The chronic toxicity of methiocarb to grackles,doves, and quail and reproductive effects in quail

Summary Methiocarb (4-methylthio-3,5-xylylN-methyl carbamate, Mesurol, Bay 37344), a bird repellent, was fed in concentrations of 100 to 1,000 ppm to common grackles (Quiscalus quiscula), mourning doves (Zenaida macroura.), and breeding pairs of coturnix quail (Coturnix coturnix) to investigate the possibility of cumulative intoxication. Although aversion to treated diets was readily apparent in most of the tests, the 28-to 30-day median lethal concentration (LC₅₀) was determined to be >100 ppm for grackles, 630 ppm (95% confidence limits, 480–830 ppm) for doves, and >1,000 ppm for coturnix quail. Methiocarb appeared to be noncumulative when measured by an index of chronicity: birds consumed several LD₅₀ doses during a day's feeding, and when deaths occurred, they appeared to be due to acute intoxication. Egg production and live chick production were not affected in coturnix fed 100 ppm but were reduced at 316 and 1,000 ppm.     

Studies on combined effects of organophosphates and heavy metals in birds. I. Plasma and brain cholinesterase in coturnix quail fed methyl mercury and orally dosed with parathion

Summary We found that mercury potentiated the toxicity and biochemical effects of parathion. Male Coturnix quail (Coturnix coturnix japonica) were fed a sublethal concentration of morsodren (4 ppm as methyl mercury) for 18 weeks. This resulted in an accumulation of 21.0 ppm of mercury in the liver and 8.4 ppm in the carcass. Birds fed clean feed and those fed morsodren-treated feed were orally dosed with 2, 4, 6, 8, and 10 mg/kg parathion, and their 48-h survival times compared. The computed LD₅₀ was 5.86 mg/kg in birds not fed morsodren and 4.24 in those fed the heavy metal. When challenged with a sublethal, oral dose of parathion (1.0 mg/kg), morsodren-fed birds exhibited significantly greater inhibition of plasma and brain cholinesterase activity than controls dosed with parathion. Brain cholinesterase activity was inhibited 41% in morsodren-fed birds and 26% in clean-fed birds dosed with parathion, which suggested that the increase in parathion toxicity in the presence of morsodren was directly related to the inhibition of brain cholinesterase.     

The prolonged exposure of Japanese quail to carbaryl and its effects on growth and reproductive parameters

Summary Carbaryl fed to Japanese quail over a 14 week period resulted in decreased body weight, increased relative brain, liver and kidney weights in one or both sexes at levels of 900 ppm or higher. Significant differences were not found in the F₁ birds at the same levels after 6 weeks. It appears that the time of exposure is critical and that parental diet is not. There were no significant differences in egg production, hatchability or fertility although trends toward reduction of these parameters were developing at the 900 and 1200 ppm levels.Paper No.4984 of the Journal Series of the North Carolina Agricultural Experiment Station, Raleigh, NC 27607.The use of trade names in this publication does not imply endorsement by the North Carolina Agricultural Experiment Station of the products named, nor criticism of similar ones not mentioned.Senior author on study leave of absence from the Environmental Protection Agency, Health Effects Research Laboratory, Research Triangle Park, NC 27711.     

The chronic toxicity of 3-chloro-4-methyl benzamine HCl to birds

3-Chloro-4-methyl benzamine HCl (DRC-1339), an avian toxicant, was fed to five species of birds for periods up to 120 days. The 30-day LC₅₀ of uniformly treated feed for starlings was 4.7 ppm and the 90-day LC₅₀ was 1.0 ppm. The 28-day LC₅₀ for coturnix was 18 ppm. The 30-day LC₅₀ for pigeons was less than 100 ppm. Pheasants fed diets containing 2% DRC-1339 baits diluted to a rate of 286 ppm of DRC-1339 died within 22 days. Bobwhite quail fed similar diets suffered some mortality at levels as low as 2.9 ppm, but most survived 10 times this dosage level for the 120-day test period. Application of the Kenaga Index of Chronicity, resulted in the conclusion that DRC-1339 was cumulatively toxic to birds.Reproduction in coturnix was adversely affected by treatments at 10 ppm of DRC-1339 and above. Reproduction in pigeons was adversely affected by a treatment of 25 ppm. In coturnix, DRC-1339 caused an increased incidence of egg breakage and decreased both egg and live chick production. In pigeons, DRC-1339 caused an increase in the proportion of infertile eggs. Reproductive ability of first generation offspring was not affected when parent coturnix and pigeons were fed DRC-1339.These data emphasize the need for care in the use of DRC-1339. The bait should be used only as registered and care exercised in storage and disposal of unused baits to avoid poisoning of nontarget species.     

