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Ethylenethiourea in air and in urine as an indicator of exposure to ethylenebisdithiocarbamate fungicides.

Ethylenethiourea (ETU) is a ubiquitous impurity of the ethylenebisdithiocarbamate (EBDC) fungicides widely used in agriculture and forestry. In the present study, ETU was used as a measure of the exposure to EBDC on potato farms and in pine nurseries during the application of EBDC fungicides and the weeding of the sprayed vegetation. Biological and hygienic monitoring was carried out through the analysis of ETU in the breathing zone and the urine of exposed workers. Even if the concentrations of ETU in the ambient air of pine nurseries exceeded those of potato farms, the concentrations of ETU in the urine of potato farmers exceeded those of pine nursery workers. This result may have been due better protective equipment in the pine nurseries. The excretion rate was 6-10 ng/h during the first 60 h after the cessation of exposure, and it diminished thereafter to 0.2 ng/h over a 22-d observation period.

Adult↗

Indicators of mancozeb exposure in relation to thyroid cancer and neural tube defects in farmers' families.

OBJECTIVES: The fungicide mancozeb has been suspected of causing thyroid cancer and neural tube defects. The aim of the study was to investigate associations of indicators of mancozeb exposure with thyroid cancer and neural tube defects in farmers' families. METHODS: National registers in Norway, identifying 105 403 female and 131 243 male farmers, born in 1925-1971, and their 300 805 children available for analyses, born in 1952-1991, were cross-linked with national agricultural censuses, 1969-1989, and the population register. Neural tube defects (ICD-8 740-742) at birth among the 102 703 children conceived between May 1973 and April 1991 were identified in the medical birth register. Likewise thyroid cancer (ICD-7 194) was identified in the cancer register through 2000. Data on farm production and fungal forecasts (humid and temperate weather conditions) in 1973-1990, obtained from agricultural censuses and meteorological measurement stations, respectively, served as the mancozeb exposure indicators. The adjusted rate ratio or prevalence ratio (PR) estimates with 95% confidence intervals (95% CI) were computed in a Poisson regression analysis. RESULTS: Neural tube defects (131 cases, prevalence 12.8/10 000 births) was moderately associated with potato cultivation (PR 1.6, 95% CI 1.1-2.3) and paternal work of > 500 hours/year (PR 1.6, 95% CI 1.1-2.5). Altogether 319 thyroid cancer cases were identified: 141 in female farmers (incidence 10.2/100 000 person-years), 79 in male farmers (incidence 3.2/100 000 person-years), and 99 in offspring (female and male incidence 3.4 and 0.6/100 000 person-years, respectively). Mancozeb exposure was not associated with thyroid cancer. CONCLUSIONS: A moderate association seems to exist between mancozeb exposure and neural tube defects, but not between mancozeb exposure and thyroid cancer.

Agriculture↗

Assessment of toxicological effects of mancozeb in male rats after chronic exposure.

Mancozeb, an ethylenebisdithiocarbamate fungicide was administered orally to male rats at doses 0, 500, 1000 and 1500 mg/kg/day for 90, 180 and 360 days produced dose dependent signs of poisoning, loss in body weight gain and mortality. However the signs of toxicity and mortality were more pronounced initially at 0-90 days as compared to 90-360 days of treatment period. A significant increase in the relative weight of liver and slight decrease in the kidney weight were observed in animals exposed to mancozeb (1000 and 1500 mg/kg/day) for 180 and 360 days associated with pathomorphological changes in liver, brain and kidney. Mancozeb has produced significant enzymatic changes in the activities of aspartate aminotransferase (ASAT), alanine aminotransferase (ALAT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and acetylcholinesterase (AChE) throughout the period of study in a dose dependent manner. The alterations in the activity of enzymes associated with pathomorphological changes suggest that the chronic exposure of mancozeb produced significant toxicological effects in rats.

Acetylcholinesterase↗

Transplacental carcinogenic potential of the carbamate fungicide mancozeb.

