Resistance of earthworms Lumbricus terrestris and Allolobophora turgida to the fungicide captan 50 W.P

Two species of earthworms were kept in Muck, Chicot and Ste-Sophie soils treated with captan 50 W.P. solutions of 700, 1 400 and 2 800 ppm. After a 42-day treatment period, L. terrestris had a 95% survival and A. turgida 100% survival. Using a gas chromatograph method, no captan was detected in tissue extracts of exposed earthworms. Based on the survival rate and the undetected presence of captan in earthworm tissues, we concluded that soil residues of this product are unlikely to be harmful to the animal's ecology.     

Captan alters transcription in Escherichia coli permeabilized by toluene

RNA synthesis was measured in toluenized E. coli by the incorporation of radiolabeled precursor into either acid precipitable or phenol extracted RNA. Exposure to captan (100 microM) caused a 2.6 fold increase in the apparent rate of RNA synthesis. When captan was tested for its effect on the initiation of RNA synthesis, using either rifampicin-treated cells or by measuring the incorporation of gamma [32P]ATP or gamma [32P]GTP, no change was observed in the number of RNA chains being initiated. Thus, captan does not exert its influence at the level of initiation of nascent chains. However, captan did have an effect on chain growth. From calculations of the incorporation of precursors molecules, RNAs isolated from treated cells were measured to be an average of 2.7 times longer than those from untreated cells. RNA chain lengths were also analyzed by polyacrylamide gel electrophoresis. By this latter technique it was also shown that cells exposed to captan synthesized RNAs that were longer than those of untreated cells. Alterations in the degradation of RNA molecules do not account for the captan mediated response in RNA synthesis.     

Interaction of captan with mammalian microtubules

Using turbidometry, electron microscopy and immunofluorescent microscopy experiments we studied the effect of captan, a widely used pesticide on mammalian microtubules and microfilaments. Turbidometry at 350 nm showed a dose-dependent inhibition of tubulin assembly incubated with captan. The pesticide, given at equimolar concentration with tubulin (30 microM), caused the total inhibition of microtubule formation, while at lower concentrations (5-20 microM) the inhibition of tubulin polymerization was less extensive. At the same concentration range (5-30 microM), captan also promoted the disassembly of performed microtubules. The results of the in vitro effects of captan with microtubules were confirmed in parallel by electron microscopic studies. In vivo, captan caused also depolymerization of microtubules in cultured mouse fibroblasts as shown by indirect immunofluorescent staining of tubulin. The extent of microtubules disassembly was concentration- and time-dependent. While incubation of the cells with 10 microM captan for 3 h disturbs totally the microtubular structures, incubation with 5 microM captan needs 12 h for the same effect. Recovery of microtubules was observed, when preincubated cells were extensively washed. No interaction of this drug with equimolar concentration of G- or F-actin could be observed in vitro, as shown by polymerization experiments. In line with this, the fluorescent actin pattern in mouse fibroblasts incubated with 10 mM captan for up to 12 h did not seem to be altered. From these results it is concluded that captan interacts in equimolar concentrations with tubulin affecting the assembly and disassembly of microtubules in vitro and in cultures of mammalian cells.     

Cytogenetic and dominant lethal studies on captan.

Possible mutagenic activity of captan was investigated by in vitro and in vivo cytogenetic studies and by the dominant lethal study in mice. In vitro cytogenetic study with cultured human diploid cells revealed a significant increase in the frequency of cells showing stickiness and a severe mitotic inhibition at concentrations of 3.0 and 4.0 microgram of captan per ml. although no chromosomal aberrations were observed. In in vivo cytogenetic study, no chromosomal aberrations were induced in the bone marrow cells of rats treated orally with captan at a single dose of 500, 1000 or 2000 mg/kg or at five consecutive doses of 200, 400 or 800 mg/kg/day. Dominant lethal study also failed to show any mutation induction after treatment of male mice with daily oral dose of 200 or 600 mg of captan per kg bw for five days.     

