CARBENDAZIM, BENOMYL, AND THIOPHANATE-METHYL Summary Identity, physical and chemical properties, and analytical methods Sources of human and environmental exposure Environmental transport, distribution, and transformation Environmental levels and human exposure Effects on other organisms in the laboratory and the field Evaluation of effects on the environment Risk evaluation Aquatic environment Terrestrial environment Bibliography 1. Summary Carbendazim, benomyl, and thiophanate-methyl are evaluated together because benomyl and thiophanate-methyl are rapidly converted mainly to carbendazim in the environment. Benomyl has an aerobic half-life of 2 h in water and 19 h in soil; thiophanate-methyl has an aerobic half-life of less than one day in soil and two days in water. 2. Identity, physical and chemical properties, and analytical methods Carbendazim is an odourless, white crystalline solid with a melting-point of 302-307°C and a vapor pressure of 1 × 10-7 Pa (< 1 × 10-9 mbar) at 20°C; the technical grade has a melting-point of > 295°C. It has low water solubility, 8 mg/litre at 24°C and pH 7, and a low n-octanol:water partition coefficient (log Pow, about 1.5). Carbendazim is hydrolytically stable at pH 5-7 at ambient temperatures (22-25°C). It is capable of strong dust explosion, but is not flammable or auto-flammable. Analyses for residues and environmental contamination are performed by high-performance liquid chromatography (HPLC). Benomyl has a melting-point of 140°C, a vapour pressure of < 5.0 × 10-6 Pa at 25°C, a water solubility of 3.6 mg/litre at pH 5, and a log Pow value of 23.4-32.0. HPLC and immunoassays have been used to analyse for benomyl. Thiophanate-methyl has a melting-point of 177°C, a vapour pressure of 1.30 × 10-5 kPa, a water solubility of 21.8 mg/litre at 25°C, and a log Pow value of 25-34. 3. Sources of human and environmental exposure Carbendazim has been produced commercially since 1970. It is a systemic fungicide with a broad spectrum of activity on many economically important phytopathogens of the Ascomycetes, Deuteromycetes, and Basidiomycetes groups. It is used on agricultural crops (e.g. cereals, rice, grapes, cucurbits, tomatoes, pome fruits, stone fruits, and strawberries) and horticultural plants and in forestry and home gardening. Pest resistance has been seen, e.g. apple scab, eyespot, and Botrytis In order to combat resistance, carbendazim is combined with other fungicides with different modes of action. Benomyl and thiophanate-methyl degrade primarily to methyl-2- benzimidazole carbamate (carbendazim), which is also fungicidal. The mode of action of carbendazim involves interference in the biosynthesis of DNA during fungal cell division. As benomyl and thiophanate-methyl have a highly specific mode of action, resistant strains of fungal pathogens have been developed. The application rates and the number of treatments allowed per crop vary with the chemical (see Table 1). Overall, the rates are 0.18-0.60 kg/ha per application for carbendazim, 0.14-1.12 kg/ha for benomyl, and 0.22-2.13 kg/ha for thiophanate-methyl. Table 1. Uses and application rates (kg active ingredient/ha) and numbers of treatments allowed Commodity Carbendazim Benomyl Thiophanate-methyl Application No. of treatments Application No. of treatments Application No. of treatments rate rate rate Apples 0.3-0.6 4 0.45 3 1.35 > 9 Vineyards 0.28-0.56 4 0.45 4 Stone fruits 0.3-0.6 2 0.45 3 1.35-2.13 4 Curcurbits 0.30 2 0.30 3 0.22-0.50 ? Tomatoes 0.60 4 0.30 3 Rice 1.12 2 Wheat 0.25 1 (cereals) 1.12 1.5 0.9 1 Bananas 0.15 6 Citrus 1.12 2 Caneberries 0.42 5 Celery 0.28 6 Conifers 0.56 5 Peanuts 0.14 12 0.50 Unspecified Soya bean 1.12 1 0.5-0.9 2 Strawberries 0.60 2 0.56 5 0.7-0.9 Unspecified Beans 060 1 1.70-2.24 2 0.90-1.53 2 Nut crops Pecans 0.28-0.56 > 4 0.50-0.90 ? Almonds 0.56-0.84 2 1.35-1.80 2 Sugar beets 0.21-0.28 ? 0.29-0.38 ? Sugar cane 0.21-0.28 ? 0.38 ? 