Margosa Extract Product-type 18 (Insecticides, Acaricides and Products to control other Arthropods)

Transcription

1 Directive 98/8/EC concerning the placing biocidal products on the market Inclusion of active substances in Annex I or IA to Directive 98/8/EC Assessment Report Margosa Extract Product-type 18 (Insecticides, Acaricides and Products to control other Arthropods) 09/12/2011 Annex I - Germany

2 Margosa Extract (PT18) Assessment report Finalised in the Standing Committee on Biocidal Products at its meeting on 09/12/2011 in view of its inclusion in Annex I to Directive 98/8/EC CONTENTS 1. STATEMENT OF SUBJECT MATTER AND PURPOSE Procedure followed Purpose of the assessment report Overall conclusion in the context of Directive 98/8/EC OVERALL SUMMARY AND CONCLUSIONS Presentation of the Active Substance Identity, Physico-Chemical Properties & Methods of Analysis Intended Uses and Efficacy Classification and Labelling Summary of the Risk Assessment Human Health Risk Assessment Hazard identification Effects assessment Exposure assessment Risk characterisation Environmental Risk Assessment Fate and distribution in the environment Effects assessment PBT assessment Exposure assessment Risk characterisation List of endpoints DECISION Background to the Decision Decision regarding Inclusion in Annex I

3 3.3. Elements to be taken into account by Member States when authorising products Requirement for further information Updating this Assessment Report APPENDIX I: LIST OF ENDPOINTS APPENDIX II: LIST OF INTENDED USES APPENDIX III: LIST OF STUDIES

4 1. STATEMENT OF SUBJECT MATTER AND PURPOSE 1.1. Procedure followed This assessment report has been established as a result of the evaluation of margosa extract as product-type 18 (Insecticides, Acaricides and Products to control other Arthropods), carried out in the context of the work programme for the review of existing active substances provided for in Article 16(2) of Directive 98/8/EC concerning the placing of biocidal products on the market 1, with a view to the possible inclusion of this substance into Annex I or IA to the Directive. Margosa extract (CAS no ) was notified as an existing active substance, by Trifolio- M GmbH, hereafter referred to as the applicant, in product-type 18. Regulation (EC) No 1451/2007 of 4 December 2007, 2 which has repealed and replaced Commission Regulation (EC) No 2032/2003 of 4 November 2003, 3 lays down the detailed rules for the evaluation of dossiers and for the decision-making process in order to include or not an existing active substance into Annex I or IA to the Directive. In accordance with the provisions of Article 5(2) of Regulation (EC) No 2032/2003, Germany was designated as Rapporteur Member State to carry out the assessment on the basis of the dossier submitted by the applicant. The deadline for submission of a complete dossier for margosa extract as an active substance in Product Type 18 was , in accordance with Annex V of Regulation (EC) No 2032/2003. On , German competent authorities received a dossier from the applicant. The Rapporteur Member State accepted the dossier as complete for the purpose of the evaluation on On , the Rapporteur Member State submitted, in accordance with the provisions of Article 14(4) and (6) of Regulation (EC) No 1451/2007, to the Commission and the applicant a copy of the evaluation report, hereafter referred to as the competent authority report. The Commission made the report available to all Member States by electronic means on The competent authority report included a recommendation for the inclusion of margosa extract in Annex I to the Directive for PT18. 1 Directive 98/8/EC of the European Parliament and of the Council of 16 February 1998 concerning the placing biocidal products on the market. OJ L 123, , p.1 2 Commission Regulation (EC) No 1451/2007 of 4 December 2007 on the second phase of the 10-year work programme referred to in Article 16(2) of Directive 98/8/EC of the European Parliament and of the Council concerning the placing of biocidal products on the market. OJ L 325, , p. 3 3 Commission Regulation (EC) No 2032/2003 of 4 November 2003 on the second phase of the 10-year work programme referred to in Article 16(2) of Directive 98/8/EC of the European Parliament and of the Council concerning the placing of biocidal products on the market and amending Regulation (EC) No 1896/2000. OJ L 307, , p. 1 4

5 In accordance with Article 16 of Regulation (EC) No 1451/2007, the Commission made the competent authority report publicly available by electronic means on This report did not include such information that was to be treated as confidential in accordance with Article 19 of Directive 98/8/EC. In order to review the competent authority report and the comments received on it, consultations of technical experts from all Member States (peer review) were organised by the Commission. Revisions agreed upon were presented at technical and competent authority meetings and the competent authority report was amended accordingly. On the basis of the final competent authority report, the Commission proposed the inclusion of margosa extract in Annex I to Directive 98/8/EC and consulted the Standing Committee on Biocidal Product on 09/12/2011. In accordance with Article 15(4) of Regulation (EC) No 1451/2007, the present assessment report contains the conclusions of the Standing Committee on Biocidal Products, as finalised during its meeting held on 09/12/ Purpose of the assessment report This assessment report has been developed and finalised in support of the decision to include margosa extract in Annex I to Directive 98/8/EC for product-type 18. The aim of the assessment report is to facilitate the authorisation in Member States of individual biocidal products in product-type 18 that contain margosa extract. In their evaluation, Member States shall apply the provisions of Directive 98/8/EC, in particular the provisions of Article 5 as well as the common principles laid down in Annex VI. For the implementation of the common principles of Annex VI, the content and conclusions of this assessment report, which is available at the Commission website 4, shall be taken into account. However, where conclusions of this assessment report are based on data protected under the provisions of Directive 98/8/EC, such conclusions may not be used to the benefit of another applicant, unless access to these data has been granted Overall conclusion in the context of Directive 98/8/EC The overall conclusion from the evaluation is that it may be expected that there are products containing margosa extract for the product-type 18, which will fulfil the requirements laid down in Article 10(1) and (2) of Directive 98/8/EC. This conclusion is however subject to: i. compliance with the particular requirements in the following sections of this assessment report, 4 5

