Endosulfan does not persist in living beings

The EFSA Journal (2005) 234, 1 - 31

Please note that this opinion, published on 7 April 2006, replaces the earlier version published 7 July 2005. The revision was considered necessary because the only study of oral toxicity of endosulfan in fish listed in the original opinion was found to be actually a study with endosulfan exposure of fish via water. Furthermore, two additional studies on fish exposed to endosulfan in feed became available to the Panel. The following items have been changed: • Section 5.2, 1st paragraph has been replaced by 3 new paragraphs,

• Conclusion, Adverse effects in animals, bullet on fish is replaced.

• Reference list: Naveed et al., 2004 has been replaced by Braunbeck and Appelbaum, 1999 and Coimbra et al., 2005.

The Panel on contaminants in the food chain is informed that there is an on-going oral toxicity study on Atlantic salmon exposed to endosulfan in the feed (Marc Berntssen, personal communication, The National Institute of Nutrition and Sea Food Research, Bergen, Norway, 2006). Based on the results of this study, the opinion might be updated again. This study is expected to be finalised in 2006.

http://www.efsa.eu.int of 31


OPINION OF THE SCIENTIFIC PANEL ON CONTAMINANTS IN THE FOOD CHAIN ON A REQUEST FROM THE COMMISSION RELATED TO ENDOSULFAN AS

UNDESIRABLE SUBSTANCE IN ANIMAL FEED

Question N° EFSA-Q-2003-066

Adopted on 20 June 2005

SUMMARY

Endosulfan is a non-systemic organochlorine pesticide that was developed and introduced in the mid 1950s. Endosulfan consists of α- and β-isomers that could be metabolised to endosulfan sulfate and endosulfan diol. In contrast to α- and β- endosulfan these metabolites are susceptible to photolysis. Endosulfan containing products still hold authorisation in seven member states of the European Community, but it is foreseen that the authorization of endosulfan will be withdrawn by the Member States before 1 February 2006. The use of endosulfan within the EU has steadily decreased during the recent years. Endosulfan is released into the environment mainly as a result of its use as a pesticide and is found in atmosphere, soil and sediment. Direct uptake from soil to plant as well as transport in plants is negligible.

In contrast to most related organochlorine pesticides, endosulfan has a less pronounced affinity to lipids. Consequently, biomagnification and accumulation of endosulfan, in terrestrial food chains, is less likely to occur.

Endosulfan is readily absorbed from the gastrointestinal tract and distributed to the kidneys and liver and to a lesser extent to other tissues. However, differences in distribution pattern between the isomers as well as metabolites have been reported.

Endosulfan residues are normally found in food and feed at low levels only. Detailed data on occurrence and temporal trends of endosulfan in feed are scarce.

Based on the limited data on animal exposure via feed produced according to good agricultural practice, it is not likely that terrestrial animals will be exposed to levels that could cause toxic effects.

Neurotoxic effects of endosulfan in both humans and animals are well documented. Exposure can induce a number of effects including liver and kidney toxicity, haematological effects, alterations in the immune system, and alterations in the reproductive organs.

Data from a limited number of samples suggest that intake of endosulfan by the general population, are far below the ADI of 6 µg/kg b.w. established by JMPR in 1998.

KEY WORDS: Endosulfan, α-endosulfan, β-endosulfan, endosulfan sulfate, analysis, toxicity, residues in feed and food, carry-over, ADI, environmental fate.



TABLE OF CONTENTS

LIST OF ABBREVIATIONS ............................................................................................................... 4 BACKGROUND ............................................................................................................................... 5 1. General Background............................................................................................................... 5 2. Specific Background .............................................................................................................. 6 TERMS OF REFERENCE ................................................................................................................... 8 ASSESSMENT ................................................................................................................................. 9 1. Introduction ............................................................................................................................ 9

1.1. Chemistry and use ......................................................................................................... 9 1.2. Environmental fate ...................................................................................................... 10 1.3. Toxicological findings in laboratory animals and hazard assessment for humans ..... 12

2. Methods of analysis.............................................................................................................. 14 3. Statutory limits ..................................................................................................................... 14 4. Occurrence in food and feed ................................................................................................ 14 5. Adverse effects on fish, livestock and pets, and exposure-response relationship................ 16

5.1. Introduction ................................................................................................................. 16 5.2. Fish .............................................................................................................................. 16 5.3. Ruminants.................................................................................................................... 17 5.4. Birds ............................................................................................................................ 18 5.5. Rabbits......................................................................................................................... 18 5.6. Dogs ............................................................................................................................ 18

6. Toxicokinetics and tissue disposition................................................................................... 19 6.1. Absorption................................................................................................................... 19 6.2. Distribution.................................................................................................................. 20 6.3. Metabolism.................................................................................................................. 20 6.4. Excretion ..................................................................................................................... 21

7. Carry over and tissue concentration ..................................................................................... 21 8. Human dietary exposure and comparison with ADI............................................................ 23 CONCLUSIONS ............................................................................................................................. 23 RECOMMENDATION..................................................................................................................... 25 REFERENCES ............................................................................................................................... 26 SCIENTIFIC PANEL MEMBERS....................................................................................................... 31 ACKNOWLEDGEMENT.................................................................................................................. 31 DOCUMENTATION PROVIDED TO EFSA ....................................................................................... 31



LIST OF ABBREVIATIONS

ADI Acceptable daily intake ATSDR Agency for Toxic Substances and Disease Registry B.w. Body weight CAS Chemical abstracts service ECD Electron capture detector FAO Food and Agricultural Organization FEFAC European Feed Manufacturers' Federation GABA Gamma-aminobutryic acid GC Gas chromatography IPCS International Programme on Chemical Safety JMPR Joint FAO/WHO meetings on pesticide residues LD50 Lethal dose that causes 50 % death of a group of test animals LOAEL Lowest observed adverse effect level Log Kow Logarithm of octanol-water partition coefficient MRL Maximum residue levels MS Mass spectrometry NOAEC No observed acute effect concentration NOAEL No observed adverse effect level PCB Polychlorinated Biphenyls POPs Persistent organic pollutants SCAN Scientific Committee on Animal Nutrition ULV Ultra-low volume WHO World Health Organization W.w. Wet weight



BACKGROUND

1. General Background

Directive 2002/32/EC of the European Parliament and of the Council of 7 May 2002 on undesirable substances in animal feed1 replaces since 1 August 2003 Council Directive 1999/29/EC of 22 April 1999 on the undesirable substances and products in animal nutrition2.

The main modifications can be summarised as follows

- extension of the scope of the Directive to include the possibility of establishing maximum limits for undesirable substances in feed additives.

- deletion of the existing possibility to dilute contaminated feed materials instead of decontamination or destruction (introduction of the principle of non-dilution).

- deletion of the possibility for derogation of the maximum limits for particular local reasons.

- introduction the possibility of the establishment of an action threshold triggering an investigation to identify the source of contamination (“early warning system”) and to take measures to reduce or eliminate the contamination (“pro-active approach”).

In particular the introduction of the principle of non-dilution is an important and far- reaching measure. In order to protect public and animal health, it is important that the overall contamination of the food and feed chain is reduced to a level as low as reasonably achievable providing a high level of public health and animal health protection. The deletion of the possibility of dilution is a powerful mean to stimulate all operators throughout the chain to apply the necessary prevention measures to avoid contamination as much as possible. The prohibition of dilution accompanied with the necessary control measures will effectively contribute to safer feed.

During the discussions in view of the adoption of Directive 2002/32/EC the Commission made the commitment to review the provisions laid down in Annex I on the basis of updated scientific risk assessments and taking into account the prohibition of any dilution of contaminated non-complying products intended for animal feed. The Commission has therefore requested the Scientific Committee on Animal Nutrition (SCAN) in March 2001 to provide these updated scientific risk assessments in order to enable the Commission to finalise this review as soon as possible (Question 121 on undesirable substances in feed)3.

It is worthwhile to note that Council Directive 1999/29/EC is a legal consolidation of Council Directive 74/63/EEC of 17 December 1973 on the undesirable substances in animal nutrition4,

1 OJ L140, 30.5.2002, p. 10 2 OJ L 115, 4.5.1999, p. 32 3 Summary record of the 135th SCAN Plenary meeting, Brussels, 21-22 March 2001, point 8 – New questions (

http://europa.eu.int/comm/food/fs/sc/scan/out61_en.pdf) 4 OJ L 38, 11.2.1974, p. 31



which has been frequently and substantially amended. Consequently, several of the provisions of the Annex to Directive 2002/32/EC date back from 1973.

The opinion on undesirable substances in feed, adopted by SCAN on 20 February 2003 and updated on 25 April 20035 provides a comprehensive overview on the possible risks for animal and public health as the consequence of the presence of undesirable substances in animal feed.

It was nevertheless acknowledged by SCAN itself for several undesirable substances and by the Standing Committee on the Food Chain and Animal Health that additional detailed risk assessments are necessary to enable a complete review of the provisions in the Annex, including the establishment of maximum levels for undesirable substances currently not listed.

2. Specific Background

Endosulfan is an organochlorine insecticide. Contrary to some other organochlorine pesticides, endosulfan does not accumulate in the food chain and is eliminated rapidly from the body. Endosulfan is highly toxic for some aquatic species, particular fish.

Contrary to the other pesticides listed in the Annex to Directive 2002/32/EC, endosulfan is still in use as a pesticide.