小熊猫繁殖周期血清雌二醇和孕酮含量变化

为研究小熊猫繁殖周期血清雌二醇、孕酮含量变化规律,采用化学发光免疫分析法连续16 次测定了2只成体雌性小熊猫血清雌二醇和孕酮含量变化,历经发情间期、发情期和两次妊娠期;连续9次测定了7只小熊猫妊娠期的孕酮含量变化。结果:(1)发情间期,小熊猫血清雌二醇的水平一直维持在低水平(基础水平),进入发情前期,血清雌二醇水平明显升高,在发情期一直维持高水平,配种后迅速降至基础水平; (2)小熊猫血清孕酮含量在发情间期和发情期均维持在较低水平,直至发情期过后才出现升高,在妊娠期一直维持高水平,峰值出现在5 月;(3)发情的小熊猫不论妊娠与否,在妊娠期内血清孕酮含量均维持在高水平。研究表明:小熊猫血清雌二醇、孕酮含量变化能直接反映其繁殖规律,雌二醇对启动雌性小熊猫季节性繁殖起重要作用;在妊娠期内小熊猫血清孕酮含量升高不能作为判断小熊猫妊娠的标准;雌性小熊猫在妊娠期有假孕现象。     

Binding of captan to DNA polymerase I from Escherichia coli and the concomitant effect on 5'----3' exonuclease activity

Captan (N-[(trichloromethyl)thio]-4-cyclohexene-1,2-dicarboximide) was shown to bind to DNA polymerase I from Escherichia coli. The ratio of [14C] captan bound to DNA pol I was 1:1 as measured by filter binding studies and sucrose gradient analysis. Preincubation of enzyme with polynucleotide prevented the binding of captan, but preincubation of enzyme with dGTP did not. Conversely, when the enzyme was preincubated with captan, neither polynucleotide nor dGTP binding was blocked. The modification of the enzyme by captan was described by an irreversible second-order rate process with a rate of 68 +/- 0.7 M-1 s-1. The interaction of captan with DNA pol I altered each of the three catalytic functions. The 3'----5' exonuclease and polymerase activities were inhibited, and the 5'----3' exonuclease activity was enhanced. In order to study the 5'----3' exonuclease activity more closely, [3H]hpBR322 (DNA-[3H]RNA hybrid) was prepared from pBR322 plasmid DNA and used as a specific substrate for 5'----3' exonuclease activity. When either DNA pol I or polynucleotide was preincubated with 100 microM captan, 5'----3' exonuclease activity exhibited a doubling of reaction rate as compared to the untreated sample. When 100 microM captan was added to the reaction in progress, 5'----3' exonuclease activity was enhanced to 150% of the control value. Collectively, these data support the hypothesis that captan acts on DNA pol I by irreversibly binding in the template-primer binding site associated with polymerase and 3'----5' exonuclease activities. It is also shown that the chemical reaction between DNA pol I and a single captan molecule proceeds through a Michaelis complex.(ABSTRACT TRUNCATED AT 250 WORDS)     

Mechanism of inhibition of Escherichia coli RNA polymerase by captan.

Captan (N-trichloromethylthiocyclohex-4-ene-1,2-dicarboximide) was shown to inhibit RNA synthesis in vitro catalysed by Escherichia coli RNA polymerase. Incorporation of [gamma-32P]ATP and [gamma-32P]GTP was inhibited by captan to the same extent as overall RNA synthesis. The ratio of [3H]UTP incorporation to that of [gamma-32P]ATP or of [gamma-32P]GTP in control and captan-treated samples indicated that initiation was inhibited, but the length of RNA chains being synthesized was not altered by captan treatment. Limited-substrate assays in which re-initiation of RNA chains did not occur also showed that captan had no effect on the elongation reaction. Studies which measured the interaction of RNA polymerase with template DNA revealed that the binding of enzyme to DNA was inhibited by captan. Glycerol-gradient sedimentation of the captan-treated RNA polymerase indicated that the inhibition of the enzyme was irreversible and did not result in dissociation of its subunits. These data are consistent with a mechanism in which RNA polymerase activity was irreversibly altered by captan, resulting in an inability of the enzyme to bind to the template. This interaction was probably at the DNA-binding site on the polymerase and did not involve reaction of captan with the DNA template.     

Integrated management of fusarial wilt of tomato with implementation of soil solarisation

Fusarial wilt of tomato (Lycopersicon esculentum Mill.) is a very common and severe disease occurring in most of the vegetable fields in West Bengal, India. Potenciation and formulation of different fungicidal chemicals and phytoextracts were evaluated against the growth of the pathogen wherein carbendazim (bavistin) and leaf extracts of Azadirachta indica (neem) were recorded to be most effective. Combined treatment with 4 ml neem leaf extract and 1 ml captan (0.01%) or with 4 ml garlic bulb extract and 1 ml captan (0.01%) exhibited 100% growth inhibition of the pathogen. Integrated control of the pathogen with phytoextracts, fungicide and biocontrol agents was carried out. Among the treatments, a combination with extracts of neem, captan (0.01%) and metabolites of Trichoderma harzianum was proved to be superior over the other. Field experiment with three fungicides at 0.5% concentration was carried out in randomised block design where application of bavistin showed up to 62.27% reduction of wilt infection in tomato plants. Soil solarisation of tomato field showed 62.50 and 66.69% reduction of infection during the trial years. However, integration of soil solarisation with the applications of T. harzianum, captan (0.01%) and neem resulted in 100% reduction of infection and thus it was recorded as the most effective treatment in reducing the incidence of the disease.     

