Effects of the herbicide methabenzthiazuron on the physiology of wheat plants

Spring wheat (Triticum aestivum L. “Caribo”) was grown in vermiculite containing methabenzthiazuron (N-(benzothiazol-2yl)-NN′-dimethylurea) presowing. Effects of the herbicide on plant development and plant composition were analysed up to an age of 4 weeks. Inhibition of photosynthetic oxygen evolution represented the primary effect induced by the herbicidal treatment and led to a decreased concentration of soluble reducing sugars. Photosynthetic activity however recovered after 3 weeks and even increased above control values. Secondary effects following methabenzthiazuron treatment included a delayed chlorophyll breakdown, a decreased chlorophyll a/b ratio, enlarged chloroplasts, an increased concentration of soluble amino acids and of soluble protein, and an increased in vitro nitrate reductase activity. These responses are taken to indicate an increased photosynthetic and metabolic capacity in methabenzthiazuron treated wheat plants. Comparable results can be obtained with plants grown at low light intensities. It is concluded that the “physiological effects” observed in wheat plants after treatment with methabenzthiazuron are similar to a natural adaptation reaction to low light intensities. It is assumed that this adaptation reaction is caused by a low concentration of soluble reducing sugars. Experiments with plants growing at different light intensities indicated that effects due to herbicidal action were more pronounced at high light intensities. Measurements on daily fluctuations revealed a peak around noon for the sugar content and the nitrate reductase activities measured in vivo as well as in vitro. In vivo nitrate reductase activity in plants treated with 5 parts/million methabenzthiazuron was very low, presumably because of lack of sugars for the production of NADH. The protein concentration was increasing and the amino acids were decreasing during the day in herbicide treated plants, possibly indicating increased protein synthesis in the light in plants treated with methabenzthiazuron.     

Involvement of heme synthesis in the growth stimulation of maize seedlings by 5‐aminolevulinic acid

The growth of maize seedlings was stimulated by shoot‐applied 5‐aminolevulinic acid 2 days after treatment at 90 and 120 μmol L^?1. The effects of 5‐aminolevulinic acid on the activity of nitrate reductase (EC 1.6.6.2) and nitrite reductase (EC 1.7.7.1) and the chlorophyll, ammonium, heme and total free amino acid content were investigated by using maize seedlings to clarify the involvement of nitrogen metabolism and heme synthesis in the growth stimulation. 5‐Aminolevulinic acid increased the level of nitrate reductase activity at 90 μmol L^?1 and the ammonium and heme content at 90 and 120 μmol L^?1 2 days after treatment. The total amino acid content increased by 90 and 150 μmol L^?1 5‐aminolevulinic acid 2 and 3 days after treatment, respectively. However, no significant change was observed in the activity of nitrite reductase or the chlorophyll content after the 5‐aminolevulinic acid treatment. These results suggest that the enhancement of nitrogen metabolism by nitrate reductase activation is involved in the 5‐aminolevulinic acid‐induced stimulation of maize seedling growth. The activation of nitrate reductase might be related to an increase in the heme content following the 5‐aminolevulinic acid treatment.     

Effects of ametryn [2-(ethylamino)-4-(isopropylamino)-6-(methylthio)-s-triazine] on nitrate reductase activity and nitrite content of wheat (Triticum aestivum L.)

The objectives of this study were to show that: (a) a herbicide, such as ametryn, which interferes with the photosynthetic electron transport system, causes nitrite to accumulate in illuminated leaves and (b) that nitrite is toxic and contributes to the herbicidal damage and death of the plant. Tests were conducted on wheat seedlings grown on 5 mM nitrate, 5 mM ammonia, and zero nitrogen. Ametryn treatment decreased in vivo and in vitro nitrate reductase activity (NRA) within a 26-hr period. In vivo NRA decreased more rapidly than in vitro NRA. Compared with control tissue, only 3% in vivo NRA remained at the end of 26 hr. The in vivo assay conducted in light confirmed the inhibition of photosynthetic electron flow by ametryn within the leaf tissue. Nitrate-grown, ametryn-treated plants accumulated nitrite and, after 10 days were the only plants that were completely desiccated and dead. Ammonia- and zero-nitrogen, ametryn-treated plants did not accumulate nitrite, were only partially chlorotic after the 10-day period, and were still living. Low levels of NO_(X) (NO₂ and/or NO) emissions were demonstrated by nitrate-grown ametryn-treated plants.     