Current EU legislation on maximum residue levels (MRLs) for pesticides is derived from/based on four Council Directives

- Council Directive 76/895/EEC of 23 November 1976 relating to the fixing of maximum levels for pesticide residues in and on fruit and vegetables6

- Council Directive 86/362/EEC of 24 July 1986 on the fixing of maximum residue levels for pesticide residues in and on cereals7

- Council Directive 86/363/EEC of 24 July 1986 on the fixing of maximum residue levels for pesticide residues in and on foodstuffs of animal origin8

- Council Directive 90/642/EEC of 27 November 1990 on the fixing of maximum residue levels for pesticide residues in and on certain products of plant origin, including fruits and vegetables9.

- Regulation (EC) No 396/2005 of the European Parliament and of the Council of 23 February 2005 on maximum residue levels of pesticides in or on food and feed of

5 Opinion of the Scientific Committee on Animal Nutrition on Undesirable Substances in Feed, adopted on 20

February 2003, updated on 25 April 2003 (http://europa.eu.int/comm/food/fs/sc/scan/out126_bis_en.pdf) 6 OJ L 340, 9.12.1976, p.26 7 OJ L 221, 7.8.1986, p. 37 8 OJ L 221, 7.8.1986, p. 43 9 OJ L 350, 14.12.1990, p. 71



plant and animal origin and amending Council Directive 91/414/EEC which will repeal the four Council Directives10.

Until 1997, MRLs were fixed only for raw commodities. Council Directive 1997/41/EC of 25 June 199711 amending the above mentioned Directives, provided for a system applicable from 1 January 1999 to set MRLs in processed products and composite foodstuffs, based on the MRLs fixed for the raw agricultural products. MRLs for processed products and composite foodstuffs are calculated on the basis of the MRL set for the agricultural commodity by application of an appropriate dilution or concentration factor and for composite foodstuffs MRLs are calculated taking into account the relative concentrations of the ingredients in the composite foodstuffs.

As the consequence of the coming into force of Directive 1997/41/EC, the pesticide residue legislation applies also to animal feeding stuffs since 1 January 1999. However some problems have been observed in implementing the pesticide residue legislation. The following problems have already been identified:

- compound feed is composed of a relatively high number of ingredients of which several are processed products (by-products). It is not obvious to know what MRL is applicable to such compound feed as it involves many calculations and uncertainties and “unknowns” (processing factors).

- pesticide residue legislation does not cover products of marine origin which are regularly used in animal feed (no direct application).

- pesticide residue legislation does not cover products typically for animal feed (no food use) such as pastures, roughages, forages, fish oil and fish meal.

Therefore it is appropriate to include in the list of undesirable substances maximum levels for some pesticides, in particular those of relevance for animal health or for public health through carry over from feed to food of animal origin

As already mentioned, endosulfan is listed in the Annex to Directive 2002/32/EC. For comparison the current provisions in the EU-pesticide residue legislation are mentioned.

10 OJ L 70, 16/03/2005, p. 1 11 OJ L 184, 12/07/1997, p. 33



Directive 2002/32/EC EU-Pesticide residue legislation ML relative to a feeding stuff with a moisture content of 12 %

MRL applicable to the product as marketed

Product mg/kg Product mg/kg Maize 0.2 Cotton seeds 0.3 Oilseeds 0.5 Soybean seeds 0.5 Complete feeding stuffs for fish 0.005 Other oilseeds 0.1* All other feeding stuffs 0.1 Potatoes 0.05* Tea 30 Hops 0.1* Cereals 0.05* Citrus, berries 0.5 Pome fruit 0.3 Tree nuts 0.1* Peppers 1 Tomatoes 0.5 Other vegetables 0.05* Meat (fat) 0.1 Milk 0.004 Eggs 0.1* *lower limit of analytical determination

In the current provisions in Directive 2002/32/EC there are apparently some inconsistencies. Whereas for example a maximum level for endosulfan is fixed at 0.5 mg/kg for soybean seeds (oilseeds) the resulting soybean oil (in which endosulfan concentrates) has to comply with 0.1 mg/kg (all other feeding stuffs) in case it is used for animal feed. For foodstuffs a processing factor can be applied.

It is important that these provisions concerning endosulfan are completely reviewed in the framework of Directive 2002/32/EC. A risk assessment on the presence of endosulfan in animal feed, in particular fish feed and fish feed ingredients, should be undertaken as a priority.

TERMS OF REFERENCE

The European Commission requests the EFSA to provide a scientific opinion on the presence of endosulfan in animal feed.

This scientific opinion should comprise the

- determination of the toxic exposure levels (daily exposure) of endosulfan for the different animal species of relevance (difference in sensitivity between animal species, with particular attention to farmed fish species) above which

- signs of toxicity can be observed (animal health/impact on animal health)



- the level of transfer/carry over of endosulfan from the feed to the products of animal origin results in unacceptable levels of endosulfan or of its metabolites in the products of animal origin in view of providing a high level of public health protection.

- identification of feed materials which could be considered as sources of contamination by endosulfan and the characterisation, insofar as possible, of the distribution of levels of contamination

- assessment of the contribution of the different identified feed materials as sources of contamination by endosulfan

- to the overall exposure of the different relevant animal species (with particular attention to farmed fish species) to endosulfan,

- to the impact on animal health,

- to the contamination of food of animal origin (the impact on public health), taking into account dietary variations and carry over rates.

- identification of eventual gaps in the available data which need to be filled in order to complete the evaluation.

ASSESSMENT

1. Introduction

1.1. Chemistry and use

Endosulfan is an organochlorine pesticide that was developed and introduced in the mid 1950s. World wide production of endosulfan in the middle of the 1980’s was estimated at 10,000 tons/year (ATSDR, 2000). Within the European Union there is currently only one producer located in Frankfurt (Germany) which produces endosulfan. The manufactured volume of endosulfan at this site currently amounts for approximately 4,000 tons/year and a major part is exported for use in tropical and subtropical regions such as Latin America, Caribbean and Southeast Asia. Endosulfan has also been reported to be produced in Israel, India, South Korea, and China (Umweltbundesamt, 2004).

CAS numbers are for technical endosulfan 115-29-7, α-endosulfan 959-98-8, β-endosulfan 33213-65-9, and endosulfan sulfate 1031-07-8.

Technical endosulfan is obtained through the Diels-Alder addition of hexachlorocylopentadiene and cis-butene-1,4-diol, followed by reaction of the addition-product with thionyl chloride. It mainly consists of a mixture of two stereoisomers named α- and β- endosulfan in the approximate ratio of 70:30 (Figure 1). As minor impurities technical grade endosulfan may also contain up to 2 % endosulfan alcohol and 1 % endosulfan ether.



The technical product is a brownish crystalline solid and has a slight sulfur dioxide odour. Endosulfan is practically insoluble in water but soluble in most organic solvents. Log Kow values for technical endosulfan is 3.55 and 3.62, for α-endosulfan 3.83, for β-endosulfan 3.52, and for endosulfan sulfate 3.66. It is hydrolysed slowly by water, more rapidly by acids and bases. Decomposition is catalysed by iron, which it corrodes.

α - endosulfan β - endosulfan

Figure 1. Structure of α- and β-endosulfan.

Endosulfan is a non-systemic insecticide and acaricide with contact action. Formulations of endosulfan include emulsifiable concentrate, wettable powder, ultra-low volume (ULV) liquid, and smoke tablets. It is used in the control of sucking, chewing and boring insects and mites on a very wide range of crops, including fruit, vines, olives, vegetables, ornamentals, potatoes, cucurbits, cotton, tea, coffee, rice, cereals, maize, sorghum, oilseed crops, hops, hazels, sugar cane, tobacco, alfalfa, mushrooms, forestry, glasshouse crops, etc. In addition to its agricultural use, and its use in the control of the tsetse fly, endosulfan is used as a wood preservative and for the control of home and garden pests.

Endosulfan containing products are authorised for use in seven member states of the Community. But use of endosulfan within the EU has seen a steadily decrease lasting recent years. Almost 90 % of 490 tons/year used in 1999, were applied in Mediterranean parts of the EU (Umweltbundesamt, 2004). A draft Commission Decision has been notified by the European Communities to the WTO proposing not to include endosulfan in the positive Community list (Annex I of Directive 91/414/EEC) because it does not satisfy the minimum safety requirements in particular the environmental fate, eco-toxicological profile and the operators’ exposure risk. The foreseen date of entry into force is 1 August 2005 and Member States have to withdraw all existing authorisations for plant protection products containing endosulfan within 6 months from that date12.

1.2. Environmental fate

Endosulfan is released into the environment mainly as a result of its use as a pesticide. The compound partitions to the atmosphere and to soils and sediments. Endosulfan can be

12 Notification G/SPS/N/EEC/260 of 3 May 2005. http://www.wto.org/english/tratop_e/sps_e/sps_e.htm



transported over long distances in the atmosphere, but is relatively immobile in soils (ATSDR, 2000). Direct uptake from soil to plant as well as transport in the plant is negligible.

Both α- and β- endosulfan are fairly resistant to photo degradation, but the metabolites endosulfan sulfate and endosulfan diol (Figure 2) are susceptible to photolysis. The half-life of both isomers in water are estimated to be in the range of 4 to 7 days, but at low pH and anaerobic conditions it could be up to 5 months (ATSDR, 2000). In laboratory experiments with two soil types half time of 14C-endosulfan was estimated between 90 and 180 days based on 14CO2 production (Peres et al., 2004).

Figure 2. Chemical degradation of endosulfan in the environment (WHO, 1984).

In soil, the α-isomer has a shorter half-life (60 days) than the β-isomer (900 days). Endosulfan sulfate was found to be the major degradation product in soil and on plant surfaces. It is found to be more stable than the two endosulfan isomers, but the transport of all three compounds is slow in soil. Biodegradation in soil and water is dependent on climatic conditions and on the type of micro organisms present. In plants sprayed with endosulfan, initial residues on fruits and vegetables can vary from about 1 to 100 mg/kg; after 1 week, residues generally decrease to 20 % or less of the initial amount (WHO, 1984).