We evaluated the effects of mancozeb (Dithane M4-5), a protective carbamate fungicide, on transplacental carcinogenesis in Swiss albino mice. Mancozeb, a polymeric complex of ethylene bis (dithiocarbamate) manganese with zinc salt, is reported to possess carcinogenic and cocarcinogenic activity in various tumor models. In the present study, pregnant Swiss albino mice were administered mancozeb intraperitoneally on the 14th day of gestation. The first filial generation (F1 progeny) was promoted with a well-known tumor promoter 12-o-tetradecanoyl phorbol-13-acetate (TPA). The results revealed a significantly high tumor incidence (72%) in the F1 progeny of the animals initiated with mancozeb or a well known carcinogen 7,12-dimethyl benzanthracene (DMBA) and promoted with TPA in comparison to animals that were either from mothers given only the vehicle (DMSO) and promoted with TPA in F1 progeny or not promoted with TPA in F1 progeny. No significantly higher tumor incidence was observed in any other experimental groups. These results suggest that mancozeb or its metabolites are capable of crossing the placental barrier and can exert DNA damage and tumor initiating consequences in the fetal cells that, after promotion with TPA, get converted into neoplastic cells.

9,10-Dimethyl-1,2-benzanthracene↗

A rapid method for the determination of manganese ethylenebisdithiocarbamate and its residues on grains.

A method has been developed, based on the periodate oxidation of the manganese in the coordination product of zinc ion and manganese ethylene bisdithiocarbamate to permanganic acid, to determine DithaneM-45. The relationship between the intensity of color and the concentration of manganese obeys Beer's law up to 100 mug at 540 nm. The method can be easily applied to the determination of the active ingredient in formulations and also for residue analysis. The method is sensitive to 0.6 mug and can be used to determine 6 mug fungicide at the 20 g sample level.

Edible Grain↗

The effectiveness of ecologically acceptable ways of protection of field-grown tomato (Lycopersicon lycopersicum (L.) Karsten) from tomato late blight (Phytophthora infestans (Mont.) de Bary) in extreme weather conditions.

During the extremely wet vegetation season in 1999 the efficiency of protection of two cultivars of field grown tomato (Lycopersicon lycopersicum (L.) Karsten), 'Pick Rite' and 'GO 101', against tomato late blight (Phytophthora infestans (Mont.) de Bary) using three ecologically acceptable growing techniques and agrochemical measures (growing on black plastic mulch with no fungicides used, spraying with Dithane M-45 (mancozeb), spraying with Cuprablau-Z (Cu-hydroxide) and control) have been studied. It was found out that under such extreme weather conditions none of these measures can ensure an adequate protection of tomato. The highest yield of healthy fruits (0.30 kg per plant) was obtained after a threefold spraying with Dithane M-45 (mancozeb), the highest percentage of non infected fruits (38.1%) after a twofold spraying with Cuprablau-Z (Cu-hydroxide). The black plastic mulch, used with the intention to reduce the plant contact with the potential inoculum in the soil proved to be ineffective.

Agriculture↗

One-generation reproduction toxicity study of mancozeb and lead acetate.

The reproduction toxicity of lead acetate and 80% mancozeb containing fungicide formulation (Dithane M-45) were studied on rats. The lead acetate was applied in diet at the following dose groups: Control-1,000-5,000-10,000 mg/kg. Three treatment and a control groups were applied, 4,500 mg/kg Dithane M-45 was administered in all the dose levels simultaneously in diet. The basis of the method was the OECD Guideline for Testing of Chemicals No. 415. Clinical symptoms and mortality were not found in the parent generation. The body weight of female animals diminished significantly before the pregnancy period. This tendency was also seen on males after the combination treatment. Remarkable body weight growth of female animals was observed during lactation period at the two high dose levels. Diminished body weight data of offsprings were measured after treatment at the end of the lactation period. The histological examination showed a general tubulonephrosis in the trial. Summing up, it can be established the administration of fungicide Dithane M-45 did not increase the toxicity of lead acetate.

Animals↗

Rinsing and management of pesticides' containers.