Induction of glutathione S-transferase by phenobarbital and pesticides in various house fly strains and its effect on toxicity

The effect of phenobarbital and certain pesticides on glutathione S-transferase activity was investigated. The maximum amount of enzyme induction occurred 96 hr after phenobarbital treatment. Chlorinated hydrocarbons were more effective inducers than the other pesticides evaluated. Phenobarbital treatment did not alter the apparent K_m value but altered the $\text{V}{}_{\mathrm{<mtext>max</mtext>}}$ value of glutathione S-transferase to 3,4-dichloronitrobenzene. The amount of reduced glutathione was not increased by phenobarbital treatment. Pretreatment of house flies with phenobarbital provides some protection against methyl parathion, methyl paraoxon, azinphosmethyl, and methidathion toxicity.     

Cytochrome P-450 content and aldrin epoxidation to dieldrin in isolated rat hepatocytes

A rat hepatocyte suspension effectively epoxidized aldrin to dieldrin with a V_max of 7.19 mol/mol P-450/min and a K_m of 9.27 μM. Viability and metabolic activity were stable for 6 hr after isolation when cells were maintained at room temperature (20°C) with the gentle introduction of $\text{O}{}_2\text{CO}{}_2$ onto the surface of the suspension. The cytochrome P-450 content of the suspension was 303 pmol/10⁶ cells. Primary maintenance culture of the cells also epoxidized aldrin. During culture for 3 days, metabolic activity decreased slowly day by day. Metabolic activity of microsomal fraction from rat liver was also examined. Microsomes epoxidized aldrin with a V_max of 5.11 mol/mol P-450/min and a K_m of 1.64 μM. Significant loss of some subspecies of cytochrome P-450 during fractionation of liver homogenate was indicated.     

The metabolism of p,p′-DDT by the feral pigeon (Columba livia)

Male feral pigeons were dosed with ring-labeled $\text{[}{}^{14}\text{C}\text{]p,p′-}\text{DDT}$ and the tissues and droppings analyzed for total ¹⁴C, extractable ¹⁴C, and metabolites. Only 16% of an intraperitoneal dose of 1.5–2.2 mg kg^?1 was voided in the droppings over 28 days; the rate of loss reached a maximum on the 14th day and then fell quickly away. The rate of removal of ¹⁴C in droppings was low in comparison to that found in the rat and the Japanese quail. When pigeons were dosed with 32–38 mg kg^?1 DDT per bird, and killed after 77 days, 5.4% of the dose was eliminated in droppings and 87% was recovered in the body. The tissues and droppings from this experiment were analyzed for DDT and its metabolites. Of the ¹⁴C remaining in tissues 88% was accounted for as the apolar compounds DDE, DDT, and DDD. Approximately half of the ¹⁴C in droppings was present as DDE, DDT, and DDD, whereas 27–35% was apparently in conjugated form, extractable from aqueous solutions by ethyl acetate after prolonged acid hydrolysis. Two polar metabolites were isolated from the acid-released material. One was p,p′-DDA; the other was extractable from aqueous solution at pH 8 and was tentatively identified as a monohydroxy derivative of p,p′-DDT. DDE accounted for 93% of the ¹⁴C present as metabolites in tissues and droppings, clearly indicating the importance of this intermediate in this study. The metabolism of DDT in the feral pigeon is discussed in relation to its metabolism by other species.     

Components of the electron transport system in the microsomal mixed-function oxidase system in wild birds

Notable differences were found among six species of wild-caught birds in the levels of cytochrome P-450, cytochrome b₅, NADPH-cytochrome c reductase, and NADH-cytochrome c reductase. Ethyl isocyanide difference spectra showed significant variations among the species in peak height and in the ratios of the $\text{430}\text{455}\text{-}\text{nm}$ peaks. Substantial aldrin epoxidase activity was found in all species, and the amounts of dieldrin produced compared favorably with pigeon and rat liver microsomes. Higher content of cytochrome P-450 was not always accompanied by a similar rise in specific catalytic activity. Thus, no correlation could be established between these two parameters. Aldrin epoxidase activity with NADH as the sole electron donor was 25–49% as effective as with the NADPH-generating system. Addition of both NADH and NADPH-generating systems to the incubation mixture produced a synergistic effect with liver microsomes of two species but not with two other species. DDE and polychlorinated biphenyls residues were found in the heart tissue of all species examined, and this might indicate a possible inductive effect on the microsomal mixed-function oxidase system by environmental contaminants.     