Due to its potential for long range transport, environmental persistence, bioconcentration in various aquatic organisms and ecotoxicity, there is agreement that endosulfan and its metabolite endosulfan sulfate meet the criteria for future inclusion into the list of persistent organic pollutants (POPs). However, unlike most other organochlorine pesticides of the Diels-



Alder class, such as chlordene, chlordane, heptachlor, heptachlorepoxide, aldrine and dieldrin, endosulfan has a less pronounced affinity to lipids. Consequently, biomagnification and accumulation of endosulfan, in terrestrial food chains, is unlikely to occur. Endosulfan is still in use in some countries.

1.3. Toxicological findings in laboratory animals and hazard assessment for humans

Endosulfan was evaluated by JMPR in 1998 (FAO/WHO, 1998). ATSDR published a toxicological profile for endosulfan in 2000 (ATSDR, 2000).

The neurotoxic effects of endosulfan are well documented in both humans and animals, and extensive research has been conducted in recent years aimed at elucidating its mechanism of neurotoxicity. Possible mechanisms of toxicity include (a) alteration of neurotransmitter levels in brain areas by affecting synthesis, degradation, and/or rates of release and reuptake, and/or (b) interference with the binding of neurotransmitter to their receptors. In addition to neurotoxicity, exposure to endosulfan has induced a wide array of effects in animals including liver and kidney toxicity, hematological effects, alterations in the immune system, and alterations in the reproductive organs of males. There are just a few studies on possible mechanisms of the effects on organ or systems other than the nervous system.

As summarized in 6.3. Metabolism, the biotransformation of endosulfan can give rise to a number of both polar and nonpolar metabolites. There is little and inconclusive information on whether the toxicological properties of endosulfan are due to the parent compound or to any of its metabolites.

The more lipophilic parent compound of endosulfan will be able to cross cell membranes more easily than its polar metabolites, accumulate to a greater extent, and therefore possibly be the most neurotoxic. Differential toxicity could also be related to differential affinity for the GABA receptor. What is known from oral acute lethality studies in rats and mice is that α-endosulfan is approximately 3 times more toxic than β-endosulfan (Dorough et al., 1978; Hoechst, 1975, 1990; Maier-Bode, 1968). In addition, in mice, the acute toxicity of endosulfan sulfate was comparable to that of α-endosulfan (Dorough et al., 1978). Also in mice, the metabolites endosulfan α-hydroxy ether, endosulfan lactone, and endosulfan ether had lethal doses 10 - 20 times higher than the α-or β-isomers; the lethal dose for endosulfandiol was two orders of magnitude higher than that of the α-or β-isomer (Dorough et al., 1978).

Most studies are carried out with technical products and if purity is given it is generally 97 - 99 % but purity as low as 91.4 % is also reported.

Acute toxicity

Acute exposure to high doses of endosulfan results in hyperactivity, muscle tremors, ataxia, and convulsions.



The LD50 of endosulfan varies widely depending on the route of administration, species, vehicle, and sex of the animal. Female rats are clearly more sensitive than male rats, and, on the basis of a single study, this sex difference appears to apply to mice also. The lowest oral LD50 value is 9.6 mg/kg b.w. in female Sprague-Dawley rats (Hoechst, 1990; Reno, 1975).

Long term toxicity

In a 78 week oral study with mice non-neoplastic changes were observed in both sexes in kidneys and sex organs. Based on these findings a NOAEL was identified for female mice at 3.9 mg/kg diet; equal to 0.58 mg/kg b.w./day (US National Cancer Institute, 1978).

In a corresponding 78 week oral study with rats given diets containing 220 mg/kg technical-grade endosulfan (purity, 98.8 %) or higher, non-neoplasic effects were seen in all dose groups and thus a NOAEL could not be established (US National Cancer Institute, 1978). In an other study groups of 50 five-to-six-week-old rats of both sexes were fed diets containing endosulfan (purity, 97.1 %) at concentrations of 0, 3, 7.5, 15, or 75 mg/kg diet, equal to 0, 0.1, 0.3, 0.6, and 2.9 mg/kg b.w./day for males and 0, 0.1, 0.4, 0.7, and 3.8 mg/kg b.w./day for females, for 104 weeks. Reductions in body weights and body-weight gains were observed in males and females at 75 mg/kg diet, but no clinical signs of poisoning were seen at any dose. No increase in mortality rates was observed in treated groups. Increased incidences of enlarged kidneys in females and of aneurysms, enlarged lumbar lymph nodes and marked progressive glomerulonephrosis in males were seen at 75 mg/kg diet. The NOAEL was 15 mg/kg diet, equal to 0.6 mg/kg b.w./day, on the basis of reduced body weights and pathological findings at higher doses (Ruckman et al., 1989; Gopinath and Cannon, 1990; Hack et al., 1995).

Human data as well as studies in animals did not provide unequivocal evidence of carcinogenicity for endosulfan (Hack et al., 1995; Hoechst, 1988, 1989). However, endosulfan promoted the development of altered hepatic foci in rats initiated with nitrosodiethylamine (Fransson-Steen et al., 1992). Although the mechanism of tumour promotion of endosulfan is not known, it has been suggested that it involves inhibition of cellular communication (Kenne et al., 1994).

JMPR (FAO/WHO, 1998) established an ADI of 0 - 0.006 mg/kg b.w. for technical endosulfan on the basis of the NOAEL of 0.6 mg/kg b.w./day in the two-year dietary study of toxicity in rats and a safety factor of 100. The ADI is supported by similar NOAEL values in the 78-week dietary study of toxicity in mice (NOAEL of 0.58 mg/kg b.w./day), a one-year dietary study of toxicity in dogs (NOAEL of 0.8 mg/kg b.w./day, see chapter 5.6 for further details), and a study of developmental toxicity in rats (NOAEL of 1.5 mg/kg b.w./day)

Endosulfan has been tested for genotoxicity in a wide range of assays. There was however no evidence on genotoxicity in most of these assays. In one assay for dominant lethal mutation in mice, late effects were observed at high doses (16.6 mg/kg b.w./day). The technical endosulfan used was reported to have a purity of 97.3 % (FAO/WHO, 1998, Pandey et al. 1990).



2. Methods of analysis

Analysis of endosulfan residues in food and feed samples should include detection of α- and β- endosulfan plus the major degradation product endosulfan sulfate. Currently, high resolution gas chromatography with electron capture detection (GC/ECD) or mass spectrometric detection (GC/MS) after extraction of samples with organic solvents, various clean-up steps to remove lipids and other possible co-extractives are the analytical methods of choice. These methods not only allow differentiation between the different isomers but also separate them from possible superimposing co-extractives. An efficient separation of the two endosulfan isomers and endosulfan sulfate from other interfering compounds, such as other organochlorine pesticides and polychlorinated biphenyls (PCBs) is especially important. In routine monitoring programmes it has therefore proven necessary to perform the gas chromatographic separation on two capillary columns of different polarity.

3. Statutory limits

Endosulfan is listed in the Annex to Directive 2002/32/EC of the European Parliament and of the Council of 7 May 2002 on undesirable substances in animal feed13 which replaces since 1 August 2003 Council Directive 1999/29/EC of 22 April 1999 on the undesirable substances and products in animal nutrition14. The maximum levels which apply to the sum of the α- and β- isomer and of endosulfan sulfate, expressed as endosulfan each pertain to a feedingstuff with a moisture content of 12 %. See also specific background.

Minimum time intervals between the last application and harvesting are prescribed in most countries and vary between 0 and 42 days, depending on the crop, type of formulation used, the mode of application, tolerances, and agronomic needs.

4. Occurrence in food and feed

Endosulfan is released to the environment mainly as the result of its use as a pesticide. It has been found at low levels in numerous food and feed samples.

Annually, some 40,000 – 50,000 food samples are analysed for, on average, 151 different pesticides within national monitoring programmes of 15 EU Member States and the three EFTA States Norway, Iceland and Liechtenstein in order to check the compliance of different food commodities with the corresponding pesticide maximum residue levels. The condensed results are regularly submitted to the Commission, which compiles and collates the data. Besides national monitoring programmes, the Commission recommends the participation of each Member State in a specific EU co-ordinated monitoring programme. These programmes which have existed since 1996 include the analysis of major components of the standard European diet of plant origin (so far: mandarins, pears, bananas, potatoes, oranges, peaches, carrots, spinach, cauliflower, wheat grains, peppers, melons, rice, cucumber, head cabbage, peas, apples, tomatoes, lettuce, strawberries, table grapes and nectarines) for an increasing 13 OJ L140, 30.5.2002, p. 10 14 OJ L 115, 4.5.1999, p. 32



number of pesticides. Endosulfan became part of the pesticide spectrum to check for in 1997. Between 1997 and 2002 a total of 35,152 food samples of plant origin have been analysed for endosulfan in the frame of these EU co-ordinated monitoring programmes. While 33,785 (96.1 %) samples contained no endosulfan residues, this pesticide could be determined in 1367 samples (3.9 %) below or at the MRLs. In 55 cases (0.16 %), the MRLs were exceeded. Endosulfan and endosulfan sulfate were found mainly on pepper, melons, strawberries and lettuce15.

Endosulfan levels in food can be reduced by food processing. For example, endosulfan can be removed by physical refinement treatment during oil refining using bleaching and deodorization (Riuz Mendez et al., 2005) or by peeling of apples (Rasmussen et al., 2003).