6 ii. the implementation of the provisions of Article 5(1) of Directive 98/8/EC, and iii. the common principles laid down in Annex VI to Directive 98/8/EC. Furthermore, these conclusions were reached within the framework of the uses that were proposed and supported by the applicant (see Appendix II). Extension of the use pattern beyond those described will require an evaluation at product authorisation level in order to establish whether the proposed extensions of use will satisfy the requirements of Article 5(1) and of the common principles laid down in Annex VI to Directive 98/8/EC. 6

7 2. OVERALL SUMMARY AND CONCLUSIONS 2.1. Presentation of the Active Substance Identity, Physico-Chemical Properties & Methods of Analysis The active substance margosa extract is an extract derived from ground seed kernels of the tropical neem tree Azadirachta indica using the manufacturing method developed by the applicant. Due to the complex nature of the botanical insecticide margosa extract, the main constituent - azadirachtin A (AzaA) - is used as the analytical lead compound. Margosa extract is a plant extract and consists mainly of the limonoids azadirachtin A, azadirachtin B, azadirachtin H, Desacetyl-Nimbin, Desacetyl-Salannin, Nimbin, Salannin together with co-extracted fatty acids and a small amount of water and other non-accounted materials. Margosa extract may contain traces of aflatoxins. Margosa extract is thermally stable up to 200 C, where it begins to decompose. The vapour pressure should be << 10-5 Pa based on the calculated vapour pressure of Pa for azadirachtin A. The water solubility and the partition coefficient can not be determined for extraction mixtures. Margosa extract is neither highly flammable (no relative self-ignition up to the melting point), explosive nor has oxidising properties. In conclusion, no hazard indication is required for the active substance margosa extract. Sufficiently validated analytical methods for the determination of azadirachtin A, the other limonids and impurities in the technical material are available. The determination of azadirachtin A is considered as the enforcement method, but methods for determination of all limonoids are available. Identity, Physico-chemical Properties and Method of Analysis of the Insecticide NeemAzal-T/S The biocidal insecticide NeemAzal-T/S is an emulsifiable concentrate containing further ingredients beside the active substance margosa extract. Due to the nature of the biocidal product it is not expected to exhibit any hazardous physical-chemical properties. It was noted that due to the limited thermal stability of the limonoids in margosa extract, the product may show limited shelf life, which can possibly be solved by recommending storage conditions (maximum temperature specified) and/or end use date. The analytical methods for NeemAzal-T/S in all relevant matrices are covered by the methods of azadirachtin A. Due to the complex nature of margosa extract, the choice of azadirachtin A as lead compound for residues in soil and water is accepted.

8 Intended Uses and Efficacy The assessment of the biocidal activity of the active substance demonstrates that it has a sufficient level of efficacy against the target organism and the evaluation of the summary data provided in support of the efficacy of the accompanying product, establishes that the product may be expected to be efficacious. In addition, in order to facilitate the work of Member States in granting or reviewing authorisations, and to apply adequately the provisions of Article 5(1) of Directive 98/8/EC and the common principles laid down in Annex VI of that Directive, the intended uses of the substance, as identified during the evaluation process, are listed in Appendix II. The aim of the treatment is to combat Thaumetopoea processionea caterpillars in order to prevent the formation of stinging and allergenic hair in the surrounding air which evoke allergic reaction in human and animal. As the dead caterpillars and the exuviae also contain the stinging hairs, margosa extract would not be able to directly reduce the number of hairs in the air, because it only kills the caterpillars. Direct reduction of the stinging hairs is only possible by physical methods. Nevertheless, larvae develop the stinging hairs in high amounts beginning with the 3 rd larval instar. Therefore, it is expected that the treatment will lead to a significant prevention of the formation of hairs in the air, when it is performed at a time when the predominant part of the caterpillars is in the 1 st or 2 nd larval instar. The efficacy is therefore determined by the killing effect of margosa extract, which in consequence leads to a prevention of the formation of stinging hairs in the air provided that the treatment is performed at an early larval stage (1 st or 2 nd larval instar). Therefore, the use of margosa extract has to be restricted to professionals, because special expertise is needed. The effectiveness of Neem extracts against different Order of insects is confirmed in the literature. The extract and the product deter feeding in insects and disturb the neurosecretory organs, inducing malformations and consequently death. The specific studies provided in the dossier are quite simple in methodology but confirm the efficacy in the laboratory and in the field of the product against different instar stages including 2 nd instar caterpillars of one specific target organism for which the product is claimed, i.e., caterpillars of T. processionea. Thus, in a laboratory study NeemAzal T/S is effective at a concentration of 0.3 % (corresponding to 0.009% margosa extract) which corresponds to the claimed application rate. 10L of the 0.3% diluted product are used per tree. Occurrence of Resistance Margosa extract is a plant extract from seed kernels of the Neem tree. Due to the complex composition of this plant extract and the complex mode of action which is not restricted to a single target site, the possibility of the development of resistance against this active ingredient is estimated as very low. 8