Optical biosensor for simultaneous detection of captan and organophosphorus compounds

The optical biosensor consisting of GST and acetylcholinesterase (AChE)-immobilized gel film was developed to detect captan and organophosphorus compounds simultaneously in contaminated water. The sensing scheme was based on the measurement of decrease of products formation (s-(2,4-dinitrobenzene) glutathione and alpha-naphthol by GST and AChE, respectively) due to the inhibition by captan and organophosphorus compounds. The absorbance of s-(2,4-dinitrobenzene) glutathione and alpha-naphthol was detected at 400 and 500 nm, respectively, by a proposed optical biosensor system. It was observed that AChE was inhibited by both captan and organophosphorus compounds, and GST was inhibited only by captan. The simultaneous detection and quantification of captan and organophosphorus compounds could be successfully achieved by the proposed sensor system. The proposed biosensor could successfully detect the captan and organophosphorus compounds concentration from 0 to 2 ppm.     

Tumor necrosis factor-alpha and its receptor in the corpus luteum of pregnant cows

The objective of this study was to investigate the presence of tumor necrosis factor (TNF)-alpha mRNA and TNF-alpha receptors in the bovine corpus luteum (CL) during the gestation period. TNF-alpha mRNA and TNF-alpha receptors were determined on bovine CL from pregnant cows at three stages: trimester I (fetal crown-rump length; 6-20 cm), trimester II (25-45 cm) and trimester III (50-80 cm). TNF-alpha mRNA was detected by an RT-PCR analysis in the CL of all stages of gestation. A Scatchard analysis revealed the presence of a high-affinity binding site (Kd; 5.1-6.9 nM) in the CL membranes collected at each stage of gestation. Furthermore, the concentrations of TNF-alpha receptors in the CL of trimesters I (24. 0 +/- 1.95 pmol/mg protein) and III (21.6 +/- 2.39 pmol/mg protein) of gestation were significantly higher than the concentration in trimester II (14.9 +/- 2.07 pmol/mg protein) (P < 0.05). These results indicate that TNF-alpha is locally produced and that TNF-alpha receptors are present in bovine CL during the gestation period, and suggest that TNF-alpha plays one or more roles as a paracrine factor in regulating bovine CL function during the entire gestation period.     

Effects of maternal ethanol consumption during pregnancy or lactation on intestinal absorption of folic acid in suckling rats

A fostering/crossfostering analysis of the effects of maternal ethanol exposure on jejunal and ileal folate absorption was performed. Male and female rats were randomized into two groups. In the first group, ethanol-treated rats received ad libitum 5, 10 and 15% ethanol in the drinking fluid during three successive weeks. A consumption of 20% was maintained in this group for 5 additional weeks. Ethanol-treated rats were mated. Group 2 served as the control. To study the effect of chronic alcoholism during lactation or gestation separately, at birth (2nd day postpartum) control newborns were cross-fostered to ethanol dams (EG), and the pups issued from the ethanol treated mothers were cross-fostered to control dams (CG). Thus, three experimental groups of pups were formed: (1) control pups receiving no treatment during gestation and lactation (CG); (2) pups exposed to ethanol only during gestation (GG); and (3) pups exposed to ethanol only during lactation (LG). At 21 days postpartum the jejunal and distal ileum folate absorption was determined in the offspring rats by a perfusion technique. Milk folic acid levels were determined by an immunoluminometric assay. The results showed an increase in jejunal folic acid absorption in offsprings exposed to ethanol only during the lactation period (LG). However, in pups exposed to ethanol only during the gestation period (GG), the jejunal folic acid absorption was significantly increased only at concentrations of 0.25, 0.5 and 2.5 microM. No free folic acid absorption occurred in the distal ileum of control pups (CG) at day 21 at all assayed concentrations but in offsprings exposed to ethanol only during the gestation or lactation periods absorption did take place. Pups exposed to ethanol during the gestation period (GG) showed decreased values in ileum folic acid absorption at the lowest assayed concentration (0.25 microM) compared to values obtained for pups exposed to ethanol only during lactation (LG). Milk folic acid levels were significantly decreased in the ethanol-fed dams on day 21 of lactation. These results indicate that exposure of rats to ethanol during the lactation period affects more severely postnatal development of intestinal functions than ethanol exposure only during gestation. In summary, both the exposure to ethanol itself and the decrease in folic acid intake caused alterations in the function of the intestinal mucosa in the offspring, which in turn altered absorption time and development. However, the present results do not explain how ethanol stimulated intestinal absorption of folic acid in pups exposed to ethanol during the gestation or lactation periods. Further studies are needed.     