Barbiturate potentiation in mercury poisoning

Summary Barbiturate potentiation was observed in Japanese quail fed dietary levels of 4, 21, and 24 ppm Hg as methyl mercuric chloride. Gross symptoms of mercury poisoning were not observed until after BP was observed. After the initial increase in BP time was observed in the birds receiving 4 and 12 ppm mercury, there followed a plateau in response until those birds receiving 24 ppm Hg began to show gross symptoms of toxicity and the, BP time markedly rose again. Pronounced BP persisted 7 weeks after removal of mercury from the diet reflecting the long biological half-life of mercury as the methyl derivative. Selenium effectively prevented BP after 7 days exposure to toxic levels of methyl mercury chloride. Therefore, results suggest that early toxic effects of mercury can be observed readily by BP and that this environmental contaminant may influence drug activity or increase the sensitivity of the nervous system to sodium pentobarbital.Scientific Article No.A1993, Contribution No.4935 of the Maryland Agricultural Experiment Station.     

Subchronic lead exposure, immunotoxicology and increased disease resistance in Japanese quail (Corturnix coturnix japonica)

The present study investigated the effects of lead (Pb) on immune responses in quail (Coturnix coturnix japonica) and the pathological impact of exposure to an infectious agent (E. coli O2). Fifty-four, 4-week-old quail were exposed to lead acetate in drinking water at 5 or 50 ppm. All birds were vaccinated with Newcastle Vaccine (NDV) during the third week of contaminant (Pb) exposure. In the fourth week, several arms of the immune response were tested using the T cell based phytohemagglutinin (PHA) skin test, the B cell mediated antibody response to NVD, and the chemiluminescence assay measuring innate immunity. Immunohistochemistry (IHC) was used to determine the expression of toll like receptor-3 (TLR-3) in the bursa of Fabricius. In the fifth week, quail were challenged with 200 μL of E. coli O2 (1×10⁴ colony forming units (CFU)/mL).No clinical signs of Pb toxicity were observed. Morbidity/mortality subsequent to E. coli exposure was lowest in the high exposure group (27.8%) compared to low exposure (44.4%) and control (55.5%) groups. There was no difference in the T-cell-mediated PHA response, primary or secondary immune response or the innate response in Pb exposed groups; however, bursal TLR-3 increased (p<0.05) with higher Pb exposure.No evidence supported that subchronic Pb exposure was immunotoxic to quail at 5 or 50 ppm in drinking water. In contrast, our results provide evidence of a hormetic effect, with Pb exposed birds having lower morbidity and better survival than controls. Subchronic Pb exposure may be immunostimulatory rather than suppressive as predicted in earlier studies based on testing individual immune parameters.     

Toxicologic studies with japanese quail fed winter wheat grown on municipal sludge-amended soil

Winter wheat was grown on soil amended with 100 dry tons per acre (224 metric tons/ha) of municipal sewage sludge from Syracuse, New York. The grain contained 1.43 and 0.98 ppm (mg/kg) dry weight, respectively, of cadmium and nickel. This wheat was incorporated as 60% of a semi-synthetic diet and fed to male and female Japanese quail for two generations. Male quail from the F₁ generation fed sludge-grown wheat showed induction of hepatic microsomal enzymes,i.e., aminopyrene-N-demethylase and aniline hydroxylase, that indicated foreign compounds present in the wheat. Cadmium was significantly higher (p < 0.05) than controls in kidney, liver, and testes and nickel in the liver of the male quail (F₀) fed the sludge-grown grain. Cadmium in kidney and liver (but not in eggs) and nickel in liver was significantly higher (p < 0.05) than controls in the females fed the sludge-grown wheat. Birds from the F₁ generation showed no significantly different (p > 0.05) concentrations of cadmium in kidney, liver, or eggs between the two dietary treatments. There were no observable changes in the tissue ultrastructure of liver and kidney as examined by electron microscopy in any of the treatment groups.     