Thousands of feed samples are analysed annually in the Member States within the frame of official feed control, with the aim to check compliance with legal limits. As the Commission only requires the Member States to report their results as compliant and non-compliant, these condensed summaries give almost no information on actual levels in feed. Furthermore, it is often not specified which compounds are covered by the analytical method applied nor are the limits of detection reported. When comparing the summary reports it is often difficult to differentiate between numbers of individual analyses on the one hand and number of samples on the other hand. Concentration levels for individual substances analysed rather than condensed summaries for compound groups would be essential for a better understanding of the occurrence situation of undesirable substances in different feed materials and compound feeds as a prerequisite for a meaningful risk assessment and finally for a derivation of a possible temporal trend of the respective compounds in the feed chain.

Analysis of 104 feedingstuffs performed in 2003/2004 as part of official feed control in Germany revealed that endosulfan was detected in two samples only and then at a concentration of 7 µg/kg. In all other samples which included crops, maize, oil seeds, tubers, roots, mineral feed and compound feed for ruminants, pigs, poultry, horses and pets, α-, β-endosulfan and endosulfan sulfate could not be detected at a limit of detection of 1 µg/kg.

In the Czech Republic, 10 samples of fish meal have been analysed in 2004. The samples were analysed for α- and β-endosulfan. All results were below the limit of detection of 1 µg/kg. Data on endosulfan sulfate were not reported.

Data on fish feed (9 samples) provided by European Feed Manufacturers' Federation showed levels of 0.08 – 0.23 µg/kg for α-endosulfan, whereas the levels for β-endosulfan and endosulfan sulfate were < 0.1 µg/kg.

A survey of pesticide residues in animal feed ingredients has been conducted in the UK in 1998, where 151 samples of cereals (barley grain and malting, wheat grain, maize gluten and distillers), fodder (barley and wheat straw, grass and maize silage, sugar beet pulp) and beans (rapeseed, cotton seed, sunflower seed, cocoa meal, palm kernel and soy bean meal, copra)

15 European Commission, Food and Veterinary Office, pesticide annual reports

http://www.europa.eu.int/comm/food/fvo/specialreports/pesticides_index_en.htm



where analyzed for 28 different pesticides including endosulfan. None of these samples contained endosulfan at concentrations over the detection limit (50 µg/kg) (MAFF-UK, 1998).

5. Adverse effects on fish, livestock and pets, and exposure-response relationship

5.1. Introduction

The effect of any dose of endosulfan varies with the route of exposure and with the vehicle used. The lethal dose is lower if the insecticide is given in vegetable rather than in mineral oil, or as suspensions or dry powder (Humphreys, 1988). The sensitivity to endosulfan exposure varies with species, strain, age, gender and health status.

Acute intoxication with endosulfan is expressed through stimulation of the nervous system. The symptoms vary considerably but are predominantly neuromuscular. The onset of clinical signs may occur after a few minutes to days depending on the dose and route of exposure. Most animals show signs within 24 hours after exposure (Humphreys, 1988).

The signs of chronic endosulfan toxicity are principally similar to those of acute poisoning but develop more gradually, and tremors, convulsions, and depression may occur for weeks (Humphreys, 1988).

5.2. Fish

In a recently published study on Nile tilapia of about 140 g b.w., the effects of endosulfan at 100 and 500 µg/kg diet during 21 or 35 days exposure on thyroid hormone levels and metabolism were investigated (Coimbra et al., 2005). The plasma levels of T4 and of the inactive metabolite reverse T3 were decreased compared to levels in control fish, the effect was most prominent at the lowest dose. Furthermore, the hepatic deiodinase type I activity was reduced and the hepatic deiodinase type III (D3) was increased compared with controls. In the gills, D3 activity was increased in fish fed both levels of endosulfan. There was no clear dose-response pattern. It is therefore difficult to interpret the consequences of these effects on a long term; in addition, the authors claim that these might be adaptive responses.

In the common carp (on average 62.5 g b.w.) fed a diet containing technical endosulfan (thiodan, 35 % w./v.) at an endosulfan concentration of 0.5 µg/kg diet dry weight (0.015 µg/kg b.w.) for 5 weeks. No alterations in behaviour, feeding activity, growth and macroscopically overt signs of pathology upon dissection were observed. Cytological and ultrastructural alterations in hepatocytes and enterocytes were investigated by means of light and electron microscopy (Braunbeck and Appelbaum, 1999). The liver cells showed enlargement of the nucleolus, increased number and size of both Golgi fields and rough endoplasmic reticulum (ER) lamellae, as well as proliferation of peroxisomes and lysosomes, all together representing morphological equivalent of a general stimulation of hepatic metabolism. Furthermore, proliferation of smooth ER, glycogen and lipid depletion, invasion of phagocytic macrophages and accumulation of myelinated bodies in endothelial cells of



hepatic sinusoids were found. In the intestinal tract, lack of chylomicrons in the epithelial lining was observed, indicating a disturbance of absorption. These subtle biological changes were considered by the Panel as not to represent adverse effects.

Endosulfan is toxic to fish through exposure via water, and generally the LC50 values lie within the concentration range of 1 to 10 µg/L water (Goebel et al., 1982). The α-isomer was more toxic and the β-isomer less toxic, than technical grade endosulfan when examined in two freshwater fish species (Rao et al., 1980; Devi et al., 1981). Sulfur-free metabolites possess a significantly lower toxicity against fish with LC50 values in the range 1 to 10 mg/L (Knauf and Schulze, 1973). The effect of endosulfan in fish is temperature-dependent with decreased action at low temperatures, demonstrated in rainbow trout (Macek et al., 1969).

Short time endosulfan exposure of different fish species in sub lethal water concentrations (0.1 - 10 µg/L), have triggered increased swimming activity and raised blood glucose levels (Van Dyk et al., 1977; Gopal et al., 1980; Singh and Srivastava, 1981; Verma et al., 1983). Histopathological changes in the gills are reported after exposure to toxic water concentrations (Dalela et al., 1979). Histopathological changes in the liver and brain have been found in fish exposed to endosulfan contaminated water from insect spraying (Matthiessen and Roberts, 1982).

5.3. Ruminants

In an experimental study, groups of two steers were given endosulfan in the rations at doses 0.15, 1.1, 2.5 and 5.0 mg/kg b.w. After 60 days, no signs of toxicity were found in the pairs of steers receiving endosulfan at 0.15 and 1.1 mg/kg, One of the steers in the pairs receiving 2.5 mg/kg and 5.0 mg/kg showed toxic symptoms (muscle convulsions, excessive salivation, and incoordination) after 13 and 2 days, respectively, and both pairs were removed from the experiment (Bech et al., 1966). This indicates a NOAEL for clinical toxicity of 1.1 mg/kg b.w., and with a dry matter feed intake to body weight of approximately 3 % (Pond et al. 1995) this correspond to a no observed acute effect concentration of 40 mg/kg diet.

A study of endosulfan effects in goats orally dosed with 5 mg/kg b.w/day for 36 days revealed reduced body weight gain and depleted fat stores compared to controls. Other toxic symptoms were not reported (Amin and Abdalla, 1995).

There are several reports on endosulfan poisoning in cattle. In one case fatal poisoning appeared among cows fed peanut hay contaminated with endosulfan. Three cows died within 15 hours after the ingestion, and the dose was calculated to be approximately 30 mg of Thiodan (presumed to contain 35 % endosulfan)/kg b.w. (Terblanche and Minne, 1968). In another accidental case in cattle, Thiodan (also presumed to contain 35 % endosulfan) resulted in convulsions and death. The dose ingested was not determined, but residues were determined in liver (4.4 mg/kg w.w.), kidney (1.1 mg/kg w.w.) and muscle tissue (0.66 mg/kg w.w.) consisting predominantly of the metabolite endosulfan sulfate (Braun and Lobb, 1976).



Spraying of animals with accidental high endosulfan concentration (Thiodan emulsion of 0.12 % endosulfan) for treatment against ectoparasites, produced acute nervous symptoms in 50 of 250 cattle and 11 animals died (Thomson, 1966). Spraying or applying powder with the pesticide at the animal skin are the main causes to endosulfan poisoning in cattle and such cases are still occurring (Aslani, 1996; Kelch and Kerr, 1997; Mor and Ozmen, 2003).

5.4. Birds

In broiler chicken fed 30, 60 and 120 mg endosulfan/kg diet from age one day to eight weeks, a slight inhibitory effect on body weight gain relative to controls was found at all endosulfan levels, but no significant effect on feed consumption was revealed. Furthermore, increased serum glucose and decreased serum albumin was found in the chicken at all endosulfan levels. The hyperglycemia tended to be dose-related (Selvaraj et al., 2001a,b). The daily feed intake of broiler chicken after hatching is approximately 15 % relative to the body weight, reduced to approximately 10 % within few days (Pond et al., 1995), hence the levels in feed correspond to daily doses of 3 - 4.5, 6 - 9 and 12 - 18 mg/kg b.w., respectively.

Hudson et al. (1972) examined the effects of age of mallard ducks on their acute susceptibility to endosulfan. The oral LD50s of animals of 36 hours after birth, the age of 7 days, 30 days and 6 months were 28, 6.5, 7.9 and 34 mg/kg b.w., respectively.

Reported lethal feed concentrations (< 10-d LC50) of endosulfan in young mallard, ring-necked pheasant and bobwhite quail are 350, 175 and 100 mg/kg diet (WHO, 1984). Based on a daily feed intake of approximately 6 % relative to body weight, the feed concentrations correspond to approximately 20, 15 and 6 mg/kg b.w., respectively.

