Bacterial reduction of fensulfothion and its hydrolysis product 4-methylsulfinyl phenol.

Oxygen-limited cultures of Klebsiella pneumoniae reduced 4-methylsulfinyl phenol to 4-methylthiophenol. A study of the effect of 4-methylthiophenol on the growth of K. pneumoniae revealed that the specific growth rate was retarded by 40% in the presence of 200 micrograms of the phenol per ml. A soil bacterium, Hafnia sp., was isolated that could reduce the organophosphorus insecticide fensulfothion to fensulfothion sulfide.     

Reduction of fensulfothion to fensulfothion sulfide by Klebsiella pneumoniae.

A cell suspension of Klebsiella pneumoniae converted the organophosphorus pesticide fensulfothion to a product that was shown by chemical oxidation, gas-liquid chromatography, infrared spectrophotometry, and mass spectrometry to be fensulfothion sulfide. Further alteration of this metabolite was not noted.     

Reduction of fensulfothion to fensulfothion sulfide by Klebsiella pneumoniae.

A cell suspension of Klebsiella pneumoniae converted the organophosphorus pesticide fensulfothion to a product that was shown by chemical oxidation, gas-liquid chromatography, infrared spectrophotometry, and mass spectrometry to be fensulfothion sulfide. Further alteration of this metabolite was not noted.     

Metabolism of fensulfothion by a soil bacterium, Pseudomonas alcaligenes C1.

Fensulfothion (O,O-diethyl O-[4-(methylsulfinyl)phenyl]phosphorothioate), an organophosphorus pesticide used to control the golden nematode Heterodera rostochiensis, is used as a source of carbon by microorganisms isolated from soils treated with the pesticide. Two of the microbial isolates, Pseudomonas alcaligenes C1 and Alcaligenes sp. strain NC3, used more than 80% of the pesticide in 120 h in culture when supplemented as a source of carbon. P. alcaligenes C1, which showed maximal growth on fensulfothion, degraded the compound to p-methylsulfinyl phenol and diethyl phosphorothioic acid. The phenolic metabolite could be identified by conventional spectral analysis, whereas the spectral patterns of the phosphorus-containing metabolite suggested that the compound was complexed with some cellular molecules. However, utilization of the phosphoric acid ester and ethanol by P. alcaligenes C1 suggested that the microbe attacks fensulfothion by an initial hydrolysis of the compound and subsequent utilization of the phosphoric acid ester. The pathway of degradation of fensulfothion by P. alcaligenes is of great value in the detoxification of the pesticide residues and also in the environmentally stable phosphoric acid esters.     

Metabolism of fensulfothion by a soil bacterium, Pseudomonas alcaligenes C1

Fensulfothion (O,O-diethyl O-[4-(methylsulfinyl)phenyl]phosphorothioate), an organophosphorus pesticide used to control the golden nematode Heterodera rostochiensis, is used as a source of carbon by microorganisms isolated from soils treated with the pesticide. Two of the microbial isolates, Pseudomonas alcaligenes C1 and Alcaligenes sp. strain NC3, used more than 80% of the pesticide in 120 h in culture when supplemented as a source of carbon. P. alcaligenes C1, which showed maximal growth on fensulfothion, degraded the compound to p-methylsulfinyl phenol and diethyl phosphorothioic acid. The phenolic metabolite could be identified by conventional spectral analysis, whereas the spectral patterns of the phosphorus-containing metabolite suggested that the compound was complexed with some cellular molecules. However, utilization of the phosphoric acid ester and ethanol by P. alcaligenes C1 suggested that the microbe attacks fensulfothion by an initial hydrolysis of the compound and subsequent utilization of the phosphoric acid ester. The pathway of degradation of fensulfothion by P. alcaligenes is of great value in the detoxification of the pesticide residues and also in the environmentally stable phosphoric acid esters.     