Mitochondrial changes during storage of untreated or CIPC-treated potatoes

During storage of potato tubers (Solanum tuberosum L., var. Bintje), important changes appear which affect respiratory control, $\text{ADP}\text{O}$, intensity of O₂ consumption in the presence of different substrates, and NAD⁺ dependence, In mitochondria extracted under strictly similar conditions, from patato tubers stored at 4°C, the respiratory control (RC) maintains a value near 4 for 4 to 5 months. It then declines progressively to low values. At 20°C, a stable RC of 4 can be observed for several months, which then decreases at the end of dormancy. Then, the RC increases sharply; at this stage, $\text{ADP}\text{O}$ are abnormally low, and, some time later, NAD⁺ dependence disappears. Mitochondria treated with 250 μM chlorpropham show a 50% inhibition of the electron transfer with exogenous NADH as substrate. After tuber treatment with 1% chlorpropham, sprouting is inhibited for several months. The activities of mitochondria extracted from such tubers remain unaffected by the treatment. The use of this phenylcarbamate for potato tuber treatment permits obtaining functional mitochondria from tubers after a slightly longer period of storage.     

Identification of products arising from the metabolism of cis- and trans-chlordane in rat liver microsomes in vitro: Outline of a possible metabolic pathway

The metabolism of pure cis- and trans-chlordane was studied in vitro. Microsomal preparations from the livers of male rats induced with cis- or trans-chlordane in feed for 10 days were used to metabolize the pure compound corresponding to the inducer. Subsequent extraction, column fractionation, and combined gas chromatography-mass spectroscopy resulted in the characterization of four compounds not previously reported from an in vitro system. In addition to the substrate, trans-chlordane extracts contained species with the following molecular weights and empirical formulas: $\text{m}\text{e}$ 370, C₁₀H₅Cl₇, heptachlor; $\text{m}\text{e}$ 352, C₁₀H₆OCl₆, a hydroxylated chlordene; and $\text{m}\text{e}$ 422, C₁₀H₆OCl₈, a hydroxylated chlordane. Dichlorochlordene, oxychlordane, and 1-chloro-2-hydroxy-dihydrochlordene were also present. With the exception of the hydroxychlordane, cis-chlordane extracts contained all of the metabolites found in the trans incubates. Additionally, a fully saturated compound, $\text{m}\text{e}$ 372, C₁₀H₇Cl₇, a dihydroheptachlor, was present. The 1,2-trans-dihydrodiol of heptachlor found in previous in vitro incubates of cis-chlordane was not present in this extract. This information has been incorporated into a proposed route for the biotransformation of the chlordanes that offers an explanation for the observed differences in the metabolism of cis and trans isomers. The pathway is based on the reductive dechlorination of the chlordanes through dihydroheptachlor to dihydrochlordene. Parallel pathways of hydroxylation, desaturation, and epoxide formation arise at each of these species and at chlordane itself.     

Mode of action of bentazon: Effect on photosynthesis

Bentazon, 3-isopropyl-2,1,3-benzothiadiazin-4-one-2,2-dioxide is effective for weed control in flooded rice fields not only as a foliar treatment but also as a flooded-water or paddy soil treatment. Generally the herbicidal effect develops slowly only after translocation of the herbicide has occurred, but when the weeds contacted directly with relatively high concentrations of the herbicide, the effects appear rather rapid.The slow herbicidal effect appears to be an important mode of action of bentazon applied practically on weeds under flooded rice field conditions. The slow effect may be caused by inhibition of photosynthesis as supported by the following experimental results: a) Bentazon inhibited the Hill reaction in isolated chloroplasts; b) bentazon rapidly inhibited photosynthetic CO₂ fixation in susceptible Cyperus serotinus and other plants; c) the herbicidal effects appeared much slower when bentazon was applied as a flooded-water treatment; d) bentazon injury was prevented by endogenous or exogenously supplied carbohydrates.     

Mechanism of inhibition reaction of acetylcholinesterase by phenyl N-methylcarbamates: Separation of hydrophobic,electronic, hydrogen bonding and proximity effects of aromatic substituents

The association equilibrium constant, $\text{1}\text{K}{}_{\mathrm{d}}$, and the carbamylation constant, k₂, of 53 o-, m-, and p-substituted phenyl N-methylcarbamates with bovine erythrocyte acetylcholinesterase were determined. The $\text{1}\text{K}{}_{\mathrm{d}}$ value varied 1000-fold, whereas the k₂ value did not depend upon the nature and position of substituents. The variation in $\text{log}\text{(}\text{1}\text{K}{}_{\mathrm{d}}\text{)}$ was analyzed using free energy related substituent parameters and regression analyses. The effect of substituents at o-, m-, and p-positions was nicely separated into hydrophobic, electronic, hydrogen bonding, and proximity (steric and field electronic for o-substituents) factors. The physicochemical significance of these factors was established by comparison with those for model organic reactivities. The mechanism of the whole reaction process was elucidated in terms of physical organic chemistry.     

