusion: The only relevant component in the mixture is the active ingredient. No risk was identified for all relevant environmental compartments when applying the product in animal housings.

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Aggregated exposure (combined for relevant emmission sources)

Decision tree on need for estimation of aggregated exposure

No aggregated Other yes Annual tonnage Biocide yes Biocidal no exposure regulatory of a.s. for use of a.s. < 10% specific emission estimation for areas biocide use of total? pattern a.s./b.p. required no no/unknown yes

Part 1 § Part 3 Other a.s. affected Uses of a.s./b.p. Part 2 Uses of a.s./b.p. within 1 PT Different user within >1 PTs Main constituent of a.s. is part of categories other a.s. Same a.s./b.p. in or different PTs

or Wide dispersive use a.s. is relevant metabolite or Overlap of other a.s ., in time and and vice versa Multiple b.p. space? for same purpose or no or yes

Different Different a.s. use/service life/waste form the same scenarios Aggregated No aggregated relevant metabolite exposure estimation exposure estimation required for a.s./b.p.* required for a.s./b.p. § Part 1 has to be checked for all PTs affected * a) aggregate only compartments and consider only PTs where overlap in time and space exists b) if production or formulation is within Europe, add a qualitative description of the respective environmental exposure e.g. in CAR

Figure 1: Decision tree on the need for estimation of aggregated exposure

Description: S-methoprene is also authorized according to the veterinary regulation. However, S- methoprene is mainly used in biocide products in the EU. These products are used in different areas, therefore there is no overlapping use of the products.

Decision steps:

Other regulatory areas?: Yes Biocide use of a.s. < 10 %? No Different user categories?: Yes Overlap in time and sapace?: No Conclusion: No aggregated exposure estimation required

Conclusion: No aggregated exposure estimation required based on the decision tree analysis.

Overall conclusion on the risk assessment for the environment of the product When used in accordance with the label instructions for use, BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE complies with the environmental standards and will not

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cause unacceptable effects to the environment provided that the following risk mitigation measure is included in the proposed label: ” it is forbidden to use BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE in case effluent/wastewater of the stable and/or manure deposit can directly leach into the sewer.”

2.2.9 Measures to protect man, animals and the environment

The product is used in animal housing on litter and manure to control house fly, drone fly and stable fly larvae. Detailed instructions are given on the product label.

Statement of risks arising and recommended methods and precautions concerning handling, storage, transport or fire: Methods and precautions concerning placing on the market Biopren 50 LFL fly larvicide concentrateis recommended for use by professional pest control operators. Specific training is not required for the use of Biopren 50 LFL fly larvicide concentrate. Methods and precautions concerning handling and use Eye protection: Not necessary

Respiratory protection: Not necessary

Other protective equipment: Not necessary

The usual precautionary measures for handling chemicals should be observed. Methods and precautions concerning storage Store at room temperature in areas that dry. Store away from heat, ignition sources and sunlight. Read and follow all precautions and instructions on the product label. Methods and precautions concerning transport Biopren 50 LFL fly larvicide concentrate is NOT classified as “Dangerous goods for transport”.

Proper shipping name: NON HAZARDOUS - not restricted Methods and precautions concerning fire

Extinguishing media: Water, carbon dioxide Fire fighting procedures: In case of fire no special measures are needed for fire fighting.

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Detailed procedures for the use and emergency measures in case of an accident: Specific treatment in case of an accident, e.g. first-aid measures, antidotes, medical treatment if available Eyes No known Flush with water for a few minutes adverse effects Skin No known Wash affected area with soap and adverse effects water.

Inhalatio Inhalation exposure not likely n Ingestion No known Rinse mouth, drink some water adverse effects

Emergency measures to protect the environment If spillage occurs sweep and transfer into waste containers for disposal. Clean the spill area with detergent and hot water. Contain the spill. Prevent from reaching surface waters or other water supplies. Avoid contact with skin and clothing.

Procedures for the destruction or decontamination of the biocidal product and its packaging: Procedures for waste management of the biocidal product and its packaging and where relevant, treated waste material for industry, professional users and the general public (non-professional users), e.g. possibility of reuse or recycling, neutralisation, conditions for controlled discharge, and incineration Disposal: The product and its container should be disposed of as hazardous waste. Prevent contamination of environment by wastes. Do not contaminate water, food or feed. Uncleaned packaging: Disposal in compliance with official regulations. RCA/CERCLA hazardous waste: Not listed.

No preliminary treatment of waste is necessary prior to disposal. Recycling is not an option.