4. Environmental transport, distribution, and transformation Carbendazim has a half-life to hydrolysis of 25 weeks. In the environment, it has half-lives of 6-12 months on bare soil, 3-6 months on turf, and 1-2 months in a water-sediment system under aerobic conditions and 25 months under anaerobic conditions. In aerobic river sediment containing water, thiophanate-methyl degraded with a half-life of 15-20 days. Residues of carbendazim and its metabolites are strongly bound or incorporated into soil organic matter. In laboratory studies of soil containing labelled carbendazim, the molecules remained in the topsoil. Residues of carbendazim and its metabolites that are bound or incorporated into soil are probably due to the imidazole ring of the molecule, as the Koc is relatively low. Benomyl is hydrolysed rapidly, in < 2 h at pH 5, 7, and 9; it is rapidly degraded in water, with half-lives of < 4 h in sun or darkness, and on soil, at less than four days on silt loam soil. In a field study, the degradation half-life was 5.2 h in light and 5.7 h in the dark, indicating that photolysis is not a significant degradation pathway but that hydrolysis is the controlling factor. Under anaerobic conditions, 98% of carbendazim residues partition into sediment after seven days. The half-life of carbendazim was 743 days, and after one year 36% of the radiolabel was bound to sediment. Under aerobic conditions, the half-life in non-sterile sediment was 61 days. After 30 days, 22% of the radiolabelled carbendazim was bound to sediments, and < 1% of the amount applied was evolved as carbon dioxide. Anaerobic degradation in soil and water is slower than aerobic degradation. Carbendazim was susceptible to photolysis in natural daylight and was broken down to dimethyloxalate, quinidine, and mono- and dicarbomethoxyguanidine. No photolysis products were detected in extracts of leaves of treated corn plants after exposure to sunlight for 18 h. After aerobic incubation, 3-10 mg/kg carbendazim had disappeared within 250 days, with only 5-13% remaining. After 224 days, 57% was in the form of non-extractable residue; after 250 days, 4-8% was converted to 2-amine-1 H-benzimidazole and 30-40% to carbon dioxide. When carbendazim was applied directly to two German water- sediment systems, the label disappeared rapidly from water, with half-lives of 31 days in Rhine River water and 22 days in pond water. The average half-life of carbendazim in an aquatic system was about 26 days. In the total soil-water system, 98.3% carbendazim was eliminated from Rhine River water and 95.9% from pond water after 91 days. In contrast to English and Mississippi sediments, formation of 14C-carbon dioxide was extensive, amounting to 61.2% in Rhine River water and 40.3% in pond water after 91 days of incubation. Two other benomyl degradates, dibutylurea and 1-butylisocyanate, are formed under hot, humid conditions in greenhouses. These two compounds are being studied for phytotoxicity to vascular plants. Carbendazim, thiophanate-methyl, and benomyl are absorbed by sprouts, leaves, and roots and are translocated within the transpiration system in the plants xylem. They inhibit the development of fungal germ tubes, the formation of appressoria, and the growth of mycelia. Their fungitoxic action is based on blockage of nuclear division during mitosis and destabilization of fungal cell structures. Carbendazim, thiophanate-methyl, and benomyl persist on leaf surfaces and in leaf litter. The conversion of benomyl to carbendazim on leaf surfaces varies with the leaf surface, light intensity, and other environmental factors. About 50% of a 50% wettable powder formulation of benomyl remained on apple leaves after five days. Bioconcentration of carbendazim in fish is not expected to be significant: > 94% of residue was lost from whole fish, viscera, and muscle during a 14-day depuration phase. The bioconcentration factors for bluegill sunfish ( Lepomis microlophus) were 27 (0.018 mg/litre) and 23 (0.17 mg/litre). After exposure of rainbow trout ( Salmo gairdneri) and bluegill sunfish to 45 µg/litre for 96 h, rainbow trout had the highest uptake rate constant (1.78 per hour) and bioconcentration factor (159). Channel catfish ( Ictalurus punctatus) accumulated less residue (0.44 µg/g) but died after 48 h of exposure. The bioconcentration