9 Classification and Labelling Classification and Labelling of Margosa extract Up to now there is no legal classification. Evaluation of the submitted data under Directive 98/8/EEC resulted in the following proposal for classification and labelling: Table 2-1 Proposed classification of margosa extract Classification Proposed Class of danger Xi Irritant Xn, Repr. Cat. 3 Harmful N Dangerous to the environment R phrases R 43 May cause sensitisation by skin contact R 63 Possible risk of harm to the unborn child R51 Toxic to aquatic organisms. R53 May cause long-term adverse effects in the aquatic environment. S phrases (S 2) Keep out of the reach of children S 36/37 Wear suitable protective clothing and gloves S60 This material and its container must be disposed of as hazardous waste. S61 Avoid release to the environment. Refer to special instructions / material safety data sheet. Remark: Classification according to the new Regulation (EC) 1272/2008 on classification, labelling and packaging of substances and mixtures (CLP): H317 (May cause an allergic skin reaction); H361d (Suspected of damaging fertility or the unborn child); H411 (Aquatic Chronic), category 2 In deviation to participant s proposal for classification and labelling, margosa extract is classified as sensitising by skin contact in view of the results in a Magnusson & Kligman Test. The observed malformations in a rat teratogenicity study are considered relevant for a classification as toxic to reproduction Category 3. The environmental classification is derived from the available acute tests with fish with the product NeemAzal-T/S. According to the findings of these tests, and taking into account also the results of the chronic fish test with margosa extract, there is evidence that the LC 50 to fish of margosa extract is in the range of > 1 mg a.s./l and < 10 mg a.s./l. margosa extract is degraded in aqueous systems by both biotic and abiotic processes, however, it does not fulfill the readily biodegradable -criteria. With regard to the environmental classification, the safety phrases S60, S61 are mandatory. The compound is neither highly flammable (no relative self-ignition up to the melting point), explosive nor has oxidising properties. In conclusion, no hazard indication is required for the physical and chemical properties for the active substance. 9

10 Classification and Labelling of NeemAzal-T/S Table 2-2 Classification according Directive 1999/45/EC Indication of danger R phrases Proposed classification of NeemAzal-T/S Proposed None Not required Not required S phrases None Not required The classification of NeemAzal-T/S is done on the basis of study results presented in the dossier, according to the principles of Directive 67/548/EEC and Directive 1999/45/EC. Besides the active substance margosa extract (content in the preparation: up to 4% w/w), the preparation contains formulation additives, which have not to be classified with regard to human health and environment. Margosa extract is classified as sensitising by skin contact (R 43) and Repr. Cat. 3 R 63. NeemAzal-T/S is not classified as sensitising based on the results of a maximisation test and it is not classified as R 63 because the concentration of margosa in NeemAzal is below the concentration limit of > 5 % in preparations. Although margosa extract is identified as dangerous to the environment (R51, R53), its concentration in the preparation is below the limit ( 25% w/w) that triggers the classification of the preparation. This evaluation is also supported by the available ecotoxicological data of the preparation showing an LC 50 value for fish of > 100 mg/l. Summary and conclusion: Classification and labelling of the insecticide NeemAzal T/S is not required Summary of the Risk Assessment Human Health Risk Assessment Hazard identification Absorption, Distribution, Excretion and Metabolism No studies on absorption, distribution, metabolism and excretion were submitted. Margosa extract is a plant extract and contains many known but also unknown constituents. The active compounds of the neem kernels are not known, the triterpenoids (known as limonoids) and among them the azadirachtins are considered as the most relevant in effectiveness against insects. Azadirachtin A is treated as the lead compound of margosa extract but it is unknown, 10

11 if this substance is also the most relevant with regard to toxicological aspects. Neither azadirachtin A nor any other limonoid is available as radioactive compound. Therefore, based on lack of technical feasibility, it is acceptable that no studies on metabolism and toxicokinetics were submitted. No guideline studies on dermal absorption were submitted. The submitted study (isolated perfused bovine udder model) fulfils the general criteria for scientific acceptance, and a low degree of dermal absorption was observed. However, as the extent of absorption for individual components of the extract was unknown, it was agreed at the TM-III-10 to use a value of 100 % dermal absorption for further risk assessment. Acute Toxicity Margosa extract is not acutely toxic when administered orally, dermally or by inhalation. It is neither irritating to the skin nor to the e of rabbits. Margosa extract is a skin sensitiser. Classification/labelling for sensitisation according to Directive 67/548/EEC: Xi; R43 Classification/labelling for sensitisation according to GHS, (EC) 1272/2008: H317 (May cause an allergic skin reaction) Short-term Toxicity In a 28-d rat feeding study with margosa extract, the animals of all dosed groups showed histological changes in the liver (eosinophilia, hepatocyte hypertrophy) increased liver weight and, additionally, thyroid follicular epithelial hypertrophy. Therefore, a NOAEL could not be identified, the lowest dose tested was 3200 ppm (300 mg/kg bw/d). In a 90-d rat feeding study with margosa extract, the top dose of 6400 ppm (490 mg/kg bw/d) induced hypertrophy of hepatocytes and thyroid follicular epithelial cells as well as an increase in liver and thyroid weight. Additionally, some changes in haematological parameters (prolonged APTT in males) were observed. At 1600 ppm (123 mg/kg bw/d), slight changes in a few clinical chemistry parameters and histological changes in liver histology (periportal fat deposition) in females were found. The NOAEL was 400 ppm (32 mg/kg bw/d). An appropriate repeated dose toxicity study with margosa extract in a second (non-rodent) species is lacking. The applicant justified non-submission by presenting published reports on the administration of neem seed products (water-washed neem seed kernel cake) to farm animals where no toxic effects were observed. Although the data obtained from these publications are not suitable to replace standard toxicity studies and uncertainties remain, it may be concluded that there was no indication for a high sensitivity or different target organs in non-rodents following (sub)chronic exposure. Therefore, non-submission was accepted. 11