Importance of pH in assessing the potential for nitrosocarbamate formation in the stomach

Formation of the nitroso derivatives of ¹⁴C-labeled carbaryl, carbofuran, and p-chlorophenyl methylcarbamate (PCMC), and their subsequent stabilities were investigated using low levels of carbamate (0.5 μmol) and aqueous HCl concentrations encompassing those considered maximum for the gastric contents of humans (pH 1–2) and of rats (pH 3–4). Reacting the carbamates with excess sodium nitrite for 10 min at 37°C in a pH 1.0 HCl solution gave nitrosocarbamate yields of 42 to 64%, while only trace amounts were formed at pH 2 and above. The nitrosocarbamates were most stable at pH 3–5 with half-lives ranging from 114 to 470 min. Stability of all three nitroso derivatives was considerably less at pH 1.5 ($\text{t}\text{1}\text{2}$, 25–34 min), but at pH 7 the stability varied: nitroso-PCMC $\text{t}\text{1}\text{2}$, 6 min; nitrosocarbofuran $\text{t}\text{1}\text{2}$, 70 min; nitrosocarbaryl $\text{t}\text{1}\text{2}$, 139 min. Denitrosation to the parent carbamate was the predominant degradation pathway at pH 1.5, but at pH 3–7 degradation was primarily by hydrolysis of the carbamate ester linkage. Each of the nitrosocarbamates was directly mutagenic in the S. typhimurium assay system. Since the data show that nitrosation of residue levels of carbamate pesticides occurs readily at pH 1 but not at pH 2 and above, it is critical that gastric contents of any animal model used for assessing nitrosocarbamate formation have a pH approaching 1 as may occur in the human stomach.     

Interaction of insecticides with the Ca2+-pump activity of sarcoplasmic reticulum

The organophosphorus insecticides, parathion and azinphos (10^?5-10^?4M), significantly stimulate the Ca²⁺-pump activity of sarcoplasmic reticulum, while malathion has a limited effect. The rates of Ca²⁺ translocation and ATP hydrolysis are both stimulated and, apparently, the $\text{Ca}{}^{\mathrm{2+}}\text{ATP}$ ratio is improved. Parathion and azinphos maximally increase this ratio by 26 and 14%, respectively. The organochlorine compounds, DDT and aldrin, also stimulate the Ca²⁺ pump, and lindane has a reduced effect. These effects are smaller than those observed for parathion and azinphos. The order of effectiveness is similar to the toxicity of the compounds to mammals and can be described as follows: parathion > azinphos > DDT ≈ aldrin > malathion ≈ lindane.     

干旱和复水对两种葡萄砧木叶片光合和叶绿素荧光特性的影响

采用盆栽称重控水法,干旱胁迫处理1103P和101-14M两种砧木,处理21 d后复水,分别测定干旱处理0、7、14、21 d及复水第7、14 天,葡萄砧木叶片光合和叶绿素荧光参数。结果显示：干旱胁迫后,1103P和101-14M的净光合速率(Pn)均逐渐降低,101-14M的Pn降幅大于1103P,短时间干旱胁迫引起两种砧木Pn降低的主要因素是气孔限制,而长时间干旱胁迫后Pn降低主要是非气孔限制。随着干旱胁迫的持续,1103P和101-14M的初始荧光产量(Fo)呈增加趋势,但101-14M的增幅大于1103P,说明干旱胁迫后101-14M的光反应中心受损害程度大于1103P;复水后1103P和101-14M两种砧木的Pn值逐渐增加,复水第7天,二者分别为对照的83.20%和66.31%,复水第14天,分别为对照的107.30%和88.43%; 复水后1103P和101-14M两种砧木的Fo值呈现逐渐降低趋势,复水后第7天,1103P和101-14M 的Fo值为对照的102.95%和109.60%,复水后第14天,1103P和101-14M Fo为对照的101.56%和101.81%,说明复水后1103P和101-14M两种砧木受损的光合反应中心得到了修复,光合速率也逐渐恢复,1103P复水后恢复生长的能力高于101-14M。     

In vivo biochemical changes in liver and gill of Clarias batrachus during cypermethrin exposure and following cessation of exposure