Reproductive Performance of Two Generations of Female Semidomesticated Mink Fed Diets Containing Organic Mercury Contaminated Freshwater Fish

Semidomesticated female mink (Mustela vison) were fed daily diets containing 0.1 ppm, 0.5 ppm, and 1.0 ppm of total mercury. Piscivorous and nonpiscivorous fish naturally contaminated with organic mercury were used to prepare the diets. Twenty-month-old females (G1 generation) that were exposed to the experimental diets for approximately 400 days in 1994 and 1995 and their 10-month-old female offspring (G2 generation) that were exposed to mercury for approximately 300 days in 1995, were all mated to 10-month-old males. Males were fed the diet containing 0.1 ppm mercury 60 days prior to the mating season. Diets containing 0.1 ppm and 0.5 ppm were not lethal to G1 and G2 females for an exposure period of up to 704 days. At the age of 11 months, mortalities occurred in 1994 for G1 females (30/50) and in 1995 for G2 females (6/7) fed the 1.0 ppm mercury diet after 90 days and 330 days of exposure, respectively. The length of the gestation periods and the number of kits born per female were not different among dietary groups for the two generations of females. The proportion of females giving birth was low for all groups, except for the G1 females fed the 0.1 ppm diet. There was an inverse relationship between whelping proportion and exposure group, but was not statistically significant. There was evidence that kits were exposed to mercury both in utero and/or during lactation as indicated by the presence of mercury in their livers. Mercury exposure did not influence the survival and growth of neonatal kits. Received: 11 November 1997/Accepted: 24 August 1998     

Effects of dietary exposure to fumonisins from Fusarium moniliforme culture material (M-1325) on the reproductive performance of female mink

Adult female mink (Mustela vison) were fed diets that contained Fusarium moniliforme culture material that provided low- or high-dose dietary concentrations of 86 or 200 ppm fumonisin B₁, 22 or 42 ppm fumonisin B₂, and 7 or 12 ppm fumonisin B₃, respectively, from approximately two weeks prior to breeding through gestation and lactation. Breeding performance of the females was not affected by consumption of the fumonisin diets. However, 58% of the mated females fed the high-dose diet (254 ppm total fumonisins) whelped compared to 100% of those fed the control and low-dose diets (115 ppm fumonisins). There was a statistically significant, dose-dependent decrease in kit (young mink) body weights at birth and a notable, but non-significant, decrease in litter size. The percentage of stillborn kits was directly proportional to the concentration of fumonisins in the dams' diets. Fumonisin concentrations in milk collected from those fed the high-dose diets were approximately 0.7% of the dietary fumonisin concentrations. Lactational exposure to fumonisins did not significantly decrease kit survival from birth through three weeks of age. Hepatic cell vacuolation was present in 25% of the control and 80% of the high-dose adults. No treatment-related gross or histologic lesions were observed in the kit mink. Numerous differences in hematologic and serum chemical parameters were noted between the control and fumonisin-exposed mink.     

Effect of mercury on erythrocyte glutathione reductase activity.In vivo andin vitro studies

Summary Glutathione reductase activity was determined in erythrocyte hemolysates of rats and quail fed various levels of methylmercury. Dietary mercury as high as 30 ppm had no effect on the activity of this enzyme. Micromolar concentrations of inorganic mercuric nitrate added to stroma-free hemolysatesin vitro resulted in a clear inhibition. To get the same decrease in activity with methylmercuryin vitro, however, it was necessary to add a hundred-fold higher level of Hg. Dietary selenium seemed to have some protective effect against the inhibitory effect of mercury addedin vitro.     

Pre- and Postnatal Exposure to 3,3′,4,4′-Tetrachlorobiphenyl: I. Effects on Breeding Ability and Sperm Fertilizing Ability in Male Mice

This study investigated the effects of 3,3′,4,4′-tetrachlorobiphenyl (TCB) on the reproductive performance and in vitro sperm fertilizing ability of male mice. C57BL/6J female mice (F-0) were fed diets containing 0, 3, or 30 ppm TCB for 2 weeks before pairing with nontreated males for a 10-day breeding period. Females were continued on their treatment diet throughout mating, gestation, and lactation. Male offspring (F-1) were given the same diet as their dams throughout this study. F-1 males at 7 and 17 weeks of age were mated to nontreated B₆D₂-F₁ female mice. Body weight, litter size, and survival indices of offspring sired by F-1 males were recorded. Sperm fertilizing ability of F-1 males at 9 and 19 weeks of age was examined in vitro using eggs collected from nontreated B₆D₂-F₁ females. Sperm motion analysis was performed at the same time. When 7- and 17-week-old F-1 males were mated to nontreated females, no differences in fecundity, litter size, sex ratio, or survival indices among any of the treatments were observed. However, sperm fertilizing ability of 30 ppm–treated F-1 males at 19 weeks of age was less than that of the control mice. Testes weights were greater in 3-week-old, 30 ppm–treated mice. Received: 16 August 1996/Accepted: 18 February 1997     