12 Genotoxicity In vitro and in vivo tests provided no evidence for a genotoxic potential of margosa extract. Chronic Toxicity/ Carcinogenicity No carcinogenic potential and no adverse effects were found in the rat long-term toxicity study for the margosa extract, leading to a NOAEL of 6400 ppm (equivalent to 448 mg/kg bw/d). The mouse carcinogenicity study was conducted with the NeemAzal-F5 % formulation instead of margosa extract. This study presents several limitations and is therefore not fully acceptable. However, macroscopic and histopathological examinations did not reveal any substance-related increase in the occurrence of neoplastic lesions up to the highest dose tested (i.e. ca mg margosa extract/kg bw/d). Based on the absence of any carcinogenic potential up to the highest dose levels in the longterm studies in mice and rats, the absence of any other treatment-related adverse effects in the long-term rat study as well as in subchronic studies in a variety of other mammalian species, on the absence of a genotoxic potential in in vitro and in vivo genotoxicity tests, on the current discussion of the usefulness of the species mouse in long-term carcinogenicity studies and, finally, on animal welfare considerations no further long-term study in a second species is required. Reproduction Toxicity The developmental toxicity of margosa extract has been evaluated in the rat (Trifolio GmbH, here referred to as: NeemAzal technical) and in the rabbit performed with a different margosa extract. The results of the study evaluating the developmental toxicity in rabbit are read across to evaluate NeemAzal technical based on the similar toxicological profile of the extracts in subchronic rat studies. In the rat teratogenicity study with NeemAzal technical, an increased incidence of heart malformations in offspring were observed at 225 mg/kg bw/d and above. At 1000 mg/kg bw/d, maternal toxicity was evidenced by decreased body weight gain. The NOAEL for maternal toxicity was therefore 225 mg/kg bw/d. The NOAEL for teratogenicity and embryotoxicity was 50 mg/kg bw/d. In the rabbit teratogenicity study, the top dose of 500 mg/kg bw/d induced an increase in abortions and resorptions, and the remaining few live foetuses showed an increased incidence of malformations. At the same dose level, dams showed severe body weight loss from gestation days 6 to 12, resulting in a net body weight loss at study termination. At 100 mg/kg bw/d, body weight of dams was also slightly reduced between gestation days 6 and 12, but subsequently body weight gain increased, resulting in a net body weight gain. At 100 mg/kg bw/d, no effect on offspring was observed. The NOAEL for maternal toxicity was 20 mg/kg bw/d. The NOAEL for teratogenicity and embryotoxicity was 100 mg/kg bw/d. Malformations were observed in the high-dose group only and may be related to maternal toxicity. 12

13 Developmental toxicity was observed in both studies (rat and rabbit). In the rat study, specific malformations (affecting the heart) were found in the mid- and high-dose groups and an increased incidence of supernumerary rib was noted in the high-dose group. However, the heart malformations were also seen in the mid-dose group without clear adverse effects in the mothers; therefore they are not obviously produced by a non-substance-specific secondary mechanism. In a rat two-generation study with margosa extract, up to the highest dose tested of 750 ppm (51 mg/kg bw/d) treatment-related effects were observed neither in parental animals nor with respect to reproductive parameters. Therefore, the NOAEL was the highest dose tested of 750 ppm (51 mg/kg bw/d). In another supplementary (serious reporting deficiencies) rat two-generation study with NeemAzal-F 5 % (containing 5 % azadirachtin), increased relative testis and spleen weight was observed in all treated groups of the P 0.generation. Additionally, body weight gain in the mid- and high-dose groups was reduced. No treatment-related effects on reproductive performance were observed. Classification/labelling for reproduction toxicity according to Directive 67/548/EEC: Toxic to reproduction category 3, substances which cause concern for humans owing to possible developmental toxic effects, Xn R 63: Possible risk of harm to the unborn child. Classification/labelling for reproduction toxicity according to GHS, (EC) 1272/2008: H361d (Suspected of damaging fertility or the unborn child) Neurotoxicity In a non-glp study, three groups of six female chickens each were dosed daily by gavage with suspensions of margosa extract at dose levels of 0, 500, 750, and 1000 mg/kg bw/d for 21 days. The control group received distilled water. On day 22, 50 % of the birds were sacrificed and the remaining birds were observed for another 21 days. All birds were sacrificed on day 43. The following parameters of neurotoxicological relevance were investigated: a daily behavioural test for locomotive ataxia, activity of acetylcholine esterase in blood and serum on day 0, 23, and 43, histopathological examination of the brain (cerebrum, cerebellum, medulla oblongata), spinal cord (thoracic, cervical, lumbo-sacral) and sciatic nerve (proximal to distal length on either side) collected following sacrifice. In addition, animals were observed for clinical signs of toxicity, body weight and food consumption as well as number and weight of eggs laid. Blood was analysed for haematological and biochemical parameters. No treatment-induced effects were observed regarding neurotoxicity parameters or general toxicity. No treatment-related changes were noted up to the highest dose group and a NOEL of 1000 mg/kg bw/d was established in this study. 13