The sublethal effect of a synthetic pyrethroid, cypermethrin on total protein, amino acids, ammonia, glycogen, and enzymes like aminotransaminases (AIAT, AAT), glutamate dehydrogenase, and glycogen phosphorylases (a and ab) was studied in physiological important tissues viz; liver and gill tissues of freshwater teleost air breathing fish, Clarias batrachus. The study was conducted during exposure of 1/3 (33%) of LC₅₀ concentration and followed by cessation of exposure. Thirty-six fish were exposed to 0.07 mg/L cypermethrin for 10 days. After 10 days, 18 fish were released to freshwater and kept in the same for 10 days (recovery group). Thirty-six fish were kept in freshwater as control batch. Protein content in liver tissues decreased at the end of 1st and 5th day followed by slight increase at the end of 10th day. Gill tissue showed statistical significant decrease (P < 0.001) in protein content during exposure period of 10 days. Recovery in protein content was observed to a large extent in both the tissues. Total free amino acids were increased in liver and gill tissues throughout the treatment period, recovery response was seen after cessation of exposure. Ammonia level was decrease in both the tissues throughout the exposure period except in liver tissue at the end of 1st day of exposure. Recovery response was exhibited by both the tissues. A decreased in glycogen content of liver tissue was observed during exposure period, gill tissue also showed decrease in glycogen at the end of 1st and 5th day followed by increase at the end of 10th day of exposure period. When the fish were transferred to freshwater, recovery in glycogen content was noted. The activity level of alanine, aspartate aminotransaminase, glutamate dehydrogenase, and phosphorylases (a, ab) was increased in both the tissues, followed by recovery response after released of fish into freshwater. The present study showed that cypermethrin caused alterations in certain biochemical mechanisms of C. batrachus. This fish indicated recovery response when transferred to cypermethrin free water.     

The degradation of dinitramine (N3,N3-diethyl 2,4-dinitro-6-trifluoromethyl-m-phenylenediamine) in soil

Radioactive dinitramine (1) was incorporated at $\text{1}\text{2}$ Ib/acre (0.6 ppm) in Anaheim silty loam soil and its degradation studied over an 8-month period. For both specifically—¹⁴CF₃ and -Ring-UL-[¹⁴C] labeled (1), only ca. 20% of the radioactivity was lost from the incorporated zone. Mehanol- or acetonitrile-extractable radioactivity decreased rapidly over the initial 60 days reaching 20% after 244 days. Two compounds were isolated and characterized as (1), 0.05 ppm, and 6-amino-1-ethyl-2-methyl-7-nitro-5-trifluoromethylbenzimidazole (2), 0.06 ppm. Two other compounds were tentatively identified by TLC as monodealkylated dinitramine (3), 0.01 ppm, and 6-amino-2-methyl-7-nitro-5-trifluoromethylbenzimidazole (4), 0.01 ppm, Sodium hydroxide (10%) and anionic surfactant (10%) were effective in removing up to 50% of the residual bound radioactivity (i.e., nonacetonitrile extractable), while dimethylamine (25%) released 26%; extraction by acid was less effective.     

The purification and characterization of esterases from insecticide-resistant and susceptible house flies

Four major esterases in one susceptible (CSMA) and two resistant (Hirokawa, E₁) house fly strains were separated by chromatofocusing. Of the four esterases, those with pI's of 5.1 and 5.3 accounted for 90% of the p-nitrophenyl butyrate hydrolyzing activity in the three house fly strains. They also accounted for 70% (Hirokawa, E₁) and 40% (CSMA) of the paraoxon-hydrolyzing activity as well as 87% (Hirokawa), 39% (E₁) and 66% (CSMA) of the malathion-hydrolyzing activity in microsomes as measured by esterase-antibody interaction. In the Hirokawa strain, the pI 5.1 esterase was the predominant esterase and was more active than that of the the CSMA strain. Different substrate specificities and a different K_m toward acetylthiocholine, as well as different rates of malathion and paraoxon hydrolysis between the Hirokawa and CSMA strains, suggest a qualitative difference in the pI 5.1 esterase. For the pI 5.1 esterase from the E₁ strain, a different substrate specificity, a different K_m for p-nitrophenyl butyrate, a different sensitivity to inhibitors, and a different rate of paraoxon hydrolysis suggest that it is a modified esterase. This esterase is not a phosphorotriester hydrolase, nor does it lack nonspecific esterase activity. It is a modified esterase which has a different substrate specificity when compared to the esterases from the other strains. The molecular weight of the esterases studied was approximately 220,000, with pH optima of about 7.0.The ratio of malathion α-monoacid to β-monoacid formation was about 9.0 for the pI 5.1 and 5.3 esterases and 1.5 for the pI 4.8 and 5.6 esterases. The existence of a higher $\text{α}\text{β}$ ratio for the pI 5.1 and 5.3 esterases and their significant rate of malathion hydrolysis in the Hirokawa strain indicate that an increase in the $\text{α}\text{β}$ ratio in house flies reported was due to the increase in the pI 5.1 esterase in the resistant strain.     