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Possibility of neutralisation: Possibility of destruction or decontamination following release in the air Not applicable as the product is applied indoor (stables) and outdoor (manure pit) in controlled way. The active substance, S-methoprene is non-volatile (vapour pressure approximately 3.15 mPa) and therefore release to the air is not envisaged. Possibility of destruction or decontamination following release in water, including drinking water Release to water is not likely considering the use pattern of the product indoor (stables) and outdoor (manure pit) in controlled way. In the case of accidental exposure, prevent spillages from reaching surface waters or other water supplies. Contain the spill, sweep spillage and transfer into waste containers for disposal. The product contains 60.855 w/w%) water and the active ingredient S- Mmethoprene is rapidly degrading in water.

Possibility of destruction or decontamination following release in or on soil Direct application on soil of the product is not expected. Release to soil is via fertilizing the soil with manure considering the use pattern of the product: indoor (stables) and outdoor (manure pit) in controlled way. In the case of accidental exposure, prevent spillages from reaching surface waters or other water supplies. Contain the spill, sweep spillage and transfer into waste containers for disposal. The product contains 60.855 w/w%) water ingredients and the active ingredient S-methoprene is rapidly degrading in water.

Controlled incineration:

Not applicable

Measures to protect animals:

The product is used in animal housing on litter and manure therefore pets and wild animals are not expected to get in contact with the product. No risk mitigation is necessary.

Measures to protect the environment

When used in accordance with the label instructions for use, BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE complies with the environmental standards and will not cause unacceptable effects to the environment provided that the following risk mitigation measure is included in the proposed label: ” it is forbidden to use BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE in case effluent/wastewater of the stable and/or manure deposit can directly leach into the sewer.”

2.2.10 Assessment of a combination of biocidal products

BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE is not intended to be used with other products, therefore no combined assessment is necessary.

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2.2.11 Comparative assessment

Not relevant. S-methoprene is not a candidate for comparative assessment.

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3 ANNEXES 9

3.1 LIST OF STUDIES FOR THE BIOCIDAL PRODUCT

BIOPREN 50 LFL fly larvicide concentrate, xxxx , Unpublished Study No.: 484-170-0534 IUCLID xxxx 2015 Determination of the Y Bábolna Bio 3.4.1 Accelerated Storage Stability of Ltd

BIOPREN 50 LFL fly larvicide concentrate, xxxx , Unpublished Study No.: 484-160-0533 IUCLID xxxx 2015 Determination of the Y Bábolna Bio Ltd 3.5. Pourability of Biopren 50 LFL fly larvicide concentrate, xxxx Unpublished Study No.: 484-178-0607

9 When an annex in not relevant, please do not delete the title, but indicate the reason why the annex should not be included.

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Section No Author(s) Year Title Data Owner / Reference Source (where different Protection No from company) Claimed Company (Yes/No) Report No. GLP (where relevant) (Un)Published IUCLID xxxx 2015 Wet Sieve Test of Biopren 50 Y Bábolna Bio Ltd 3.5. LFL fly larvicide concentrate, xxxx Unpublished Study No.: 484-164-0606 IUCLID xxxx 2015 Determination of Flash Point of Y Bábolna Bio Ltd 3.5. Biopren 50 LFL fly larvicide concentrate, xxxx Unpublished Study No.: 484-150-0601 IUCLID xxxx 2015 Determination of the Surface Y Bábolna Bio Ltd 3.8. Tension of Biopren 50 LFL fly larvicide concentrate, xxxx , Unpublished, Study No.: 484-115-0605 IUCLID xxxx 2015 Determination of the Viscosity Y Bábolna Bio Ltd 3.9. of Biopren 50 LFL fly larvicide concentrate, xxxx , Unpublished, Study No.: 484-114-0604 IUCLID xxxx 2018 Determination of the pH Values Y Bábolna Bio Ltd. 3.2. of BIOPREN 50 LML mosquito larvicide concentrate, xxxx Unpublished Study no.: 484-122-4159, GLP IUCLID xxxx 2018 Determination of the pH Values Y Bábolna Bio Ltd. 3.2. of BIOPREN 50 LML mosquito larvicide concentrate (after storage) xxxx Unpublished Study no.: 484-122-4160, GLP xxxx 201 5 Determination of the Y Bábolna Bio Ltd. IUCLID Pourability of Biopren 50 LML 3.5. mosquito larvicide concentrate, xxxx Unpublished Study No.: 484-178-0546, GLP IUCLID xxxx 2017 Determination of the Y Bábolna Bio Ltd. 3.5. Pourability of BIOPREN 50 LML mosquito larvicide concentrate (after storage) xxxx Unpublished Study no.: 484-148-3436, GLP IUCLID xxxx 2018 Determination of Spontaneity Y Bábolna Bio Ltd. 3.5. of Dispersion of BIOPREN 50 LML mosquito larvicide concentrate, xxxx Unpublished Study No.: 484-168-3429, GLP IUCLID xxxx 2019 Determination of Spontaneity Y Bábolna Bio Ltd. 3.5. of Dispersion of BIOPREN 50 LML mosquito larvicide concentrate (after storage) xxxx Unpublished Study No.: 484-168-3430, GLP