14 As margosa extract is neither an organophosphorous compound nor was the nervous system a target in the repeated dose toxicity studies, no further study is required. Mechanistic Studies No studies have been submitted. Waiving is justified considering that no specific effects are reported which would require a mechanistic clarification for further risk assessment. Medical Data Evaluation of the literature on neem demonstrates evidence of poisoning incidents and sideeffects in the use of neem products. "Margosa Oil" or "Neem Oil" is used as a traditional medicine in Asia and Africa. Case reports describe severe intoxications in children. Vomiting, drowsiness, convulsions, metabolic acidosis, and encephalopathy are among the reported signs of poisoning, autopsy of fatal cases revealed liver damage. According to some authors, the findings resemble those of Reye's syndrome. It is not possible to conclude on the toxic principles underlying the reported cases. According to some reports, aflatoxin residues may explain some poisoning incidents with the oil, while other authors assume an inherent toxicity of neem oil constituents. Anti-fertility (contraceptive and abortive) effects of oils and extracts are reported in studies with various mammalian species including humans (overview e.g. Schmutterer H., 2002, The Neem Tree, Mumbai). All clinical cases are reported for neem-derived materials, specifically for leaves and oil, but not for the margosa extract of the applicant (NeemAzal technical). Medical observations of workers in the production of margosa extract did not show adverse health effects in the three years observation period. Biocidal Product NeemAzal-T/S exhibited low acute oral, dermal and inhalative toxicity to male and female rats. The product NeemAzal-T/S is considered to be non- irritating to the skin and eye. Two studies for evaluation of skin sensitising potential (Magnusson-Kligman-Test, Buehler Test) lead to the conclusion that NeemAzal-T/S is considered as not sensitising to skin. No classification and labelling of the product NeemAzal-T/S is required. 14

15 Effects assessment Summarising the study results and all considerations above, margosa extract requires classification/labelling according to Directive 67/548/EEC as follows: Sensitisation by skin contact, Xi, R 43 Developmental toxicity Cat. 3, Xn, R 63 Based on malformations observed the teratogenicity study in rats, the NOAEL of 50 mg/kg bw/d (LOAEL = 225 mg/kg bw/d) is regarded as the relevant starting point for setting a systemic reference dose for acute exposure. By applying a default assessment factor of 100, a systemic acute Acceptable Exposure Level (AEL acute ) of 0.5 mg/kg bw/d is proposed for acute exposure to margosa extract. The NOAEL of 32 mg/kg bw/d from the 90-d toxicity studies in rats (LOAEL = 132 mg/kg bw/d) is regarded as the relevant starting point for setting a systemic reference dose for medium- and long-term exposure. By applying a default assessment factor of 100, a systemic medium-/long-term Acceptable Exposure Level (AEL medium-/long-term ) of 0.32 mg/kg bw/d is proposed for medium-/long-term exposure to margosa extract Exposure assessment Exposure of Professionals The active substance margosa extract is produced outside the EU. The biocidal product NeemAzal T/S (3 % a.s.) is produced within the EU. The occupational exposure is assessed for the formulation of the biocidal product. The results are not further evaluated for risk characterisation purposes under the requirements of the BPD. The following scenarios are covered by the exposure assessment in this report: Occupational exposure from use of biocidal product (spraying) 15

16 Secondary professional exposure The biocidal product is applied to control the oak processionary moth, Thaumetopoea processionea, first and second caterpillar stage. The predominant aim of control of this pest is to avoid contact to the hairs of oak processionary moth caterpillars during outdoor activities. Therefore, oak trees located in amenity areas and bearing these caterpillars have to be treated. The product will be applied with a hand-held spraying device from a lifting platform on crown level of the treated trees. Users are e.g. landscape conservation units, forest service, fire brigades. Before spraying the biocidal product NeemAzal T/S (3 % a.s.) is diluted to a spraying solution of % active substance. No information is supplied by the participant how many days an individual is exposed to the biocidal product. The rapporteur uses the following worst case assumption: For the personnel of landscape conservation units of communities application is restricted to 6 weeks or at most 36 days per year. In the case of specialized companies the period of application could be extended up to 8 weeks or up to 48 days per year. Two calculation models, the German BBA and the UK POEM were applied to estimate operator exposure. Due to the low vapour pressure of the active substance inhalation exposure during mixing & loading and post-application phase of the biocidal product was excluded from the exposure calculation. Moreover the formation of aerosol is not expected during these phases. The resulting level of exposure calculated by German BBA and UK POEM model are in the same order of magnitude. For risk assessment purposes the result of the German BBA model is taken forward. The resulting inhaled amount is mg a.s./person/day. Dermal exposure is expected during all phases of exposure. The assessment of the dermal exposure is based on the German BBA model for the mixing & loading and application phase. The dermal exposure during the post-application phase is assessed by the Riskofderm model. The resulting level of potential dermal exposure is 10.7 mg a.s./person/day. Exposure of bystanding professionals (secondary exposure) will be lower as compared to operator exposure. Since operator exposure was estimated to be well below the AEL even in the absence of PPE, no calculations or measurements are necessary to conclude that exposure of bystanding professionals will be below exposure limits. For further details please see Appendix I - List of endpoints - Acceptable exposure scenarios. Exposure of Non-Professionals Primary Non-Professional Exposure Primary exposure to non-professionals is not intended since the biocidal product in this dossier is foreseen for professional use only. However, it is likely that biocidal products for 16