Assessment of the reproductive toxicology of bromochlorodifluoromethane (BCF, halon 1211) in the rat

The effect of bromochlorodifluoromethane (BCF) on reproduction in the rat has been investigated in two studies. Pregnant female rats were exposed by inhalation to 1000, 10,000, or 50,000 ppm BCF for six hours a day on days six to 15 of gestation (day of mating = day 0). Exposure to 50,000 ppm BCF caused a reduction in maternal weight gain over the exposure period but there was no evidence of either teratogenicity or embryo/fetotoxicity at any concentration. In a study designed to assess the potential effect of BCF during a complete reproductive cycle male and female rats were exposed to 5000 ppm or 25,000 ppm BCF for six hours a day for five days a week for 10 weeks (males) or three weeks (females) before mating. Exposure to BCF continued during mating and up to day 20 of gestation for half the females which were subsequently allowed to litter and the development of their offspring monitored. The remaining females were removed from exposure to BCF after mating and killed on day 20 of gestation for examination of their uterine contents. There were no effects on adult fertility, pup numbers, survival, or pup development. It was concluded that BCF had no reproductive toxicity potential in the rat.     

Assessment of the reproductive toxicology of bromochlorodifluoromethane (BCF, halon 1211) in the rat.

The effect of bromochlorodifluoromethane (BCF) on reproduction in the rat has been investigated in two studies. Pregnant female rats were exposed by inhalation to 1000, 10,000, or 50,000 ppm BCF for six hours a day on days six to 15 of gestation (day of mating = day 0). Exposure to 50,000 ppm BCF caused a reduction in maternal weight gain over the exposure period but there was no evidence of either teratogenicity or embryo/fetotoxicity at any concentration. In a study designed to assess the potential effect of BCF during a complete reproductive cycle male and female rats were exposed to 5000 ppm or 25,000 ppm BCF for six hours a day for five days a week for 10 weeks (males) or three weeks (females) before mating. Exposure to BCF continued during mating and up to day 20 of gestation for half the females which were subsequently allowed to litter and the development of their offspring monitored. The remaining females were removed from exposure to BCF after mating and killed on day 20 of gestation for examination of their uterine contents. There were no effects on adult fertility, pup numbers, survival, or pup development. It was concluded that BCF had no reproductive toxicity potential in the rat.     

Pre- and Postnatal Exposure to 3,3′,4,4′-Tetrachlorobiphenyl: II. Effects on the Reproductive Capacity and Fertilizing Ability of Eggs in Female Mice

This study investigated the effects of 3,3′,4,4′-tetrachlorobiphenyl (TCB) on the reproductive capacity of female mice. Female C57BL/6J mice (F-0) were fed diets containing 0, 3, or 30 ppm TCB for 2 weeks before pairing with nontreated C57BL/6J males for a 10-day breeding period. Females were continued on their treatment diet throughout mating, gestation, and lactation. Female offspring (F-1) were fed the same diet as their dams throughout the study. The reproductive capacity of F-1 females was examined by mating with non-treated B₆D₂-F₁ males. In addition, the fertilizing ability of eggs from F-1 females was examined in vitro by insemination with sperm from nontreated B₆D₂-F₁ males. Fecundity in F-0 females after mating was 80%, 71%, and 47% in the 0, 3, and 30 ppm treatment groups, respectively. Four-day and 21-day survival indices were lower for offspring of 30 ppm TCB-treated F-0 females than for offspring of the control females. Fecundity in F-1 females was the same among all treatment groups, however, all offspring born to 3- and 30-ppm TCB-treated F-1 females died before 4 days of age. Although the litter size at birth was not affected, the in vitro fertilizing ability of eggs in the 3- and 30-ppm treatment groups was lower than in the control group. This decrease in fertilizing ability was associated with an increase in degenerated eggs. F-0 females treated with 30 ppm TCB had enlarged livers during pregnancy and lactation. At 5 and 6 weeks, liver enlargement and thymus atrophy were apparent in F-1 females exposed to 30 ppm TCB. This study demonstrated impaired reproductive capacity and decreased egg fertilizing ability in TCB-treated female mice. Received: 15 May 1997/Accepted: 22 July 1997     