5.5. Rabbits

Mated New Zealand white rabbits were given endosulfan by gavage on days 6 - 28 of gestation at doses of 0.3, 0.7 or 1.8 mg/kg b.w. The highest dose was associated with signs of maternal toxicity that included noisy, rapid breathing, hyperactivity and convulsions, but no teratogenic or developmental effects. No clinical signs of toxicity in does or foetal effects were found at the two other dose levels. Thus, the NOAEL of endosulfan in rabbits was 0.7 mg/kg b.w./day (Dickie et al., 1981).

5.6. Dogs

Dogs dosed endosulfan 200 or 500 mg/kg b.w. as a single dose in gelatine capsules showed increased saliva formation, vomiting, and tonic and clonic cramps. The animals which did not vomit during these tests died (Maier-Bode, 1968). LD50 in dogs is reported 77 mg/kg (FAO/WHO, 1998)

Beagle dogs of both sexes were fed endosulfan at concentrations of 3, 10 and 30 mg/kg diet for one year, calculated by the authors to be 0.23, 0.77 and 2.3 mg/kg b.w./day. At the 30



mg/kg diet some animals had violent contractions of the abdominal muscles, and males at this dose had reduced body weight gains throughout the study in comparison with controls, and cholinesterase activity was increased in the brain. No other effects related to treatment were observed. In addition, one group was given a diet containing 30 - 60 mg/kg endosulfan, increasing in stages from 30 mg/kg for 54 days, to 45 mg/kg for 52 days, and 60 mg/kg for 19 - 40 days. These dogs were killed in extremis before the scheduled completion of the experiment due to a number of neurotoxic symptoms (FAO/WHO, 1998). The results from this study gives a NOAEL of 10 mg/kg feed, corresponding to 0.8 mg/kg b.w.

Mongrel dogs of both sexes were given endosulfan in gelatine capsules at feed levels 3, 10 and 30 mg/kg for one year. No clinical signs or treatment related effects on body weight gain, clinical chemical or haematological parameters, or gross or histopathological changes at these concentrations were noted (FAO/WHO, 1998). Middle sized adult dogs have daily maintenance requirement of dry feed of approximately 2 % relative to body weight (Pond et al., 1995), corresponding to 0.06, 0.2 and 0.6 mg/kg b.w./day.

6. Toxicokinetics and tissue disposition

6.1. Absorption

Several animal studies provided evidence of endosulfan absorption following oral exposure (Goebel et al., 1982; FAO/WHO, 1998; ATSDR, 2000). In metabolic studies with 14C-endosulfan in mice (Deema et al., 1966) approximately 65 % of the administered radioactivity was recovered from the excreta and tissues 24 hours after ingestion of a single dose (0.3 mg/animal), suggesting that gastrointestinal absorption occurred to a significant extent in this species. More than 90 % of a single oral dose of 14C-endosulfan (2 mg/kg b.w.) was absorbed in rats with a maximum plasma concentration occurring after 3 – 8 hours in males (0.25 µg/mL) and about 18 hours in females (0.18 µg/mL). The half-life in plasma was 75 hours in females. Males show a biphasic curve with an initial half-life of 8 hours followed by a half-live of 110 hours (FAO/WHO, 1998).

When 14C-endosulfan was administered as a single oral dose (0.3 mg/kg b.w.) to lactating sheep (Gorbach et al., 1968), blood radioactivity reached a maximum after 24 hours (0.064 µg/mL). The metabolic balance performed on day 22 suggested that absorption of endosulfan was > 42 % of the dose, based on radioactivity excreted in urine and milk.

Although no specific studies were carried out to determine the absorption of endosulfan in humans, residues of endosulfan were found in the fat, brain and kidney of a man who had ingested a single oral dose (260 mg/kg b.w.) of endosulfan. 150 minutes after ingestion, the levels of α- and β-endosulfan and endosulfan sulfate in blood were 644, 101 and 876 µg/L respectively (Boereboom et al., 1998). α-Endosulfan and endosulfan sulfate were also found in urine but at levels of 1.0 µg/L or lower. The patient died 91 hours after ingestion. Post mortem, the highest levels of α-endosulfan were found in adipose tissue and stomach content, 4.1 and 3.5 mg/kg respectively, whereas lower levels were found in brain and kidney, 0.08 and 0.06 mg/kg respectively. β-endosulfan was found in brain and stomach content at 0.07



and 1.4 mg/kg respectively and endosulfan sulfate was found in liver, brain and kidney at 3.0, 1.3 and 0.4 mg/kg respectively. β-endosulfan and endosulfan sulfate were not analysed in adipose tissue. Even if this report concerns an extreme case, it gives evidence of absorption, metabolism and distribution of endosulfan in humans.

6.2. Distribution

Studies using radiolabelled endosulfan administered to rat and mice indicate that the tissue concentration of residues of parent compound and metabolites were generally highest in the kidneys and liver and lower in other tissues, including fat. Male rats exposed daily for 60 days to 2.5 or 3.75 mg/kg/day of endosulfan containing α- and β-isomers in a ratio of 2:1 produced different disposition patterns for the two isomers (Ansari et al., 1984). For both doses, the concentrations of the α-isomer were as follows: kidney > epididymis > prostate ≈ spleen > testes > brain > liver. The β-isomer was found predominantly in seminal vesicle > epididymis > heart > prostate > spleen > liver. Overall, the greatest levels of both isomers were located in the kidney, seminal vesicle, and epididymis, with the liver having the least amount. Hoechst (1987) investigated the tissue residues in rats consuming 34 or 68 mg endosulfan/kg/day over 4 weeks. The predominant substances found in the liver were endosulfan sulfate and endosulfan lactone. Traces of α- and β-endosulfan were measured in the liver, whereas approximately 200 times more α-endosulfan than β-endosulfan was found in the kidney.

In lactating cows and sheep (Keller, 1959; Gorbach et al., 1968) residues were predominantly found in fat, kidney, and liver; all of the remaining tissues had considerably lower concentrations.

Because Spain is a main user of endosulfan within EU several reports focused on residues of endosulfan in humans from this country especially from southern Spain where extensive areas are devoted to intensive farming, including plastic greenhouse production.

In the mid-1990ties remarkable high levels of endosulfan residues were determined in the adipose tissue from 52 children from the Granada area in Spain, and showed the following results, based on a limit of quantification of 1 - 5 ng/g of lipid: α-endosulfan was detected in seven individuals: distribution, percentile 25, 50 and 75 were: 8.6, 58.9 and 105 ng/g of lipid. β-endosulfan was detected in three individuals with levels of 115, 2450 and 9060 ng/g of lipid. Endosulfan sulfate was found in one individual at 42.9 ng/g of lipid (Olea, personal communication).

In women of reproductive age the highest concentration of α-endosulfan, β-endosulfan and endosulfan sulfate were found in fat corresponding to 11 ± 86 µg/kg fat, 6.5 ± 20 µg/kg fat and 16 ± 93 µg/kg fat, respectively (Cerrillo et al., 2005).

6.3. Metabolism

Biotransformation in mammals is by oxidation, hydrolysis and subsequent conjugation of α- and β-endosulfan. The major portion of metabolites in the excreta and/or tissues consisted of



unidentified polar metabolites that could not be extracted from the matrix, whereas the non-polar metabolites including sulfate, diol, α-hydroxyether, and ether derivatives of endosulfan, represented only minor amounts (Dorough et al., 1978). In sheep, endosulfan sulfate was detected in the faeces and endosulfan diol and α-hydroxyether were detected in urine (Gorbach et al., 1968).

Of all the metabolites of endosulfan, the sulfate appears to be the one that accumulates, predominantly in the liver and kidneys. The β-isomer is more resistant to oxidation to endosulfan sulfate than the α-isomer and hence is more persistent in living organisms (Sutherland et al., 2004). Residues associated with endosulfan comprise both isomers and endosulfan sulfate, the latter being the major residue detected in animal tissues after exposure (Sutherland et al., 2004). This has important consequences in regard to the monitoring issue since endosulfan sulfate has equivalent mammalian toxicity to α-endosulfan (Goebel et al., 1982).

Endosulfan is slowly metabolised in fish and endosulfan sulfate is the main metabolite (Rao et al., 1981). Other metabolism data for fish dietary exposed to endosulfan were not identified.

6.4. Excretion

Elimination occurs mainly in the faeces and to lesser extent in urine, less that 15 % is retained after 5 days. In a study using rats treated with a single oral dose of 14C-endosulfan (2 mg/kg b.w.), Dorough et al. (1978) found that 82 and 72 % of the dose was found in faeces, whereas 12 and 22 % was excreted in urine for males and females, respectively.

Biliary excretion of radio labelled compounds in male rats given 1.2 mg/kg b.w. as a single dose was approximately 50 % for the α-isomer and 30 % for the ß-isomer over 48 hours. There appeared to be little enterohepatic circulation.

In sheep receiving a single dose of 14C-endosulfan (Gorbach et al., 1968), radiolabelled compounds were excreted mainly via the urine (41 %) and faeces (50 %). About half of the 50 % in faeces was unmetabolised endosulfan.

When treatment ceased after dietary administration of 14C-endosulfan for 14 days (Dorough et al., 1978), the estimated residues half-lives were approximately one week for kidney and three days in liver.

7. Carry over and tissue concentration

In pigs fed a diet containing 2 mg endosulfan/kg feed for up to 81 days, the compound was detected in fat at concentrations of 70, 90 and 40 µg/kg after 27, 54 and 81 days of treatment, respectively, suggesting that endosulfan does not bioaccumulate. 27 days after the exposure ceased concentrations in fat were below the limit of detection (Maier-Bode, 1966).