17 non-professional use others than those against the caterpillars of the oak processionary moth will be submitted for national authorisation according to directive 98/8/EC. Therefore, a realistic worst-case-exposure assessment for non-professionals for the use in home and garden according to the German model was performed, resulting in a systemic exposure estimate of mg/kg bw/d. Medium-term is assumed since the non-professional user may apply the biocidal product frequently over the summer season. Thus, this is equivalent to 22.9% of the AEL medium-term (0.32 mg/kg bw/d). Thus, exposure of non-professionals using a biocidal product equivalent to NeemAzal-T/S would be acceptable in relation to human health. Secondary exposure to margosa-extract of the general public (consumer) Secondary exposure of the general public / bystanders is expected during professional application of the biocidal product. After application dermal exposure to the biocidal product is possible if persons re-enter and stay for a longer time period in areas, in which trees have been treated. Chronic secondary exposure is not expected for the general public. Table 2-3 Summary of secondary internal exposure of the general public (consumer) to margosa extract from NeemAzal-T/S Dermal exposure (mg/kg bw[/d]) Inhalation exposure (mg/kg bw[/d]) Modeled data Acute/Medium-term exposure internal dose Adults, bystanders Adults, re-entry, residents not relevant Infants, bystanders Infants, re-entry, residents not relevant Exposure via Residues in Food No residues in food are expected with the intended use. Combined Exposure Margosa extract is used at most a few times during a period of some weeks per year. Exposure of the general public can be neglected; therefore no concern is anticipated for a professional and non-professional combined exposure. However, margosa extract is used also as a pesticide in agriculture. It is not probable that the same persons are involved in the use of margosa extract both as a biocidal product and as a plant protection product in agriculture. However, this possibility cannot be excluded and therefore, as a worst case, it can be assumed 17

18 that the same persons would be involved in agricultural and biocidal use of margosa extract. The frequency of these combined exposures is not expected to go beyond medium-term timeframes; therefore a potentially combined agricultural and biocidal use of margosa extract is also adequately addressed in the risk characterisation of professional use in this report Risk characterisation Risk Assessment for Professionals In a first step (Tier 1) occupational risk assessment is based on the internal reference dose (AEL medium-/long-term ) of 0.32 mg/kg/day. A comparison with potential exposure gives a rough but cautious assessment to decide on concern (table 2-3). Under the specific conditions described there is no concern for the scenarios evaluated in this report. 18

19 Table 2-4 Risk characterisation for professionals using Tier 1 Exposure scenario Application of the biocidal product Mixing and loading Spray application Post-Application Specific conditions 45 ml NeemAzal T/S (3 % a.s.) is diluted with water in batches of 15 L - final concentration is % a.s. Form of exposure: liquid (3 % a.s.) Duration: 5-10 min Spraying of % a.s., 10 L per tree Form of exposure: aerosol (0.009 % a.s.) Duration: min. per task Frequency: Approximately trees may be treated per day, 8 weeks or at most 48 days per year Reloading of the spraying device, cleaning of spraying device Form of exposure: liquid (0.009% a.s.) Duration: min. potential exposure (external values) inhalation mg/kg bw/d dermal mg/kg bw/d Total potential internal dose (mg/kg bw/day) (1) Total internal body burden divided by AEL (2) Total (1) (2) 100% systemic availability after inhalation and dermal exposure AEL medium-/ long-term = 0.32 mg/kg/day Summary and conclusion: The following table summarise the risk characterisation outcome for professionals. 19

20 Table 2-5 Professional Users Primary Exposure (margosa) Exposure Scenario (indicate duration) estimated oral uptake [mg/kg b.w/day] Estimated Internal Exposure estimated inhalation uptake [mg/kg b.w/day] estimated dermal uptake [mg/kg b.w/day] estimated total uptake [mg/kg b.w/day] Relevant NOAEL/ LOAEL [mg/kg b.w/day] & Reference Value AF MOE re f MOE Exposure /AEL e.g: AEL (acute or medium or chronic) Tier 1 (no PPE) Mixing and loading 5-10 min./task max. 48 days /year Spray application min/task max. 48 days/year NOAEL= mg/kg/b.w./d AEL medium- /longterm=0.32 mg/kg b.w./d Postapplication min./task max. 48 days/year Total Tier 2 (Refinement, PPE or other risk mitigation measures Specify) Tier 2 is not required The occupational risk assessment for margosa extract is based upon the AEL approach and the estimate of potential occupational exposure. The risk assessment is considered to be sufficiently comprehensive and reliable for the purposes of Annex I inclusion of margosa extract. For the application of the biocidal product the risk assessment does not lead to concern. It is essential to recognize that this conclusion only applies to the active substance (margosa extract) in the biocidal product. From the point of view of occupational safety and health there is no risk-related reason for conditioning the requested Annex I inclusion for margosa extract. 20