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Section No Author(s) Year Title Data Owner / Reference Source (where different Protection No from company) Claimed Company (Yes/No) Report No. GLP (where relevant) (Un)Published IUCLID xxxx 2018 Determination of Suspensibility Y Bábolna Bio Ltd. 3.5. of BIOPREN 50 LML mosquito larvicide concentrate xxxx Unpublished Study No.: 484-169-3431,GLP IUCLID xxxx 2018 Determination of Suspensibility Y Bábolna Bio Ltd. 3.5. of BIOPREN 50 LML mosquito larvicide concentrate (after storage) xxxx Unpublished Study No.: 484-169-3432, GLP IUCLID xxxx 2015 Wet Sieve Test of BIOPREN 50 Y Bábolna Bio Ltd. 3.5. LML mosquito larvicide concentrate xxxx Unpublished Study no.: 484-164-0545, GLP IUCLID xxxx 2017 Wet Sieve Test of BIOPREN 50 Y Bábolna Bio Ltd. 3.5. LML mosquito larvicide concentrate (after storage) xxxx Unpublished Study no.: 484 -164 -3435, GLP IUCLID xxxx 2017 Determination of Persistent Y Bábolna Bio Ltd. 3.5. Foaming of BIOPREN 50 LML mosquito larvicide concentrate xxxx Unpublished Study No.: 484-158-3427 IUCLID xxxx 2017 Determination of Persistent Y Bábolna Bio Ltd. 3.5. Foaming of BIOPREN 50 LML mosquito larvicide concentrate (after storage) xxxx Unpublished Study No.: 484-158-3428 IUCLID xxxx 2018 Determination of the Auto - Y Bábolna Bio Ltd. 3.5 Ignition Temperature of BIOPREN 50 LML mosquito larvicide concentrate, xxxx Unpublished Study No.: 484- 153-3433, GLP IUCLID xxxx 2015 Validation of the Analytical Y Bábolna Bio Ltd 5. Method (HPLC) for the 484-100- determination of S-Methoprene 0535 in BIOPREN 50 LFL fly larvicide concentrate, xxxx Unpublished IUCLID xxxx 2007 S-methoprene Technical Y Bábolna Bio Ltd 5. Residue Analytical Method for CRA 119111 Determination in Tap Water, Surface Water and Ground , xxxx Unpublished IUCLID 5 xxxx 2012 Validation of the Analytical Y Bábolna Bio Ltd 484.102.361 Method for the Determination 8 of S-methoprene in Aquatic Formulations, xxxx Unpublished

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Section No Author(s) Year Title Data Owner / Reference Source (where different Protection No from company) Claimed Company (Yes/No) Report No. GLP (where relevant) (Un)Published IUCLID xxxx 2015 Validation of the Analytical Y Bábolna Bio Ltd 5 Method for the determination 484-100- of S-Methoprene in Edible Meat 0737 Matrices, xxxx Unpublished IUCLID xxxx 2014 FIELD TESTING OF AN Y Bábolna Bio Ltd 6.7. INSECTICIDE SPECIALITY INTENDED TO CONTROL FLIES IN BREEDING PREMISES, xxxx Study No: 1824/0914, Unpublished IUCLID xxxx 2015 FIELD TESTING OF AN Y Bábolna Bio Ltd 6.7. INSECTICIDE PRODUCT INTENDED TO CONTROL FLIES IN BREEDING PREMISES, xxxx Study No: 1931- FFT50LFL/0515, Unpublished IUCLID xxxx 2015 EFFICACY STUDY OF BIOPREN Y Bábolna Bio Ltd 6.7. 50 LFL FLY LARVICIDE CONCENTRATE IN DIFFERENT MANURES IN BIOLGICAL LABORATORY, xxxx Study No: 153.043.-044.-045. 153.048.- 049.-050. 153.053.-054.-055. 153.058.-059.-060., Unpublished IUCLID xxxx 2016 Laboratory measurement of Y Bábolna Bio Ltd 6.7. the effectiveness of an IGR- insecticide speciality intended for the control of th estable fly and eristalis tenax , xxxx 2069d/0416R, Unpublished IUCLID xxxx 2016 BIOLGICAL LABORATORY Y Bábolna Bio Ltd 6.7 EFFICACY STUDY OF BIOPREN 50 LFL FLY LARVICIDE CONCENTRATE IN DIFFERENT CONSISTENCY MANURES, 163.089-094 IUCLID xxxx 2015 Effects of S-methoprene Y Babolna Bio Ltd 9.2.2.2 technical on earthworm (Eisenia fetida ) reproduction in a chronic toxicity test, xxxx Study no. 484-222-0675, Unpublished IUCLID xxxx 2015 COLLEMBOLAN Y Bábolna Bio Ltd 9.2.5. REPRODUCTION TEST IN SOIL WITH S-METHOPRENE TECHNICAL, xxxx Study No: 484-232-0676, Unpublished