Groups of three lactating Holstein cows were given diets containing 0, 0.3, 3, or 30 mg/kg 14C-endosulfan for 30 days. Between days 7 and 29, the average concentrations in milk were



3.4, 40, and 462 µg/kg endosulfan in the three dose groups (Bowman, 1959). The concentrations of tissue residues found at the three doses were: liver, 0.35, 2.45, and 25.3 mg/kg; kidney, 0.05, 0.35, and 6.29 mg/kg; and omental fat, 0.07, 0.71, and 7.08 mg/kg (Keller, 1959).

After feeding 0.5 to 2 g technical endosulfan per day to dairy cows for 11 days, the parent compound endosulfan was not found in milk, but merely the oxidation product endosulfan sulfate (McCaskey and Liska, 1967). Braun and Lobb (1976) determined endosulfan concentrations in different tissues of a dairy herd acutely intoxicated by endosulfan. In post mortem samples from the carcass 1270 mg/kg was found in rumen content, whereas the concentrations in liver (4.2 mg/kg), kidneys (1.1 mg/kg) and muscle (0.6 mg/kg) were much lower. In animals which survived the concentrations in the milk decreased rapidly, with a biological half life of about 3.9 days.

Two lactating sheep were given a single oral dose of 0.3 mg/kg b.w. 14C-endosulfan and were killed after 40 days (Gorbach et al., 1968). The elimination of radio labelled compounds via milk during 17 days was 0.37 % and 1.82 % of the dose in the two sheep. Fat, kidney, and liver of the sheep contained 0.02 - 0.03 µg/g endosulfan; all of the remaining tissues had considerably lower concentrations. The total amount of radio labelled compounds found in organs and tissues accounted for less than 1 % of the administered dose.

In lactating goats receiving endosulfan in gelatine capsules at a dose of 1 mg/kg b.w. per day for 28 days, the levels of total residues (α- and β-endosulfan and endosulfan sulfate) were generally low, the highest being detected on the first day after cessation of treatment, with 0.29 mg/kg in kidney, 0.2 mg/kg in the gastrointestinal tract, 0.12 mg/kg in liver and 0.02 mg/kg in milk (Indraningsih et al., 1993). The concentration in the kidney was increased to 0.49 mg/kg one week after the treatment ceased, but no residues were detected 21 days after end of treatment. Endosulfan residues did not accumulate in the fat; the concentrations reached 0.06 mg/kg on day 1 after the end of treatment, but no residues were detected one week 8 after treatment.

Naqvi and Vaishnavi (1993) reviewed bioaccumulation factors for aquatic animals. For different organisms the reported bioconcentration factors were between 10 and 600. In an ATSDR review (ATSDR, 2000) maximum bioconcentration factors in aquatic systems are usually less than 3,000 and residues are eliminated within 2 weeks after transfer of fish to endosulfan-free water. The tests with 14C-labelled endosulfan revealed that fish are capable of forming water-soluble endosulfan metabolites in the liver. Analyses suggest that endosulfan diol is formed, which conjugated with glucuronic acid and is passed via bile to the faeces and excreted (Goebel et al., 1982).

Bargar et al. (2001a,b) studied the transfer of endosulfan injected in laying hens to the eggs. About 0.04 to 0.12 % of the dose injected was transferred to the eggs (Bargar et al., 2001b). During incubation of the fertilized eggs most of the endosulfan could be found in yolk and albumin. There seemed an excretion and reabsorption of endosulfan and metabolites into/from the allantoic fluid during the development of the embryo.



Endosulfan has been detected in breast milk of women environmentally exposed to a number of contaminants in rural Kazakhstan (Lutter et al., 1998) and in Spain (Cerillo et al., 2005), indicating that transfer to children can occur during lactation. No data on levels were reported in the Kazakhstan study, but in the Spanish study a mean endosulfan (α plus β) concentration of 11.38 ng/mL milk was found.

8. Human dietary exposure and comparison with ADI

The most important routes of exposure to endosulfan for the general population are ingestion of food and the use of tobacco products with endosulfan residues remaining after treatment (ATSDR, 2000). A total diet study performed between 1993 and 1996 in Canada revealed an average daily dietary intake for total endosulfan 23.8 ng/kg b.w. (Health Canada, 2003). In a recent Canadian study (Rawn et al., 2004) on a single location maximum intake of endosulfan could be observed in 5 - 11 year old children (0.03 µg/kg b.w./day). These values correspond with the results of an US study (0.05 µg/kg b.w./day) (Gunderson, 1995). In Hsinchu, Taiwan, the dietary intake of α- and β-endosulfan was studied from June 1996 to April 1997 (Doong et al., 1999). β-Endosulfan was not detected in any of the 14 different foods studied, including fruits, meats, seafood, and cereal, and α-endosulfan, by contrast, was found in 78 of 149 samples at an average concentration of 2.8 ng/g wet weight. Data on endosulfan sulfate were not reported in this study. Based on the average Taiwanese diet, the estimated daily intake of α-endosulfan was 0.62 µg/person. Converted to body weight this results at approximately 0.01 µg/kg b.w. (Doong et al., 1999). In Europe, the Czech Republic reported results of dietary intake for the sum of α-, β-endosulfan and endosulfan sulphate. Median of summary exposure to endosulfan in 1994 was 0.015 µg/kg b.w./day (Ruprich et al., 1995). Mean intake 0.003 µg/kg b.w./day has been reported in 2002 (Ruprich et al., 2003). 4136 samples representing all major food groups after processing were analysed during the period 1994 - 2002. Most frequently contaminated foods were offal (30 %) and fish and fish products (28 %) but concentrations were low (up to 15 µg/kg of sample).

CONCLUSIONS

Chemistry and environmental fate

• Once released into the environment, α- and β-endosulfan, which are the major constituents of the technical-grade pesticide endosulfan, can be broken down by hydrolysis, biodegradation. β-endosulfan is more persistent than the α- isomer. Endosulfan sulfate is the main degradation product of both isomers. It is equally toxic and more persistent in vivo than its parent compounds. Therefore, it is mandatory that analysis of endosulfan residues in feed and food includes the parent compounds α- and β-endosulfan as well as their major degradation product endosulfan sulfate.

Adverse effect in animals

• Fish show high sensitivity to endosulfan exposure via water. Oral exposure studies have shown effects on thyroxin level and thyroid hormone metabolism at dietary



concentration of 100 µg/kg (Nile tilapia), and ultrastructural alterations of the liver and intestinal tract at dietary concentration of 0.5 µg/kg (common carp). These effects were subtle, possibly adaptive, and not considered to represent adverse effects.

• The dominant toxic effect in mammals is stimulation and disturbance of the nervous system.

• A NOAEL for clinical toxicity of 1.1 mg/kg b.w. was found in young steers, fed endosulfan for 60 days, corresponding to a concentration in the diet of 40 mg/kg dry matter. This concentration in feed is 400 times higher than the current ML for the corresponding feed product.

• For chicken fed endosulfan for eight weeks a lowest observed adverse effect level of 30 mg/kg feed (LOAEL of 3 mg/kg b.w.) was found. This concentration in feed is 300 times higher than the current ML for the corresponding feed product.

• In dogs, orally dosed endosulfan for one year, a no observed adverse effect level of 10 mg/kg feed (NOAEL of 0.8 mg/kg b.w.) was found. This concentration in feed is 100 times higher than the current ML for the corresponding feed product.

Occurrence in feed and carry over

• Residues of endosulfan in feed are mostly reported by the Member States to the Commission as condensed overall summaries just giving the number of compliant and non compliant samples. Thus, primary data are not accessible, and as a consequence, detailed occurrence levels of endosulfan in feed are scarce.

• The limited data available on the occurrence of endosulfan and endosulfan sulfate in various feed categories, including fish feed, show only a limited number of samples containing residues (usually below 1 µg/kg product).

• Endosulfan does not significantly bioaccumulate in mammals.

• In living organisms, β-endosulfan is more persistent than α-endosulfan.

• Depending on species and duration of exposure, residues (parent compounds and endosulfan sulfate) are predominantly found in kidney, fat and liver. Transfer of residues to milk and eggs occurs to a limited extent.

• Based on the limited data on animal exposure via feed produced according to good agricultural practice, it is not likely that terrestrial animals will be exposed to levels that could cause toxic effects.

Human exposure

• Limited exposure data collected in the 1990s in Canada, United States and Taiwan show a mean daily dietary intake between 0.01 and 0.05 µg total endosulfan/kg body weight. Recent data on human endosulfan exposure in European adults are reported from the Czech Republic for the years 1994 - 2002. These show a mean daily dietary



intake between 0.003 and 0.015 µg total endosulfan/kg body weight. A similar dietary exposure can be assumed for other EU Member States based on the occurrence data of endosulfan in various food commodities measured in co-ordinated European monitoring programmes since 1997.

• Endosulfan does not significantly bioaccumulate in humans. During the latest WHO field study 27 human milk pools from 16 European and non-European countries were analysed for pesticides and showed no endosulfan contamination at a limit of detection of 1 µg/kg milk fat.

• The limited data available indicate that human dietary exposure to endosulfan is well below the ADI at 6 µg/kg b.w. set by JMPR in 1998.

Important gaps in the database

• Detailed data on residues of endosulfan and its metabolites in feedingstuffs and food of animal origin are scarce.

• Only limited information on oral toxicity of endosulfan exposure in fish and no data on laying hens are available.

RECOMMENDATION

• Most of the surveillance data from Member States are required by the European Commission to be reported as compliant or non-compliant. To allow for a better intake assessment it is necessary that the actual levels as well as the contaminant are reported.