factors in whole fish were 23-27, and radiolabel was concentrated in the viscera, the peak viscera bioconcentration factors being 380-460; very little radiolabel was concentrated in muscle (bioconcentration factor, < 4) or the remaining carcass (< 7). When the fish were placed in clean water, > 94% of the peak level of radiolabel was lost from the whole fish, viscera, and muscle after two weeks; the rate of loss from the carcass was 77-82% at that time. 5. Environmental levels and human exposure Carbendazim residues are expected to persist on leaf surfaces and in leaf litter and to increase with each successive application. When a 50% wettable powder benomyl fungicide was applied to apple trees in three successive treatments at a rate of 1.7 kg/ha, benomyl residues on foliage had fallen by 50% on day 5 after application, whereas those of carbendazim had doubled and increased with each successive benomyl application. A similar pattern for residues of thiophanate-methyl can be expected. Thiophanate-methyl residues were monitored in water and in crayfish after application of Topsin M(R), a 70% wettable powder formulation, twice to rice fields. No thiophanate-methyl residues were detected in crayfish tails (detection limit, 0.05 ppm), and no adverse effects on the crayfish were observed. In rice water effluent sampled three to four weeks after the second application of 0.85 kg/ha, no thiophanate-methyl residues were found; carbendazim was detected at 0.01-0.006 ppm in the field, 0.01-0.007 in the drainage ditch, and 0.004-0.005 ppm at the receiving stream. In long-term studies of field dissipation, fallow and cropped fields received two treatments of Topsin M(R) at 1.57 kg/ha per application, and soil samples were collected for one year at depths of 0-10, 10-20, and 20-30 cm. The fields were tilled by standard practice for the area. The residues of thiophanate-methyl and carbendazim were low, even on the day of application. Thiophanate-methyl readily degraded to carbendazim in all soils, and the carbendazim residues declined to < 0.1 ppm in almost all plots after one to three months. Thiophanate-methyl was not detectable in soil sections of < 10 cm, and, after four months, carbendazim was not detected at > 0.02 ppm in the 20-30-cm soil section. Soil binding of carbendazim increased with the organic matter content of the soil. Dissipation of thiophanate- methyl did not occur below 0-10 cm, and carbendazim was essentially undetectable in the 20-30-cm layer. Dissipation of carbendazim in water was rapid. 6. Effects on other organisms in the laboratory and the field Carbendazim is highly acutely toxic to channel catfish (LC50, 0.007-> 0.56 mg/litre), aquatic invertebrates (0.087-0.46 mg/litre), mysid shrimp (0.098 mg/litre), and rainbow trout (0.1-> 1.8 mg/litre), and moderately to slightly toxic to bluegill sunfish (> 3.20- 55 mg/litre) in the laboratory. In a study of the early life stage of rainbow trout, the maximum acceptable toxicant concentration (MATC) was 0.019 mg/litre. In a two-generation study of reproductive toxicity in Daphnia magna, the MATC was 0.0040 mg/litre. In a study of the life cycle of Mysidopsis bahia, the MATC was 0.0354 mg/litre. The toxicity of carbendazim in green algae appears to be species-dependent, with median effective concentration (EC50) values ranging from 0.34 mg/litre for Chlorella pyrenoidosa to 419 mg/litre for Scenedesmus subspicatus. Carbendazim was algicidal to Raphidocellis subcapitata, with an EC50 of 1.6 mg/litre and a no-observed-effect concentration (NOEC) of 0.5 mg/litre. Significant inhibition of the Egyptian soil fungi Aspergillus sp. occurred when carbendazim was applied at the recommended field application rate; however, the nitrification activities of Nitrosomonas sp. and Nitrobacter agilis were not inhibited in suspension cultures with up to 100 mg/litre carbendazim. The mud ditch organisms Escherichia coli and Salmonella typhimurium were not adversely affected at up to 1000 mg/litre. Results of laboratory studies with terrestrial organisms are summarized in Table 2. The tests with natural arthropod enemies of pests, hover