Blood meal and cytochrome P-450 monooxygenases in the northern house mosquito,Culex pipiens

Conditions for the measurement of aldrin epoxidation by microsomes prepared from abdominal tissues (fat body + integument) of adult female Culex pipiens were characterized. The enzyme activity had a pH optimum of 7.2 and an apparent K_m of 3.4 μM. Aldrin epoxidation and NADPH-cytochrome c reductase had similar patterns of inhibition by a rabbit antiserum to house fly NADPH-cytochrome P-450 reductase, thus implicating cytochrome P-450 monooxygenase(s) in the epoxidation of aldrin. Low (71 pmol/mg protein) levels of cytochrome P-450 were detected in abdominal tissue microsomes. In non-blood-fed insects, aldrin epoxidation and NADPH-cytochrome c reductase activities did not change between Day 1 and Day 12 after adult emergence, except for a small peak on Day 2. In insects fed a blood meal on Day 6 after emergence both activities increased (two- to threefold) to a plateau maintained between 2 and 4 days after the blood meal. Aldrin epoxidation and NADPH-cytochrome c reductase activities decreased to normal values between 4 and 6 days after the blood meal.     

Spectral and inhibitory interactions of methylenedioxyphenyl compounds with southern armyworm (Spodoptera eridania) midgut microsomes

Characteristics of the Type III optical difference spectra of 13 methylenedioxyphenyl compounds in NADPH-fortified armyworm midgut microsomes varied with the nature of the substituents in the aromatic ring. Compounds with electron-donating substituents yielded spectra with large $\text{427}\text{458}\text{nm}$ peak ratios, whereas those with electron-withdrawing groups exhibited low $\text{427}\text{458}\text{nm}$ peak ratios. Small amounts of carbon monoxide were generated during incubation of the 4,5-dihalo derivatives with midgut microsomes, and cis- and trans-methylenedioxycyclohexanes exhibited spectra with a major Soret peak at about 430 nm and a very weak absorbance maximum at about 480 nm. Formation of the Type III spectral complex occurred very rapidly and was associated with a marked decrease (up to 72%) in cytochrome P-450 levels as measured by carbon monoxide binding. Although a 24% reduction of cytochrome P-450 was observed in the absence of any measureable 458-nm spectral complex a linear relationship existed between further decreases in the cytochrome and the increase in Type III complex formation (458 nm). Inhibitory potencies of the compounds towards aldrin epoxidase and benzopyrene hydroxylase activities were not clearly correlated with either spectral complex formation or decrease in cytochrome P-450 and it is apparent that different factors are involved in the inhibition of different monooxygenase reactions.     

Metabolism of lindane in house flies: Metabolic desaturation,dehydrogenation and dehydrochlorination,and conjugation with glutathione

In lindane-treated house flies, a cis-dehydrogenated metabolite, $\text{(}\text{36}\text{45}\text{)-}\text{hexachlorocyclohexene}$, was identified by gas-liquid chromatography and mass spectrometry. The in vitro metabolism study showed that in the presence of NADPH the microsomal fraction of house flies converted lindane to three hexane-soluble metabolites. This conversion was inhibited by piperonyl butoxide, SKF-525A, and carbon monoxide. These metabolites were identified as $\text{(}\text{36}\text{45}\text{)-}\text{hexachlorocyclohexene}$, $\text{(}\text{36}\text{45}\text{)- and (}\text{346}\text{5}\text{)-pentachlorocyclohexene}$ (PCCHE) by gas-liquid chromatography. They, as well as lindane, were excellent substrates for the reaction with the postmicrosomal fraction in the presence of glutathione. While the reaction with lindane-d₆ showed a significant deuterium isotope effect (6.82), that of $\text{(}\text{36}\text{45}\text{)-}\text{PCCHE-d}{}_5$ did not (1.18). Enzymatic conjugation with glutathione probably occurs at the stage of PCCHE.