In order to reduce the effects on the environment, it is necessary to improve the management of pesticides' containers. Usually, users burn or bury empty containers. These methods, even though decreasing must be avoided or even forbidden. Since 1996, empty containers are systematically collected in Belgium and are specifically removed by the firm Phytofar Recover created by the Belgian Federation of pesticides' manufacturers. Since the beginning, the recovery rate (percentage of containers recovered compared with the containers sold) goes on increasing to exceed 85% in 2001. These action and results are a world first (more than 500 tons of empty containers are collected yearly). Once collected, empty containers are subjected to the European Policy about toxic wastes since they contained dangerous products. Their removal must follow a specific removal process by incineration at very high temperature (> 1200 degrees C) with a specific filtration of the smoke. The treatment cost is high and reaches 2 Euros per kg of container. If the container is rinsed and the residue does not exceed 1000, 10,000 or 30,000 mg per kg of container (depending on the dangerousness of product: very toxic, corrosive or toxic), it will be considered as domestic waste and will therefore follow a much more economical energy production process. The study aims at determining the quantities of residue contained in empty containers and the parameters reducing the rinsing efficiency: the formulation (EC, WP, WG), the container's size, packaging's type (plastic container or paper bag), the rinsing technique. Almost 150 tests and analyses of residue have been carried out. A manual rinsing procedure has been set up in order to meet the standards about residue. Rinsing three times with an average volume of water (20 to 30%) allows to reach the lowest residue level. As bags containing powder (WG or WP) container not be rinsed, it is necessary to empty them completely. It is however difficult to reach the 1000 ppm residue limit.

Belgium↗

Toxicity of a mancozeb containing fungicide formulation and CU-sulphate to chicken embryos after administration as single compounds or in combination.

Environmental pollution of metal modelled by copper-sulphate and a 80% mancozeb containing fungicide formulation (Dithane M-45) were studied on chicken embryos after administration as a single compounds or in combination. The test materials were injected into the air-chamber in a volume of 0.1 ml/egg on day 12 of incubation. The concentration of copper-sulphate was 0.01%. The applied concentration of Dithane M-45 fungicide formulation was 0.2%. Evaluation was done on day 19 of the hatching period. The combined administration of copper-sulphate and the fungicide formulation did not cause a significant reduction in body weight as compared to the control data and the results from individual toxicity study of the test materials. After the combined administration of copper-sulphate and the fungicide formulation the rate of embryomortality was 40%. The incidence of developmental anomalies were sporadic. Light microscopic findings exhibited a degenerative change in the liver tissue of combined administration group. Activities of GPT and GOT enzymes increased markedly in the combined administration group. In summary, it can be established that the interaction of copper-sulphate and an 80% mancozeb containing fungicide formulation (Dithane M-45) caused higher embryomortality with respect to the test of individual toxicity of copper-sulphate and fungicide in our study.

Alanine Transaminase↗

Synergism in mixtures of cymoxanil and mancozeb on Phytophthora infestans in vivo.

The preventive activity of 1:8 mixture of cymoxanil and mancozeb against Phytophthora infestans was higher than that of either the two single ingredients or the other nine mixtures. The synergistic interaction existed (synergy ratio 2.01) between the two at the mixing ratio of 1:8, whereas additive interaction (synergy ratios ranged from 0.73 to 1.34) existed at the mixing ratios ranging from 1:1 to 1:7, from 1:9 to 1:10, 1:8 was the optimal ratio. The preventive activity of 1:8 mixture was higher than the curative and the eradicative. In addition, the eradicative synergism of inhibiting sporangia production on lesions was stronger than the eradicative synergism of inhibiting lesion extension and suppressing infection of sporangia, and than the curative synergism of inhibiting lesion sporulation on detached potato leaflets.

Acetamides↗

The effect of poisoning with Dithane M-45 on oxygen uptake and energy balance in adult rats.

Adult white rats were kept on natural diet containing various doses of Dithane M-45 (Mn and Zn ethylene-bisdithiocarbamate). Oxygen uptake (VO2), metabolic rate and energy balance were determined at intervals of several days. Oxygen uptake and metabolic rate were not significantly changed in animals receiving Dithane M-45 in concentrations of 5 and 50 ppm but they were significantly decreased in the animals kept on the diet containing the preparation in concentrations of 500, 2000 and 5000 ppm. The authors conclude that changes in body weight of animals after poisoning with Dithane M-45 are due, both to reduced supply and requirements for energy.