• Studies on carry-over, accumulation and oral toxicity of endosulfan, especially in farmed fish and laying hens, should be performed.



REFERENCES

Amin, A.E. and Abdalla, G.A. 1995. Effects of endosulfan and amitraz on feedlot performance, carcass yield and meat quality characteristics of Nubian goats. Vet Hum Toxicol 37:113-115.

Ansari, R.A., Siddiqui, M.K.J. and Gupta, P.K. 1984. Toxicity of endosulfan: Distribution of alpha- and beta-isomers of racemic endosulfan following oral administration in rats. Toxicol Lett 21:29-33.

Aslani, M.R. 1996. Endosulfan toxicosis in calves. Vet Hum Toxicol 38:364. ATSDR (Agency for Toxic Substances and Disease Registry), 2000. Toxicological profile for

endosulfan. Atlanta GA. USA. Bargar, T.A., Scott, G.I. and Cobb, G.B. 2001a. Uptake and distribution of three PCB

conceners and endosulfan by developing white leghorn chicken embryos (Gallus domesticus). Arch Environ Contam Toxicol 41, 508 – 514.

Bargar, T.A., Scott, G.I. and Cobb, G.P. 2001b. Maternal transfer of contaminants: case study of the excretion of three polychlorinated biphenyl congeners and technical-grade endosulfan into eggs by white leghorn chickens (Gallus domesticus). Environ. Toxicol Chem 20, 61-67.

Beck, E.W., Johnson, J.C. Jr., Woodham, D.W., Leuck, D.B., Dawsey, L.H., Robbins, J.E. and Bowman, M.C. 1966. Residues of endosulfan in meat and milk of cattle fed treated forages. J Econ Entomol 59:1444-1450.

Boereboom, F.T., van Dijk. A., van Zoonen, P. and Meulenbelt, J. 1998. Nonaccidental endosulfan intoxication: A case report with toxicokinetic calculations and tissue concentrations. Clin Toxicol 36(4):345-352.

Bowman, J.S. 1959. Preliminary report: Subacute feeding – dairy cows. Unpublished report from Hazleton Laboratories, USA. Hoechst document No. A14205. Submitted to WHO by Hoechst Schering AgrEvo GmbH, Frankfurt-am-Main, Germany.

Braun, H.E. and Lobb, B.T. 1976. Residues in milk and organs in a dairy herd following acute endosulfan intoxication. Can J Anim Sci 56:373-376.

Braunbeck, T. and Appelbaum, S. 1999. Ultrastructural alterations in the liver and intestine af carp Cyprinus carpio induced orally by ultra-low doses of endosulfan. Dis Aquat Org 36: 183-200.

Cerrillo, I., Granada, A., Lopez-Espinosa, M.J., Olmos, B., Jimenez, M., Cano, A., Olea, N. and Fatima Olea-Serrano, M. 2005. Endosulfan and its metabolites in fertile women, placenta, cord blood, and human milk. Environ Res 98(2):233-9.

Coimbra, A.M., Reis-Henriques, M.A. and Darras, V.M. 2005. Circulating thyroid hormone levels and iodothyronine deiodinase activities in Nile tilapia (Oreochromis niloticus) following dietary exposure to endosulfan and Arochlor 1254. Comp Biochem Physiol 141C: 8-14.

Dalela, R.C., Bhatnagar, M.C., Tyagi, A.K. and Verma, S.R. 1979. Histological damage of gills in Channa gachua after acute and subacute exposure to endosulfan and rogor. Mikroskopie 35:301-307.



Deema, P., Thompson, E. and Ware, G.W. 1966. Metabolism, storage and excretion of C-14-endosulfan in the mouse. J Econ Entomol 59:546-550.

Devi, A.P., Rato, D.M.R., Tilak, K.S. and Murty, A.S. 1981. Relative toxicity of the technical grade material, isomers, and formulations of endosulfan to the fish Channa punctata. Bull Environ Contam Toxicol 27:239-243.

Dickie, S.M., McKenzie, K.M. and Rao, G.N. 1981. Teratology study with FMC 5462 in rabbits. Raltech Sci Serv, US Report No A23192 (unpublished study).

Doong, R.A., Lee, C.-Y. and Sun, Y.-C. 1999. Dietary intake and residues of organochlorine pesticides in food from Hsinchu, Taiwan. J AOAC Int 82:677-682.

Dorough, H.W., Huhtanen, K., Marshall, T.C. and Bryant, H.E. 1978. Fate of endosulfan in rats and toxicological considerations of apolar metabolites. Pest Biochem Physiol 8:241-252.

FAO/WHO (Food and Agriculture Organization/World Health Organization), 1998. Joint FAO/WHO Meeting on Pesticide Residues (JMPR). Endosulfan, part II, toxicology. http://www.inchem.org/documents/jmpr/jmpmono/v098pr08.htm.

Fransson-Steen R., Flodström, S. and Wärngård, L. 1992. The insecticide endosulfan and its two stereoisomerspromotethe growth of altered hepatic foci in rats. Carcinogenesis 13 (12):2299-2303.

Fürst, P. 2004. Personal communication. Goebel, H., Gorbach, S., Knauf, W., Rimpau, R.H. and Hüttenbach, H. 1982. Properties,

effects, residues, and analytics of the insecticide endosulfan. Res Rev 83:1-165. Gopal, K., Anand, M., Khanna, R.N. and Misra, D. 1980. Endosulfan induced changes in

blood glucose of catfish, Clarias batrachus. J Adv Zool 1:68-71. Gopinath, C. and Cannon, M.W.J. 1990. Photomicrographic addendum to histopathology

report No. Hst/289 Endosulfan, active ingredient technical (code: Hoe 002671 OI ZD97 0003) combined chronic toxicity/carcinogenicity study (104-week feeding in rats). Huntingdon Research Centre Ltd, Huntingdon, Cambridgeshire, United Kingdom. Unpublished Hoechst document No. A44604. Submitted to WHO by Hoechst Schering AgrEvo GmbH, Frankfurt-am-Main, Germany.

Gorbach, S.G., Christ, O.E., Kellner, H.-M., Kloss, G. and Borner, E. 1968. Metabolism of endosulfan in milk sheep. J Agric Food Chem 16:950-953.

Gunderson, E.L. 1995. FDA total diet study, July 1986 – April 1991, dietary intakes of pesticides, selected elements, and other chemicals. J AOAC Int 78:1353- 1363.

Hack, R., Ebert, E. and Leist, K.H. 1995. Chronic toxicity and carcinogenicity studies with the insecticide endosulfan in rats and mice. Food Chem Toxicol 33:941-950.

Health Canada, 2003. Average dietary intakes (ng/kg b.w./day) of pesticide residues for Canadians in different age-sex groups from the 1993 to 1996 Total Diet Study. http://www.hc-sc.gc.ca/food-aliment/cs-ipc/fr-ra/e_pesticide_intake_93-96.html



Hoechst, 1975. Beta-endosulfan purew (analysis GOE 1485): Acute oral toxicity in female SPF-Wistar rats. Hoechts Aktiengesellschaft, Frankfurt, Germany. Doc #A05270 (unpublished study).

Hoechst, 1987. Endosulfan - active ingredient technical (code HOE 02671 OI ZD97 0003): 30-Day feeding study in adult male Wistar rats. Hoechst Aktiengesellschaft, Frankfurt, Germany. Project no. 87.0129 (unpublished study).

Hoechst, 1988. Endosulfan - active ingredient technical (code HOE 02671 OI ZD97 0003): Carcinogenicity study in mice: 24-Month feeding study. Hoechst Aktiengesellschaft, Frankfurt, Germany. TOXN no. 83 0113 (unpublished study).

Hoechst, 1989. Endosulfan - active ingredient technical (code HOE 02671 OI ZD97 0003): Combined chronic toxicity/carcinogenicity study: 104-Week feeding in rats. Conducted for Hoechst Aktiengesellschaft, Frankfurt, Germany. Huntington Research Centre, Cambridgeshire, England. Project no. HST 289/881067 (unpublished study).

Hoechst, 1990. Summary and evaluation of the toxicity datafor endosulfan – substance technical (code HOE 002671) Hoechts Aktiengesellschaft, Frankfurt, Germany. Report no. 90 0848 (unpublished study).

Hudson, R.H., Tucker, R.K. and Haegele, M.A. 1972. Effect of age on sensitivity: acute oral toxicity of 14 pesticides to mallard ducks of several ages. Toxicol Appl Pharmacol 22(4):556-61.

Humphreys, D.J. 1988. Chlorinated hydrocarbon insecticides. In Humphreys, D.J. Veterinary Toxicology. 3. ed. Bailliere Tindall, London, p. 142-156.

Indraningsih, McSweeney, C.S. and Ladds, P.W. 1993. Residues of endosulfan in the tissues of lactating goats. Aust Vet J 70(2):59-62.

Kelch, W.J. and Kerr, L.A. 1997. Acute toxicosis in cattle sprayed with endosulfan. Vet Hum Toxicol 39:29-30.

Keller, J.G. 1959. Subacute feeding - dairy cows. Hazleton Laboratories, USA. Unpublished Hoechst document No. A14206. Submitted to WHO by Hoechst Schering AgrEvo GmbH, Frankfurt-am-Main, Germany.

Kenne, K., Fransson-Steen, R., Honkasalo, S. and Warngard, L. 1994. Two inhibitors of gap junctional intercellular communication, TPA and endosulfan: Different effects on phosphorylation of connexion 43 in the rat liver epitheliar cell line IAR 20. Carcinogenesis 15(6):1161-1165.