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Section No Author(s) Year Title Data Owner / Reference Source (where different Protection No from company) Claimed Company (Yes/No) Report No. GLP (where relevant) (Un)Published IUCLID xxxx 2011 Environmental distribution of Y Bábolna Bio Ltd 10.1. S-Methoprene (Level 1 Fugacity Calculator ver1.2 by Karl Nieman), xxxx , Study No: RIV2011/03/08, Unpublished IUCLID xxxx 2015 S-methoprene: Degradation Y Bábolna Bio Ltd 10.2. and Metabolism in Four Soils of [14C]S-methoprene Incubated under Aerobic Conditions, xxxx Study No: D93717, Unpublished IUCLID xxxx 2015 S-Methoprene: Route and Rate Y Bábolna Bio Ltd 10.2. of Degradation of [14C]S- Methoprene in Aerobic Aquatic Sediment Systems, xxxx Study No: D93728, Unpublished IUCLID 10.2. xxxx 2014 INHERENT BIODEGRADABILITY Y Babolna Bio Ltd OF S-METHOPRENE IN MODIFIED MITI TEST (II)

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3.2 OUTPUT TABLES FROM EXPOSURE ASSESSMENT TOOLS

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3.3 NEW INFORMATION ON THE ACTIVE SUBSTANCE New biodegradation studies:

1) S-methoprene: Degradation and Metabolism in Four Soils of [14C]S-methoprene Incubated under Aerobic Conditions, 2) S-Methoprene: Route and Rate of Degradation of [14C]S-Methoprene in Aerobic Aquatic Sediment Systems 3) INHERENT BIODEGRADABILITY OF S-METHOPRENE IN MODIFIED MITI TEST (II) See IUCLID section 10.2

New terrestrial non-target studies:

1) Collembolan reproduction test in soil with s-methoprene technical 2) Effects of S-methoprene technical on earthworm ( Eisenia fetida ) reproduction in a chronic toxicity test See IUCLID section 9.2

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3.4 RESIDUE BEHAVIOUR 3.5 SUMMARIES OF THE EFFICACY STUDIES (B.5.10.1-XX) 10 For summaries of the efficacy studies please refer to the efficacy section and IUCLID.

See IUCLID section 6.7

3.6 CONFIDENTIAL ANNEX Please see separate document.

3.7 ENVIRONMENTAL RISK ASSESSMENT

Manure.xlsx Stable Litter.xlsx

10 If an IUCLID file is not available, please indicate here the summaries of the efficacy studies.

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3.8 OTHER The Commission Delegated Regulation (EU) 2017/2100 specifying the scientific criteria for the determination of endocrine-disrupting properties (ED criteria) under Regulation (EU) No 528/2012 (BPR) establishes that the ED criteria become applicable by 7 June 2018 for biocides. No toxicological studies are available for the product Biopren 50 LFL FLY Larvicide cocnentrate. The product was not tested for potential endocrine disruption properties.

ED screening was performed by NL CA for all co-formulants, resulted in one potential alert. For the co-formulant 2,6-di-tert-Butyl-p-cresol (CAS 128-37-0) an ED concern has been raised by France https://echa.europa.eu/documents/10162/7ddda8e5-d66c-4fad-b502- 86cfdf2298bc and they propose to include this substance on the CoRAP list. CA NL cosider that the ED assessment for this co-formulant does not need to be included in the ED assessment of the PAR and await the outcome of the discussions at EU level.

NL CA concludes that Biopren 50 LFL FLY Larvicide concentrate does not possess co- formulants with ED alerts for human health.

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