Animals↗

Pyrazolo[3,4-d][1,2,3]triazole-1-carboxamides and 5-alkylaminopyrazolo[3,4-d]oxazoles: synthesis and evaluation of the in vitro antifungal activity.

A series of N-alkyl-N'-(4-diazo-5-pyrazolyl)-ureas (4) was thermally and photochemically converted into pyrazolo [3,4-d][1,2,3]triazole derivatives (5,6) and 5-alkylaminopyrazolo[3,4-d]oxazoles (7) respectively. The products were tested for in vitro antifungal activity against Fusarium culmorum, Botrytis cinerea, Phoma betae, Pythium ultimum, Sclerotinia minor and Rhizoctonia solani. The MIC and ED50 values of compound (6) against some of the test fungi were comparable to those of the reference fungicides iprodione and mancozeb.

Aminoimidazole Carboxamide↗

Comparing two methods of examination in the interaction study of a pesticide and a heavy metal.

The use of pesticides in field application involves the risk of poisoning wild animals. The reproduction period of pheasant takes place at the same time as the spraying time of pesticides, which justifies, that we evaluate in a point of the ecotoxicologic view the influence of the pesticide on progressive avian embryo. The most frequent technical way is injecting the exam stuffs to the some part of the embryonated eggs under the bird teratological trials. The advantage of this method is that it can be injected in a correct measured dose into the optional part of eggs. The disadvantage of this method is that it can't model properly the influence on the environment. If adverse effect of the embraced chemical substance on the embryo is experienced under the study, it will be necessary to use an immersion treatment. This procedure shows only the possible indirect influence of the pesticide on the embryo but it can suitably model its influence in plant protection practice. Treatment was done on day 12 of incubation. Applied concentration of heavy element (Cd sulphate) was 0.01% and the concentration of pesticide (Dithane M-45) was 0.2%. Evaluation was done on day 19 of incubation. Injection treatment: the simultaneous administration of Cd sulphate and the 80% mancozeb containing fungicide formulation on day 12 of incubation did not result in a significant decrease in the average body weight of embryos compared to neither the control nor the pesticide individually treated group. At the same time the body weight of embryos significantly decreased because of combined administration as compared to the Cd sulphate treated group. The embryo mortality and the incidence of developmental anomalies markedly increased after the simultaneous administration. Immersion treatment: the combined administration of Cd sulphate and the mancozeb containing fungicide formulation on day 12 of incubation did not result in a significant decrease in the average body mass of embryos compared to neither the control nor the individually treated groups. The number of embryo mortality was very high after the simultaneous administration. The incidence of developmental anomalies was sporadic.

Animals↗

Embryonic toxicity of a mancozeb containing fungicide formulation and Cu-sulphate in pheasant after individual or combined administration.

Cu-sulphate and a 80% mancozeb containing fungicide formulation (Dithane M-45) were studied in pheasants after administration as single compounds or in combination. The test materials were injected directly into the air-chamber of eggs on day 12 of the hatching period and evaulation was carried out on day 23 of incubation. The pheasant embryos were examined for the following: rate of embryo mortality, body mass, type of developmental anomalies, light microscopic examination. After the administration of copper-sulphate on day 12 of incubation, the average body weight of pheasant embryos significantly did not decrease as compared to the control. The embryonic mortality was 68%. After the administration of a mancozeb containing fungicide formulation (Dithane M-45) on day 12 of incubation, the average body weight of embryos did not decrease as compared to the control. The embryonic mortality was 50%. After the individual administration of pesticide, the incidence of developmental anomalies was very high. After the combined administration of copper-sulphate and the 80% mancozeb containing fungicide formulation (Dithane M-45) on day 12 of incubation the embryonic mortality markedly increased. The rate of embryo mortality was 93%. We did not find any degenerative change in the liver tissue in either the control group or the treated groups by light microscopic examination. In summary, it can be established that the simultaneous administration of Cu-sulphate and mancozeb containing fungicide formulation caused high toxicity compared with the individual toxicity of test materials.