Effects of ammonium perchlorate on the reproductive performance and thyroid follicle histology of zebrafish

Adult zebrafish were reared up to eight weeks in control water or in water containing ammonium perchlorate (AP) at measured perchlorate concentrations of 18 (environmentally relevant, high) and 677 ppm. Groups of eight females were paired with four males on a weekly basis to assess AP effects on spawned egg volume, an index of reproductive performance. All treatments were applied to four to five spawning replicates. At 677 ppm, spawn volume was reduced within one week and became negligible after four weeks. At 18 ppm, spawn volume was unaffected even after eight weeks. Also, perchlorate at 18 ppm did not affect percentage egg fertilization. Fish were collected at the end of the exposures (677 ppm, four weeks; control and 18 ppm, eight weeks) for whole-body perchlorate content and thyroid histopathological analysis. Fish perchlorate levels were about one-hundredth of those of treatment water levels, indicating that waterborne perchlorate does not accumulate in whole fish. At 677 ppm for four weeks, perchlorate caused thyroid follicle cell (nuclear) hypertrophy and angiogenesis, whereas at 18 ppm for eight weeks, its effects were more pronounced and included hypertrophy, angiogenesis, hyperplasia, and colloid depletion. In conclusion, an eight-week exposure of adult zebrafish to 18 ppm perchlorate (high environmentally relevant concentrations) affected the histological condition of their thyroid follicles but not their reproductive performance. The effect of 677 ppm perchlorate on reproduction may be due to extrathyroidal toxicity. Further research is needed to determine if AP at lower environmentally relevant concentrations also affects the thyroid follicles of zebrafish.     

Subchronic toxicity and reproduction effects of tri-n-butyltin oxide in Japanese quail

A subchronic toxicity/reproduction study was performed in Japanese quails that were fed a diet containing 0, 24, 60, and 150 mg tri-n-butyltin oxide (TBTO) per kg basal diet for 6 weeks. Eggs produced during the 6 weeks of treatment were incubated and hatched, and chicks hatched from eggs collected in weeks 5 and 6 of exposure were reared for 2 weeks. In parent quail, neither diminished food consumption nor any overt toxic or histopathologic signs were observed following exposure to TBTO. A statistically significant decrease in hatchability and increase in percent of chicks found dead in the shell were observed following TBTO exposure at concentrations of 60 and 150 mg/kg food. However, no significant, adverse effects were recorded on total egg production, eggshell thickness and cracked eggs. Blood chemistry parameters of birds measured at the last day of TBTO treatment revealed a statistically significant decrease in serum aspartate aminotransferase (ASAT) enzyme activity among both sexes in all treatment groups. In addition, a statistically significant dose-related decrease in serum calcium level was observed in females only, while serum follicle stimulating hormone (FSH) levels were statistically significantly reduced in male birds in all treatment groups (approximately 50% of the controls).Moreover, a significant decrease in hepatic microsomal 7-ethoxyresorufin (EROD) activity was recorded in females fed 24 and 60 mg TBTO/kg diet and males fed 60 and 150 mg TBTO/kg diet, whereas pentoxyresorufin-o-deetylase (PROD) activity was only significantly decreased in males fed 150 mg TBTO/kg diet. No TBTO related effects were found on serum alanine aminotransferase (ALAT), serum total thyroxine (TT₄), luteinizing hormone (LH) or retinol levels in both sexes. In summary, TBTO affected mainly reproduction in Japanese quail at a dose where no overt toxicity is observed.     