21 However, against the background of experimental evidence of developmental toxicity, it needs to be stressed, that the AEL for margosa extract should be adhered to on every single day of an exposure period. Risk Assessment for Non-Professionals Primary exposure Since primary exposure to non-professionals is not expected a risk characterisation is not required. However, a realistic worst-case-exposure assessment for non-professionals for the use in home and garden according to the German model was performed, resulting in acceptable exposure estimates (table 2-5). Table 2-6 Non-Professional User Primary Exposure (margosa) Exposure Scenario (indicate duration) estimated oral uptake [mg/kg b.w/day] Estimated Internal Exposure estimated inhalation uptake [mg/kg b.w/day] estimated dermal uptake [mg/kg b.w/day] estimated total uptake [mg/kg b.w/day] Relevant NOAEL/ LOAEL [mg/kg b.w/day] & Reference Value AF MOE ref MOE Exposure /AEL e.g: AEL (acute or medium or chronic) Tier 1 (no PPE) Nonprofessional use, adult, Home & Garden, mixing & loading and application, repeated use, medium-term exposure x x x 10-2 NOAEL medium -term: 32 AEL mediumterm: Tier 2 (Refinement or other risk mitigation measures Specify) Tier 2 is not required 21

22 Secondary exposure For risk characterisation the systemic AEL medium-/long-term has been applied. This AEL derives from a 90-d rat study, which results in a NOAEL of 32 mg/kg/d. Applying an assessment factor of 100 leads to a systemic AEL medium-term of 0.32mg/kg bw/d. Although repeated exposure is unlikely the AEL medium-/long-term has been used for risk characterisation instead of the AEL acute representing worst case conditions. Additionally, the use of the AEL acute, which was derived from a developmental study, is not applicable for infants. In Tier 1 risk assessment the exposure estimates are below the AEL medium-term. Thus it is concluded that secondary exposure of non-professionals by the biocidal product NeemAzal- T/S is acceptable in relation to human health. Table 2-7 Summary Tier 1 risk assessment for secondary exposure Exposure Scenario (indicate duration) estimated oral uptake [mg/kg b.w./day] Estimated Internal Exposure estimated inhalation uptake [mg/kg b.w./day] estimated dermal uptake [mg/kg b.w./day] estimated total uptake [mg/kg b.w./day] Relevant NOAEL/ LOAEL [mg/kg b.w./day] & Reference Value AF MOE ref MOE Exposure /AEL e.g.: AEL (acute or medium- or long.-term) Bystander, adult, repeated exposure medium-term exposure x x x 10-1 NOAEL medium-term: 32 AEL mediumterm: Tier 1 (Worst Case) Medium-term Scenario 22

23 Bystander, infant, repeated exposure, medium-term exposure x x x 10-1 NOAEL medium-term: 32 AEL mediumterm: Resident, reentry, adult, repeated exposure medium-term exposure Resident, reentry, infant, repeated exposure, medium-term exposure x x 10-3 NOAEL medium-term: 32 AEL mediumterm: x x 10-2 NOAEL medium-term: 32 AEL mediumterm:

24 Table 2-8 Summary - risk assessment for secondary exposure Exposure scenario Exposure AEL medium-term Exposure MOE (mg/kg bw/d) (mg/kg bw/d) (% of AEL medium-term ) Acute/Medium-term exposure internal dose Adults, bystanders, dermal, inhalation Adults, re-entry, residents, dermal Adults, total Infants, bystanders, dermal, inhalation Infants, re-entry, residents, dermal Infants, total The exposure estimates are below the AEL medium-term. Thus it is concluded that secondary exposure of non-professionals by the biocidal product NeemAzal-T/S is acceptable in relation to human health Environmental Risk Assessment Fate and distribution in the environment The use of a non-labelled test substance and the non-feasibility of synthesising reference compounds result in an unavoidable limitation of the findings concerning the fate and behaviour of margosa extract i.e., the impossibility to determine the pathway of degradation of the test substance and the quality and quantity of its degradation products in a number of environmental fate studies (e.g., soil and water/sediment degradation, hydrolysis, aqueous photolysis). The findings for Aza A are considered in general to be representative at least for the other azadirachtins, i.e., Aza B and Aza H, of margosa extract, and therefore for a total portion of more than 40 % by weight of the active substance margosa extract. Biodegradation A total of five ready biodegradability tests were conducted: one with the formulated product NeemAzal-T/S, three with the active substance margosa extract (34 % azadirachtin A) and one with azadirachtin A. Only the product NeemAzal-T/S (containing 1 % azadirachtin A) 24

25 fulfilled the criteria of classification as readily biodegradable (i.e., > 60 % degradation within 10 days), probably attributable to the properties of the formulation additives, whilst neither margosa extract nor azadirachtin A proved to be readily biodegradable. No standard water/sediment study is available. Only literature data for the compound azadirachtin A were presented, which can only be regarded as additional information and will not be used for PEC estimation. Since radiolabelling of the main components of margosa extract was technically not feasible, the determination of bound residues, mineralisation rates and relevant metabolites was not possible. The extremely complex structure of azadirachtins also hampers the elucidation of metabolic and degradative pathways by exclusively analytical methods like HPLC, TLC, GC or spectroscopic methods. The dissipation rates of azadirachtin A in the different water systems are rapid to delayed. It can be assumed that hydrolysis is the main degradation process. The half-lives from literature data for azadirachtin A vary from 2.5 to 66 days converted to an average EU outdoor temperature of 12 C. Summarising the findings of three aerobic laboratory soil studies the DT 50 values for azadirachtin A varies between 1.9 and 37.9 days in six soils at 20 C resulting in a geometric mean of 5.45 days. Converted to an average EU outdoor temperature of 12 ºC the half-lives range between 3.6 and 71.9 days, the geometric mean is days (n=6). The DT 50 values of azadirachtin B amount to 39.7 and 99.0 days in two different soil types converted to 12 ºC. Since the tests were conducted with non-labelled material, neither mineralisation nor the formations of bound residues were quantifiable. In general quantification of the intermediate fractions and mass balance, however, were not possible with regard to the technical limitations linked to the work with unlabelled material. The study results conducted with unlabelled test substance show that azadirachtins and proposed metabolites indicate a rapid to delayed primary degradation. Abiotic Degradation The hydrolytic stability of azadirachtin A is strongly ph-dependent as indicated by a significant increase in the rate of degradation with increasing ph. The extrapolation of the test results to a temperature of 12 C using the Arrhenius equation yields a half-life of 117.7, 40.9, and 8.2 days at ph 4, 7, and 8, respectively. Hydrolysis products were not identified due to the technical limitations with regard to the impossibility of radiolabelling of the test substance and synthesis of reference substances. Aqueous photolytic half-lives for margosa extract were calculated based on the quantum yield and UV/VIS data from the direct phototransformation study in water of margosa extract and 25