Animals↗

Toxicity of a mancozeb containing formulation and Cd-sulphate to chicken embryos after administration as single compounds or in combination.

Environmental pollution of metal modelled by cadmium-sulphate and a 80% mancozeb containing fungicide formulation (Dithane M-45) were studied on chicken embryos after administration as a single compounds or in combination. The test materials were injected into the air-chamber in a volume of 0.1 ml/egg on day 0 of incubation. The concentration of cadmium-sulphate was 0.01%. The applied concentration of Dithane M-45 fungicide was 0.2%. Evaluation was done on day 19 of the hatching period. The individual administration of cadmium-sulphate and the 80% mancozeb containing fungicide formulation did not cause a significant reduction in body weight as compared to the control data. Embryonic mortality increased at all individual treated groups and reached almost a 35% rate. After the individual administration of pesticide, the number of chicken embryos with developmental anomalies did not differ markedly from the control. After the combined administration of cadmium-sulphate and the 80% mancozeb containing fungicide formulation (Dithane M-45) on day 0 of the hatching period embryonic mortality markedly increased. 88% of the treated embryos were dead. Results from the combined administration of cadmium-sulphate and an 80% mancozeb containing fungicide formulation (Dithane M-45) caused higher embryomortality with respect to individual toxicity test of cadmium-sulphate and fungicide in our study.

Animals↗

Field studies to determine mancozeb based spray programmes with minimal impact on predatory mites in European vine cultivation.

Mancozeb is an ethylene bisdithiocarbamate (EBDC) fungicide with contact activity against a wide range of economically important fungal diseases. Its multi-site mode of action means that to date there have been no recorded incidences of resistance developing despite many years of use on high risk diseases. One such disease, Grape downy mildew (Plasmopara viticola) has developed resistance to a number of important oomycete specific fungicides following their introduction onto the market. The role of Mancozeb either as a mixing or alternation partner in helping to manage these resistance situations remains critically important. Historical use patterns for mancozeb in tree and vine crops involved many applications of product at high use rates. Although this gave excellent disease control, a negative impact on predatory mites has been reported by researchers. This has lead to the development of mancozeb spray programmes in vines and other crops with a much reduced impact on predatory mites. A range of field studies was conducted in France, Germany, Italy, Portugal and Spain where either 2 or 4 applications of mancozeb containing products were made per season at different spray timings. These trials covered the representative range of uses, agronomic practices, mite species and geographical locations in Europe. In this paper findings from ten field studies in five different vine growing regions in Europe indicated that two to four applications of mancozeb at 1.6 kg a.i./ha as part of a spray programme caused minimal impact on naturally occurring populations of predatory mites which in turn was compatible with Integrated Pest Management programmes and the conservation of predatory mites.

Animals↗

Persistence and degradation of metalaxyl, mancozeb fungicides and its metabolite ethylenethiourea in soils.

The degradation pattern of metalaxyl, mancozeb and its metabolite ethylenethiourea (ETU) residues indicated a close correspondence to first order exponential degradation kinetics in soils. Degradation of fungicides in soils was predominantly biological as well as chemical in nature. Slower degradation ofmetalaxyl was noticed in the soils and their half-life values were higher than mancozeb and ETU as evident by wide range of half-life values from 41.24 to 165.11 days. In case of metalaxyl, Hiriyur soil was found to be superior in degrading the metalaxyl. Lower persistence of mancozeb and ETU was observed in soils resulting in rapid rate of degradation at smaller half-life values as compared to metalaxyl indicating the faster degradation of mancozeb and ETU. In mancozeb applied soils, the ETU formation was increased up to 30 days of incubation and thereafter it declined. Amongsoils, degradation of either mancozeb or ETU is not influenced by soil types. However, mancozeb persistence was higher in Hiriyur soils than Chettalli and Bangalore soils.

Agriculture↗