Control of tomato grey mould on unheated crops using non- systemic fungicide sprays

High volume sprays of dichlofluanid (0.1 % a.i.) reduced total fruit numbers but gave better control of Botrytis cinerea infections of tomato stems, leaves and fruit than sprays at 0.05 or 0.025% a.i. Surface residues from the sprays at 0–1, 0.05 or 0.025% a.i. were respectively 5.4, 1.6, 0.7 μg/g fresh weight of ripe fruit at harvest. Tank-mixed zineb (0.12% a.i.) and captan (0.2% a.i.) were less effective and increasing the spray intervals from 1 wk to 2 and 3 wk reduced the fungitoxicity of captan and zineb more than that of dichlofluanid. Harvest residues on ripe fruit were 7.1, 2.8 or 2.4 μg/g when dichlofluanid (0.1 % a.i.) sprays were applied at 1, 2 and 3 wk intervals respectively. Good control of B. cinerea was achieved if the whole plant was sprayed initially with dichlofluanid as soon as the second truss was flowering and subsequent sprays were restricted to the upper section of stem including the four or five youngest trusses of buds, flowers and fruit. When used as a post-infection spray there was a period of c. 8 wk before dichlofluanid markedly reduced the incidence of grey mould. Tank-mixed zineb (0.12% a.i.) controlled Botrytis fruit spotting but not leaf and stem infections. Botrytis stem lesions extended more rapidly on zineb-sprayed plants than on unsprayed plants or on plants treated with captan or dichlofluanid.     

Effect of alpha-adrenergic blocking agents on uterine activity in the ewe at the end of gestation

The electromyographic activity (EMG) of the uterus was recorded in vivo in 8 unanaesthetized ewes from the 140th day of gestation up to parturition. The effects on uterine activity of treatments with an alpha 1-receptor blocker (prazosin) and an alpha 2-receptor blocker (yohimbine) were studied. During the last days of gestation, EMG activity consisted of periodic active phases (1-2/h). During the last 16-17 hours, uterine activity increased sharply; this period was referred to a labour. Intravenous perfusion of prazosin (0.03 mg/kg/mn over 1 h) or intravenous injections (1 mg/kg) did not modify uterine activity either before or during labour. Intravenous perfusion of yohimbine (0.03 mg/kg/mn during 1h) inhibited uterine activity before and during labour. In all cases, lambing occurred between the 142nd and 145th day of gestation, which corresponds to the normal lambing period. These results suggest that, in the ewe, uterine alpha 2-receptors are important for normal uterine activity at the end of gestation and in. parturition.     

Studies on the fungitoxicity of captan

The cytological changes in Neurospora crassa conidia following treatment with captan at the ED ₅₀ fungicidal dose have been examined by electron microscopy. With dormant conidia the only observable effect of captan was to produce a characteristic convoluted form of the nuclear membrane. It is suggested that this effect may be due to the reaction of captan with the sulphydryl groups of the nuclear proteins leading to an inhibition of cell division. The cytoplasmic membrane was unaffected by captan, confirming that this fungicide does not destroy cell permeability barriers. After incubation in Fries medium, captan-treated spores showed an almost complete loss of internal fine structure. Similar results were obtained with ‘protoplasts’ of N. crassa which showed disorganization of mitochondrial structure after captan treatment.     

Viruses occurring in white clover (Trifolium repens L.) from permanent pastures in Britain

The uptake of captan from aqueous solution by conidia of Neurospora crassa can be markedly reduced by pretreatment of the cells with thiol reagents such as iodoacetic acid (IOA). The nature of this reaction has been investigated and it is shown that IOA largely reacts with the soluble thiol pool of the spores. Captan, however, reacts with both soluble and insoluble thiols and it is suggested that whilst the former is a detoxication process the latter may provide the key to its toxic action. Using glutathione as a model soluble thiol the molar ratio and pH dependence of the captan detoxication process has been determined and compared with the cellular reactions. Assay of ³⁵S-labelled captan has shown that captan is almost completely decomposed by the spores and that one-third of the accumulated captan is converted to sulphur compounds fixed to insoluble cell entities.     