26 parameters included in the computer model ABIWAS. The average half-life was estimated as 1.8 months for January and 5.5 days for July. With regard to the estimated half-life of h (equivalent to d) for azadirachtin A, long-term transport and accumulation in air are not to be expected. Furthermore, the tendency of azadirachtins, the major components of margosa extract, to enter the atmosphere is considered to be low taking into account both the vapour pressure of these compounds (3.6 x Pa) and the Henry s Law Constant (2.4 x Pa m 3 /mol). Distribution Results from two adsorption/desorption studies with azadirachtin A are available and give a median K oc value of 75.2 ml/g. Due to the low K oc value in the tested soils, azadirachtin A is slightly adsorbable to soil. Mobility The high mobility of azadirachtin A in soil as indicated by the low K OC is confirmed under the stringent conditions of the laboratory column leaching test, i.e., highly exaggerated concentration of substance applied to soil, maximum water saturation of soils at test start, watering with 200 mm rain within two days following test substance application. However, contamination of groundwater by azadirachtin A under actual use conditions seems to be unlikely taking into account its short degradation half-life in soil (see above). Bioaccumulation Bioconcentration factors for the aquatic (BCF fish = 2.5) and the terrestrial compartment (BCF earthworm = 1.1) were estimated on basis of the highest log P ow of 1.29 for azadirachtin B. The log P ow for azadirachtin A is 0.99 and the log P ow for azadirachtin H is analysed as The calculated BCF values indicate that a main component of margosa extract has a low potential to bioaccumulate in aquatic and terrestrial organisms and would pose no unacceptable risk of biomagnification in the food chain of either compartment Effects assessment Ecotoxicity tests both with the active substance margosa extract as well as the biocidal product NeemAzal-T/S are available. Tests with the product are considered as adequate for the effects assessment of the active substance as according to the available data on the formulation additives, the ecotoxicity of the b.p. is expected to be associated with the a.s. rather than any of those additives. 26

27 Aquatic Compartment The database of acceptable laboratory tests that is available for margosa extract and the product NeemAzal-T/S comprises both acute and long-term toxicity tests with fish, invertebrates and algae. The lowest effect value from the acute studies was obtained for fish (96h-LC50 = 4.14 mg a.s./l). However, the effect value for Daphnia was only a factor of 2 higher (48h-EC50 = 9.69 mg a.s./l). Algae are by orders less sensitive (72h-ErC50 = 1041 mg a.s./l; 72h-ErC10 = 332 mg a.s./l). In long-term tests performed with the product NeemAzal- T/S Chironomus riparius was most sensitive (28d-NOEC = mg a.s/l). For Daphina a 21d-NOEC of 0.1 mg a.s./l was derived while for fish the 28d-NOEC is 1.9 mg a.s./l. With the available data a PNEC water for margosa extract of mg a.s./l (0.6 µg a.s./l) was derived by applying an assessment factor of 10 to the lowest NOEC, as long-term tests with species from three trophic levels are available. Sediment PNEC sediment was derived from the PNEC water using the equilibrium partitioning method according to the TGD resulting in a PNEC sediment of mg a.s./kg ww (1.45 µg a.s./kg ww). Inhibition of microbial activity In a standard activated sludge respiration inhibition test with sludge from domestic sewage treatment plant a NOEC of 1000 mg a.s./l and an EC 50 of > 1000 mg a.s./l were determined. The risk to the micro-organism population of a sewage treatment plant can be characterised to be low to negligible regarding the results of these studies. From this data a PNEC micro-organisms of 100 mg a.s./l (nominal) was derived. Atmosphere For azadirachtins, i.e. the major components considered also responsible for the biological activity of margosa extract, physico-chemical properties have been determined resulting in low vapour pressure (3.6 x Pa) and the Henry s Law Constant (2.4 x Pa m 3 /mol). Thus, negligible volatilisation and transfer to air of azadirachtins are to be expected. Additionally, the chemical half-life of azadirachtin A in the troposphere was calculated to be 1.7 hours. According to these findings, accumulation and long-distance transport of margosa extract in the air followed by wet or dry deposition is not to be expected. Terrestrial Compartment All terrestrial tests were conducted with the product NeemAzal-T/S and the effect values are recalculated to the content of margosa extract. Acute tests with earthworms (14d-EC 50 = 12 mg a.s./kg dw) and plants (22d-EC 50 > 0.06 mg a.s./kg dw), a reproduction test with a 27