flies, ladybirds, and predatory mites, were conducted with only one dose of benomyl, so that the concentrations that had an effect cannot be calculated. Concentrations well below most recommended field rates, however, affected reproduction in at least one mite and one insect species. Several field studies of earthworms, in which benomyl or carbendazim were applied at rates near or below the maximal recommended field rates, are summarized in EHC 148 and EHC 149. In most of the reports, effects were recorded on numbers, biomass, cast production, and/or removal of litter. In a study in apple orchards, benomyl was applied at 0.28 kg/ha five to seven times a year for three years and thiophanate-methyl at 0.78 kg/ha seven times a year for two years. The numbers and biomass of the earthworm populations were seriously diminished, Lumbricus terrestris and Allolobophora chlorotica being the most affected. Populations of other earthworm species recovered within two years after spraying ended. Earthworm populations adjacent to the orchards were unaffected, perhaps due to the immobility of benomyl in soil (Stringer & Lyons, 1974). Field studies with soil and litter microorganisms show only transient or no effects of benomyl and carbendazim when applied at recommended field rates. Pronounced but transient effects (< 40 days) were observed on soil fungi by Abdel-Fattah et al. (1982), and Torstensson & Wessén (1984) found pronounced effects only in sandy soils at 2 kg/ha. No field studies were available on other non-target terrestrial organisms. Carbendazim has low toxicity for birds, with an acute oral LD50 value of > 2250 mg/kg bw for bobwhite quail. In five-day dietary studies, the LC50 values were > 10 000 mg/kg diet for mallard ducks and bobwhite quail. Owing to its low octanol:water partition coefficient, bioaccumulation is expected to be minimal. In a 90-day study of reproduction in Japanese quail, the NOEL for carbendazim was 160 mg/kg diet (mean daily intake, 20 mg/kg bw); the reproductive NOEC was 400 ppm, equivalent to a mean daily intake of about 50 mg/kg bw. Benomyl has low toxicity for birds, with an acute oral LD50 value of > 2250 mg/kg bw for bobwhite quail. In five-day dietary studies, the LC50 values were > 10 000 mg/kg bw for mallard ducks and bobwhite quail and > 5000 for Japanese quail. Because of its low octanol:water partition coefficient, its bioaccumulation is expected to be low. No studies are available on the effects of benomyl on avian reproduction. Thiophanate-methyl has low, toxicity for birds, with acute oral LD50 values of > 4640 mg/kg diet for bobwhite quail and mallard ducks. In five-day dietary studies, the LC50 values were > 10 000 mg/kg for mallard ducks and bobwhite quail. Owing to its low octanol:water partition coefficient, bioaccumulation is expected to be low. In 24-week studies of reproduction, the no-effect concentrations in diets were 500 mg/kg for bobwhite quail and 103 mg/kg for mallard ducks. Table 2. Toxicity of carbendazim, benomyl, and thiophanate-methyl to terrestrial organisms Organism Study type Exposure End-point, result (nominal concentration of active ingredient) Carbendazim Soil microorganisms Oxygen consumption 15 days EC50 > 100 mg/litre Nitrobacter agilis and Nitrification and NOEC > 100 mg/litre Nitrosomonas spp. witrification Escherichia coli and NOEC > 1000 mg/litre Salmonella typhimurium Aspergillis Growth 5 and 40 Significant inhibition Worms Compost worm (Eisenia andrei) Acute, artificial soil 3 weeks LC50 5.7 mg/kg Compost worm (Eisenia andrei) Reproduction, artificial soil 8 weeks EC50 2.9 mg/kg soil; NOEC, Insects 0.6 mg/kg Springtail (Folsomia candida) Reproduction, artificial soil 4 weeks EC50 > 1000 mg/kg soil Honey bee (Apis mellifera) Contact 48 h LD50 > 50 µg/bee Carabid beetle (Ptesrostichus melanarius) Contact, spray, soil/sand 6 days NOEC > 1250 g/ha Birds Dietary 5 days LC50 > 15.59-2250 mg/kg feed Reproduction 12 weeks NOEC 160-400 mg/kg diet Benomyl Soil microorganisms Dehydrogenase inhibition 28 days NOEC 6 mg/kg Worms Compost worm (Eisena foetida) Acute contact 14 days LC50 10.48 mg/kg Insects Hover fly (Syrphus corollae) Reproduction, glass plate, 11 days at 49% reduction larvae 3 µg/cm2 Ladybird (Coccinella septempunctata) Reproduction, glass plate, 3 weeks at No effect larvae 0.65 µg/cm2 Predatory mite (Typhlodromus pyri) Reproduction, glass plate, 19 days at 38% reduction protonymph 0.69 µg/cm2 Honey bee (Apis mellifera) Contact LD50 > 10 µg/bee Table 2. (cont'd) Organism Study type Exposure End-point, result (nominal concentration of active ingredient) Birds Japanese quail (Coturnix Dietary 5 days LC50 > 5000 mg/kg feed coturnix japonica) Redwing blackbird (Agelaius Oral Acute LD50 100 mg/kg bw phoeniceus) Thiophanate-methyl Soil microorganisms CO2 production 14 days NOEC > 30 mg/kg bw Nitrification 42 days NOEC > 30 mg/kg bw Heterotrophic N fixation 7 days NOEC > 5 mg/kg bw Worms Compost worm (Eisenia foetida) Acute, artificial soil 14 days LC50 20.8 mg/kg soil Insects Honey bee (Apis mellifera) Contact 48 h LD50 > 100 µg/bee Birds Bobwhite quail (Colinus virginanus) Dietary 5 days NOEC 4640-> 10 000 mg/kg bw LC50 > 10 000 mg/kg feed Mallard duck (Anas platyrhyncos) Acute oral 8 days LD50 > 4640 mg/kg bw From Van Gestel et al. (1992); Kühner (1992); Pietrzik (1992); Decker (1993) 7. Evaluation of effects on the environment Benomyl, carbendazim, and thiophanate-methyl are broad-spectrum systemic fungicides belonging to the benzimidazole group and are used on a wide variety of crops. Because of the development of resistance, benzimidazole fungicides are usually alternated with other compounds with different modes of action. Formulations include wettable powders, water-dispersible granules, flowable concentrates, dusts, and granules. Benomyl and thiophanate-methyl that enter the environment are converted to carbendazim, which can be regarded as the environmentally relevant compound. The half-lives are 2-19 h for benomyl and three to four days for thiophanate-methyl. The carbendazim formed decomposes in the environment with a half-life of months under aerobic and anaerobic conditions in soil and water. Carbendazim partitions from water to soil and sediment. It binds to the mineral component of the soil, probably through the imidazole ring. Adsorption is strong and carbendazim does not leach through the soil profile, despite its low Kow. No contamination of ground-water can be expected, as confirmed by field monitoring of well-water. Abiotic degradation is considered to be a minor route of degradation for carbendazim; microorganisms, predominantly bacteria, represent the major route of loss. Only moderate bioaccumulation of carbendazim was seen in laboratory studies at constant concentrations. There is rapid depuration on transfer to clean water. No significant bioaccumulation of carbendazim is expected in the field. Benomyl had no effect on soil bacterial populations in the laboratory. In studies in greenhouses and the field, application rates of up to 89.6 kg/ha had little effect on soil microbial populations. Limited studies suggest that thiophanate-methyl does not adversely inhibit soil-nitrifying bacteria. Field studies confirm these findings, even though transient effects on soil fungi have been observed. The NOEC for a green algae was 0.5 mg/litre. Carbendazim is highly to highly acutely toxic to fish, aquatic invertebrates, and mysid shrimp, with LC50 values of 0.007 mg/litre for fish, 0.087 mg/litre for aquatic invertebrates, and 0.098 mg/litre for shrimp. The MATC was 0.019 mg/litre for rainbow trout, 0.004 mg/litre for Daphnia magna, and 0.035 mg/litre for mysid shrimp. Field application rates of carbendazim are not expected to pose an acute hazard to non-target mammalian wildlife species. Carbendazim, thiophanate-methyl, and benomyl have low acute toxicity for birds, with dietary LC50 values > 5000 mg/kg. Field application rates of carbendazim are not expected to pose a hazard to birds. Risk assessment (a) Aquatic environment A simple screening model (Generic Expected Environmental Concentration -- Environmental Protection Agency/Office of Pesticide Products) for worst-case scenarios was used to estimate the predicted expected concentration (PEC) of