Successive soil treatment with captan or oxytetracycline affects non-target microorganisms

Changes in microbial biomass and activity were determined in a sandy-loam soil treated with successive dosages of oxytetracycline (a bactericide) or captan (a fungicide) throughout 98 days of incubation under laboratory conditions. The numbers of culturable bacteria and fungi, total bacterial and fungal biomass (as amounts of phospholipid fatty acids, PLFA), the fungal/bacterial ratio, activities of acid and alkaline phosphatases and urease as well as concentrations of N-NH₄ ⁺ and N-NO₃ ⁻ were assessed. Both oxytetracycline and captan significantly decreased numbers of culturable bacteria whereas total bacterial biomass (bactPLFA) was not affected. Oxytetracycline did not effect on the fungal biomass, however their numbers were reduced after the first and second time of soil amendment with the bactericide. Conversely, fungal numbers and biomass (PLFA 18:2ω6,9) significantly decreased in response to soil treatment with the fungicide. Compared to oxytetracycline, captan significantly decreased activities of acid and alkaline phosphatases. For urease activity, the decreased activity was only observed in the soil after the third dosage of captan. Both biocides significantly increased concentrations of N-NH₄ ⁺ and decreased concentrations of N-NO₃ ⁻ after the soil treatments. The results of this study indicate that successive soil treatment with oxytetracycline or captan dosages may negatively affect non-target soil microorganisms and their activities.     

Activity and isoenzyme patterns of glycolytic enzymes during perinatal development of rat lung

The enzymes hexokinase (EC 2.7.1.1), phosphofructokinase (EC 2.7.1.11), enolase (EC 4.2.1.11) and pyruvate kinase (EC 2.7.1.40) were studied in rat lung during development starting at day 16 of gestation (day-6) until 5 days after birth. During gestation, the activities of hexokinase type II, enolase and pyruvate kinase decreased and reached adult values at birth or shortly thereafter. Hexokinase type I remained relatively constant and the decrease of soluble type II hexokinase was compensated for by an increment of particle-bound hexokinase starting at day 20 of gestation until birth. In contrast, phosphofructokinase activity increased until day 20 of gestation followed by a rapid fall in activity until 2 days after birth. Except for hexokinase no isoenzyme shifts were observed in the period of observation. The results are discussed with respect to the proposed relationship between glycogen breakdown and surfactant synthesis during the perinatal period and suggest a regulatory role for phosphofructokinase in this process.     

Alteration of the exonuclease activities of DNA polymerase I by captan

DNA polymerase I is a multifaceted enzyme with one polymerizing and two exonuclease activities. Captan was previously shown to be an inhibitor of this enzyme's polymerizing activity and this report measures the effects of captan on the two exonuclease activities. When the holoenzyme was tested, captan enhanced the degradation of poly(dA-dT), T7 DNA and, to a significantly lesser extent, heat-denatured DNA. However, when the effects of captan were tested as a function of substrate concentration, the stimulatory influence was measured only at high substrate concentrations. At low concentrations of DNA, captan was inhibitory. Inhibition and enhancement each showed an ED50 of the same value (approx. 100 microM). By assaying the two exonuclease activities separately it was shown that the differential effect on the holoenzyme by captan was the result of a combined inhibition of the 3'----5' exonuclease and enhancement of the 5'----3' exonuclease. Klenow fragment with poly(dA-dT) as substrate was used to assay for 3'----5' exonuclease activity. Captan inhibited this exonuclease and the inhibition could be prevented by the addition of greater concentrations of substrate. Holoenzyme and poly(rA)-poly(dT) were used to assay for 5'----3' exonucleolysis, which was enhanced at higher concentrations of substrate in the presence of captan.     

Glasshouse experiments on apple scab

Experiments with fungicide mixtures used protectively and curatively against apple scab showed no advantage from adding phenylmercury chloride (PMC) to certain protective fungicides. Sulphur fungicides greatly impaired the curative activity of PMC, especially in mixtures applied 24 hr. after infection, but captan showed little and dodine acetate no such effect. PMC, however, contributed appreciably to the curative effectiveness of mixtures with captan or dodine acetate. Commercially prepared mixtures of dispersible sulphur with phenylmercury dimethyldithiocarbamate and of zineb with mancozeb gave promising results, and an experimental mixture of dodine/glyodin acetates at low rates was effective protectively and especially curatively. Dodine acetate, dichlofluanid, lime-sulphur, and isobutyl-o-coumarate showed little translocated activity against scab when applied after infection, and PMC, which earlier showed strong translocated activity when applied in summer before infection, was much less effective when applied in spring 24 hr. after infection. Uncontrolled powdery mildew early in the scab-incubation period greatly reduced the establishment of scab infection on the test plants (clonal root-stocks); methods of mildew control in such experiments are suggested.