Knauf, W. and Scultze, E.-F. 1973. New findings on the toxicity of endosulfan and its metabolites to aqatic organisms. Meded Fac Lanbouwwet Rijksuniv Gent 38:717-732.

Lutter, C., Iyengar, V., Barnes, R., Chuvakova, T., Kazbekova, G. and Sharmanov, T. 1998. Breast milk contamination in Kazakhstan: implications for infant feeding. Chemosphere 37(9-12):1761-72.

Macek, K.J., Hutchinson, C. and Cope, O.B. 1969. Effects of temperature on the susceptibility of blugills and rainbow trout to selected pesticides. Bull Environ Contam Toxicol 4:174-183.



MAFF-UK (Ministry of Agriculture, Fisheries and Food in UK), 1998. Annual Report of the Working Party on Pesticides Residues: Supplement to The Pesticides Monitor 1999, MAFF Publications. p 18.

Maier-Bode, H. 1966. Investigations on the persistence of the insecticide endosulfan in the vegetable and animal organism. Pharmakologisches Institut der Rheinischen Friedrich Wilhelms Universität. Unpublished Hoechst document No. A4047. Submitted to WHO by Hoechst Schering AgrEvo GmbH, Frankfurt-am-Main, Germany.

Maier-Bode, H. 1968. Properties, effects, residues and analytics of the insecticide endosulfan. Res Rev 22:1-44.

Malisch, R., Kypke, K., van Leeuwen, R. and Moy, G. 2004. Unpublished data from the 3rd WHO human milk field study.

Matthiessen, P. and Roberts, R.J. 1982. Histopathological changes in the liver and brain of fish exposed to endosulfan insecticide during tsetse fly control operations in Botswana. J Fish Dis 5:153-159.

Mc Caskey, T.A. and Liska, B.J. 1967. Effect of milk processing methods on endosulfan, endosulfan sulfate, and chlordane residues in milk. J Dairy Sci 50:1991-1993.

Mor, F. and Ozmen, O. 2003. Acute endosulfan poisoning in cattle. Vet Hum Toxicol 45:323-324.

Naqvi, S.M. and Vaishnavi, C. 1993. Bioaccumulative potential and toxicity of endosulfan insecticide to non-target animals. Comp Biochem Physiol 105C:347 – 361.

Nath, G, and Dikshith, T.S.S. 1979. Endosulfan residues in rat tissues. Natl Acad Sci Lett 2:278-279.

Pandey, N., Gundevia, F., Prem, A.S. and Ray, P.K. 1990. Studies on the genotoxicity of endosulfan, and organochlorine insecticide, in mammalian germ cells. Mutat Res 242:1-7.

Peres, T.B., Papini, S., Marchetti, M. and Luchini, L.C. 2004. Dissipação de endossulfan em amostras de dois tipos de solos brasileiros tratadas em laboratório. Pesticidas: Revista de Ecotoxicologia e Meio Ambiente 14:11-18.

Pond, W.G., Church, D.C. and Pond, K.R. 1995. Basic animal nutrition and feeding. 4th ed. John Wiley & Sons, NewYork.

Rao, D.M.R., Devi, A.P. and Murty, A.S. 1980. Relative toxicity of of endosulfan, its isomers, and formulated products to the freshwater fish Labeo rohita. J Toxicol Environ Health 6:825-834.

Rao, D.M.R., Devi, A.P. and Murty, A.S. 1981. Toxicity and metabolism of endosulfan and its effect on oxygen consumption and total nitrogen excretion of the fish, Macrognathus aculeatum. Pestic Biochem Physiol 15:282-287.

Rasmussen, R.R, Poulsen, M.E. and Hansen, H.C.B. 2003. Distribution of multiple pesticide residues in apple segments after home processing. Food Additives and Contaminants 20(11):1044–1063.



Rawn, D.F.K., Cao, X.-L., Doucet, J., Davies, D.J., Sun, W.-F., Dabeka, R.W. and Newsome, W.H. 2004. Canadian total diet study in 1998: pesiticide levels in foods from whitehorse, yukon, canada, and corresponding dietary intake estimates. Food Add Contam 21:232-250.

Reno, F.E. 1975. Acute oral toxicity study in rats. Endosulfan technical. Unpublished final report, 18 December 1975, from Hazleton Laboratories America. Hoechst document No. A33732. Submitted to WHO by Hoechst Schering AgrEvo GmbH, Frankfurt-am-Main, Germany.

Riuz Mendez , M.V., Perez de la Rosa, I., Jimenez Marquez, A.and Ojeda, M.U. 2005. Elimination of pesticides in olive oil by refining using bleaching and deodorization. Food Add Contam 22:23-30.

Ruckman, S.A., Waterson, L.A., Crook, D., Gopinath, C., Majeed, S.K., Anderson, A. and Chanter, D.O. 1989. Endosulfan, active ingredient technical (code: Hoe 002671 OI ZD97 0003). Combined chronic toxicity/carcinogenicity study (104-week feeding study in rats). Huntingdon Research Centre, Huntingdon, Cambridgeshire, United Kingdom. Unpublished Hoechst document No.A40440. Submitted to WHO by Hoechst Schering AgrEvo GmbH, Frankfurt-am-Main, Germany.

Ruprich, J., Řehůřková, I., Steinhauserová, I. and Ostrý, V. 1995. Health impact of exposure to xenobiotics from food chains: alimentary diseases (1993) and dietary exposure (1994). National Institute of Public Health in Prague, 275 p., ISBN 80-900066-7-1.

Ruprich, J. Adamikova, V., Borkovcova, I., Bouskova, E., Dofkova, M., Karasek, K.,Karlikova, D., Karpiskova, K., Kolackova, I., Kopriva, V., Krajcovicova, R., Rajcovicova, R., Krbuskova, M., Markova, E., Ostry, V., Rehakova, J., Ehakova, J., Resova, D, Rehurkova, I., Slavikova, S., Skarkova, J. and Vokounova, S. 2003. Health impact of exposure to xenobiotics from food chain in 2002: reported alimentary diseases, bacteriological and mycological analyse of foods and dietary exposure of human beings. (in Czech). National Institute of Public Health in Prague, 2003. ISBN 80-7071-228-7.

Selvaraj, J., Balasubramaniam, G.A., Titus George, V. and Balachandran, C. 2001a. Effect of dietary endosulfan on the growth rate of broiler chicken. Indian Vet J 78:798-800.

Selvaraj, J., Balasubramaniam, G.A., Titus George, V. and Balachandran, C. 2001b. Studies on biochemical changes in endosulfan toxicity in broiler chicken. Indian Vet J 78:896-899.

Singh, N.N. and Srivastava, A.K. 1981. Effects of endosulfan on fish carbohydrate metabolism. Ecotoxicol Environ Saf 5:412-417.

Sutherland, T.D., Horne, I., Weir, K.M., Russell, R.J. and Oakeshott, J.G. 2004. Toxicity and residues of endosulfan isomers. Rev. Environ. Contam. Toxicol 183:99-113.

Terblanche, J. and Minne, J.A. 1968. Thiodan poisoning of cattle – a case report. Jl S Afr Vet Med Ass 39:91-92.

Thompson, G.E. 1966. Poisoning of cattle following accidental spraying with Thiodan. J S Afr Vet Med Ass 37:81-83.

Umweltbundesamt, 2004. Endosulfan - Draft Dossier prepared in support of a proposal of endosulfan to be considered as a candidate for inclusion in the UN-ECE LRTAP protocol



on persistent organic pollutants. German Federal Environment Agency – Umweltbundesamt, Berlin September 2004.

US National Cancer Institute, 1978. 78-week dietary study in Osborne-Mendel rats and B6C3F1 mice. NCI study No. NCI-CG-TR62, Technical Report Series No. 62, Bethesda, Maryland, USA.

Van Dyk, L.P. and Greeff, C.G. 1977. Endosulfan pollution of rivers and streams in the Loskop Dam cotton-growing area. Agrochemophysica 9:71.

Verma, S.R., Rani, S., Tonk, I.P. and Dalela, R.C. 1983. Pesticide-induced dysfunction in carbohydrate metabolism in three freshwater fishes. Environ Res 32:127-133.

WHO (World Health Organisation), 1984. Endosulfan. Environmental Health Criteria 40. International Programme on Chemical Safety. World Health Organization, Geneva, Switzerland.

SCIENTIFIC PANEL MEMBERS

Jan Alexander, Herman Autrup, Denis Bard, Angelo Carere, Lucio Guido Costa, Jean-Pierre Cravedi, Alessandro Di Domenico, Roberto Fanelli, Johanna Fink-Gremmels, John Gilbert, Philippe Grandjean, Niklas Johansson, Agneta Oskarsson, Andrew Renwick, Jirí Ruprich, Josef Schlatter, Greet Schoeters, Dieter Schrenk, Rolaf van Leeuwen, Philippe Verger.

ACKNOWLEDGEMENT

The Scientific Panel on Contaminants in the Food Chain wishes to thank Jan Alexander, Aksel Bernhoft, George Bories, Jean-Pierre Cravedi, Peter Fürst, Niklas Johansson and Hans Schenkel for the contributions to the draft opinion.

DOCUMENTATION PROVIDED TO EFSA

Submission of occurrence data Belgium, The Federal Agency for the Safety of the Food Chain, 2000-2004. Czech Republic, Central Institute for Testing and Supervising in Agriculture, 2004, and

National Institute of Public Health, 1994 - 2004. Germany, Chemisches Landes- und Staatliches, Veterinäruntersuchungsamt Münster, 2003

and 2004. European Feed Manufacturers' Federation.


Endosulfan does not persist in living beings

Health & Medicine

Transcript of Endosulfan does not persist in living beings