Epoxiconazole causes changes in testicular histology and sperm production in the Japanese quail (Coturnix coturnix japonica)

The fungicide epoxiconazole (Epox), a triazole, belongs to the group of azole compounds that are extensively used as fungicides in various fruit crops. The frequent use of agricultural lands for wintering by migrating birds can be the source of their increased dietary intake of agricultural pesticides. We investigated whether exposure to Epox causes effects on avian fertility and reproduction, using the Japanese quail (Coturnix coturnix japonica) as a model species for the assessment of reproductive effects of pesticides in wild birds. Epox was administered to adult Japanese quail for three weeks at dietary levels of 10, 50, and 500 ppm, and possible effects on reproduction were investigated. Epox administration resulted in a significantly decreased number of spermatids in the 50- and 500-ppm dose groups. Histopathology showed a reduced number of testicular canaliculi with visible germ cells and a reduction in spermatid number. However, testis weight was not affected up to the highest dose level. No impact was observed on hormone levels, fertility, and reproductive outcome, as laying rate and percentage of fertile eggs were not altered. Likewise, treatment had no influence on the egg or chick parameters evaluated. A time- and dose-related transfer of Epox into the eggs was determined in all treatment groups. We conclude that dietary treatment of Japanese quail with 50 and 500 ppm of the triazole fungicide Epox resulted in a clear impact on the testis. The evaluation of the additional endpoints spermatid count and testicular histology have proven useful and are recommended for future studies on avian reproduction.     

Dietary selenium protection of methylmercury intoxication of Japanese quail

Summary Selenium, as sodium selenite, added at 5 ppm to purified diets of Japanese quail protected against methylmercury intoxication. Selenium fed simultaneously with methylmercury to quail for 9 weeks gave complete protection. However, feeding selenium with methylmercury for 4 weeks, followed by a diet containing only methylmercury, delayed the onset of methylmercury intoxication for 1–2 weeks as compared to quail not pretreated with selenium. On diets which contained 20 ppm of methylmercury but no selenium, over 90% mortality was observed for young quail within 2 weeks, and mature quail within 4 weeks. Methylmercury residues in liver, kidney, and brain are higher in male than female quail. High methylmercury content of these organs, or in produced eggs, does not indicate that birds will show evidence of methylmercury toxicosis.Approved by the Director of the New York State Agricultural Experiment Station for publication as Journal Paper No. 2012.     

Lead acetate induced reproductive and paternal mediated developmental toxicity in rats

Lead was administered orally to adult male rats at exposure level of 273 or 819 mg/L (0.05% or 0.15% lead acetate, respectively) for 45 days via drinking water. At the end of the exposure period, control and experimental males were mated with untreated females. Of the females mated with treated males, 73.3% in the 0.05% group and 53.33% in the 0.15% group showed copulatory plugs. Significant decrease in number of implantations and pre- and post-implantation loss was also observed in females mated with treated males. Significant decrease in the weight of the reproductive organs, reduction in epididymal sperm count, motile sperm and viable sperm were observed in lead-exposed rats indicating decreased sperm production and deteriorated sperm quality. Significant decrease in serum testosterone levels were also observed in treated rats indicating decreased steroidogenesis. The decreased serum testosterone levels and deteriorated sperm quality might be responsible for the suppressed reproduction in rats after exposure to lead.     

Residue uptake and depletion measurements of dietary oxadiazon in mammalian and avian species

The herbicide oxadiazon, 2-(tert-butyl)-4-(2,4-dichloro-5-isopropyloxy-phenyl)-²-1, 3, 4-oxadiazoline 5-one, was fed in a total diet at three dosage levels to groups of dairy cows (0.5, 2.5, and 25 ppm) and to groups of male and laying female quail (20, 80, and 160 ppm) for 28 days. Control animals received basal diet rations. No residue (<5 ppb) was detected in milk or cream after three days withdrawal from fortified feed. The maximum plateau reached in milk was 83.8 ppb at the highest dosage fed. Tissues of cows sacrificed on the 12th day of withdrawal showed no residue (<20 ppb). Birds were sacrificed at weekly intervals during acclimation, intake, and withdrawal periods. Eggs were collected at the same intervals. Tissues were free of oxadiazon residue (<40 ppb) within two to seven days of returning the quail to the basal ration, and the eggs were residue-free within seven to fourteen days. The maximum residue found in adipose tissue of quail and their fresh eggs was 3100 ppb and 5400 ppb, respectively at the greatly exaggerated dosage of 160 ppm in the diet. A comprehensive analytical methodology, including liquid-liquid partition, absorption chomatography and electron-capture gas chromatography will be discussed.Presented at the Third International Congress of Pesticide Chemistry (IUPAC), Helsinki, July 3–9, 1974.