carbendazim in aquatic systems after application of 0.56 kg/ha of carbendazim to vineyards in four treatments at 14-day intervals. The following parameters are used in the calculations: Soil Koc, 250; water solubility, 8 ppm; present spray drift, 5; depth of soil incorporation, 0; soil aerobic metabolic half-life, 180 days; aquatic aerobic half life, 61 days; longest hydrolysis half-life, 175 days, and photolysis half-life, stable. The concentrations found with this model in a water body 2 m deep were: peak, 56 µg/litre; four-day concentration, 54 µg/litre; 21-day concentration, 48 µg/litre; and 56-day concentration, 38 µg/litre. The model is basically the same as that used in the United Kingdom and the Netherlands. Toxicity exposure ratios (TERs) were calculated, as shown in Table 3, which indicate that a risk to all aquatic organisms, on either an acute or a chronic basis, is at least present and often large. Reduced bioavailability owing to adsorption to sediment would reduce this apparent risk. Contamination of surface waters by benomyl, carbendazim, and thiophanate-methyl must be avoided to prevent toxic effects on aquatic organisms. (b) Terrestrial environment The predicted environmental concentration (PEC) used for evaluating acute effects is based on one application of 0.6 kg/ha, of which 100% reaches both the canopy and the soil. The soil PEC for chronic effects is based on four successive applications of 0.6 kg/ha, as on vines. As the degradation rate of carbendazim in soil is of the order of months and carbendazim is not mobile, no degradation is assumed. A further assumption was that the compound is dispersed into the top 5 cm of a soil with a density of 1.4 g/cm3. The summary of acute and chronic TERs (Table 4) indicates a 'large' risk to earthworms in the soil. The risk to honey bees is regarded as low. The large risk to earthworms is confirmed by field studies; no such studies are available for arthropods. Table 3. Estimates of acute and chronic risk for aquatic organisms after application of carbendazim to vineyards Time course Organism PEC Toxicity End-point TER Risk classification (µg/litre) (µg/litre) Acute Invertebrate 56 87 LC50 1.55 Present Acute Fish 56 7 LC50 0.125 Large Acute Shrimp 56 98 LC50 1.75 Present Chronic Invertebrate 48 27 NOEC 0.56 Large Chronic Fish 48 3 NOEC 0.06 Very large Chronic Shrimp 45 35 NOEC 0.78 Large PEC, predicted environmental concentration; TER, toxicity:exposure ratio; NOEC, no-observed-effect concentration Table 4. Estimates of acute and chronic risk for terrestrial organisms after application of carbendazim to vineyards Time Organism PEC Toxicity End-point TER Risk classification course Acute Compost worm 0.86 mg/kg soil 5.7 mg/kg LC50 6.6 Present (Eisenia foetida) Acute Honey bee 6 µg/cm3 > 50 µg/bee LC50 12a Low (Apis mellifera) Chronic Compost worm 3.43 mg/kg 0.6 mg/kg soil NOEC for 0.17 Large (Eisenia foetida) reproduction Chronic Springtail 3.43 mg/kg > 1000 mg/kg Reproduction > 290 Negligible (Folsomia candida) Chronic Predatory mite 6 µg/cm3 38% reduction Reproduction - - (Typhlodromus pyri) at 0.69 µg/cm3 PEC, predicted environmental concentration; TER, toxicity:exposure ratio a Hazard ratio estimate from toxicity per bee Bibliography Abdel-Fattah, H.M., Abdel-Kader, M. & Hamida, S. (1982) Effect of Bavistin, Cotoran, and Curacron on Egyptian soil fungi. Mycopathologica, 80, 101-106. Ammon, H.U. (1985) Worm toxicity tests using Tubifex tibifex. 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See Also: Toxicological Abbreviations Carbendazim (EHC 149, 1993) Carbendazim (HSG 82, 1993) Carbendazim (ICSC) Carbendazim (WHO Pesticide Residues Series 3) Carbendazim (Pesticide residues in food: 1976 evaluations) Carbendazim (Pesticide residues in food: 1977 evaluations) Carbendazim (Pesticide residues in food: 1978 evaluations) Carbendazim (Pesticide residues in food: 1983 evaluations) Carbendazim (Pesticide residues in food: 1985 evaluations Part II Toxicology) Carbendazim (Pesticide residues in food: 1995 evaluations Part II Toxicological & Environmental) Carbendazim (JMPR Evaluations 2005 Part II Toxicological)