Population Dynamics of Belonolaimus longicaudatus and Criconemella ornata and Growth Response of Bermudagrass and Overseeded Grasses on Golf Greens Following Treatment with Nematicides

Portions of a ''Tifgreen'' bermudagrass golf green with poor turf and large numbers of Belonolabnus longicaudatus and Criconemella ornata were treated with selected nematicides in the summers of 1977 and 1978. Improvements in turf quality were observed within 4 wk after treatment with phenamiphos and fensulfothion. Treatment with phenamiphos restulted in lower numbers of B. longicaudatus 4 and 14 wk after treatment in the 1977 experiment and up to 1 yr after treatment in the 1978 experiment. Treatment with fensulfothion reduced the number of B. longicaudatus for only 1 month after treatment and significantly increased the numbers of this nematode in September and March in the 1978 experiment, Negative correlations were obtained between numbers of B. longicaudatus and turf qualily up to 1 yr. Numbers of C. ornata were reduced only in January and June following treatment with phenamiphos and not at any time with fensulfothion. Treatment with fensulfothion resulted in higher numbers of this nematode than in check plots in November and March. The percent area covered by prostrate spurge the following year was reduced following treatment with phenamiphos, but not with fensulfothion.     

Growth kinetics of Pseudomonas putida in cometabolism of phenol and 4-chlorophenol in the presence of a conventional carbon source

Growth kinetics of Pseudomonas putida (ATCC 49451) in cometabolism of phenol and 4-chlorophenol (4-cp) in the presence of sodium glutamate (SG) were studied. In the ternary substrate mixture, phenol and SG are growth substrates while 4-cp is a nongrowth substrate. Cell growth on phenol was found to follow Andrews kinetics and cells displayed substrate inhibition pattern on sodium glutamate in the range of 0-4 g L(-1) as well. A cell growth model for the ternary substrate system was established based on a simplified cell growth mechanism and subsequently modified by experimental results. Model analysis over a wide range of substrate concentrations shows that the inhibition of SG is much larger than phenol at low phenol concentrations (/=600 mg L(-1)). The nongrowth substrate, 4-cp, inhibits cell growth mainly through inactivation of cells (cell decay) and competitive inhibition to cell growth on phenol. In the absence of SG, 4-cp retards cell growth severely and cells cannot grow at 250 mg L(-1) 4-cp. Addition of sodium glutamate, however, greatly attenuates the toxicity of 4-cp and supports cell growth at 4-cp concentration higher than 250 mg L(-1). By using the proposed cell growth model, we were able to optimize the amount of SG needed to enhance cell growth rate and validate model predictions against experimental data.     

Interaction of Fensulfothion and Phorate with Preemergence Herbicides on Soybean Parasitic Nematodes

The herbicides alachlor, linuron, vernolate, and metribuzin were applied to plots treated with the nematicide fensulfothion or the insecticide phorate and planted to soybean in two locations in North Carolina. In 1976 treatment with fensulfothion + alachlor or vernolate, phorate + alachlor or metribuzin resulted in greater nematode population densities than no treatment, or treatment with fensulfothion alone, or phorate alone. In 1977 fensulfothion and phorate alone and in combination with the preemergence herbicides effectively controlled Tylenchorhynchus cIaytoni. Late season population resurgence of Heterodera glycines occurred in fensulfothion + alachlor treated plots. Correlation coefficients for H. glycines vs. yield were -0.48 (P = 0.05) and -0.46 (P = 0.05) for 30 and 68 d after planting, respectively.     

14型肺炎链球菌荚膜多糖纯化工艺中两种去除蛋白方法的比较

目的苯酚抽提法和脱氧胆酸钠沉淀法去除14型肺炎链球菌荚膜多糖中蛋白质的效果比较。方法将3批次14型肺炎链球菌发酵培养液经超滤、乙醇沉淀等方法初步纯化后,平分成两份,分别采用苯酚抽提法和脱氧胆酸钠沉淀法去除蛋白,通过比较多糖收获量、多糖组分检定结果、多糖分子质量、多糖抗原活性、多糖核磁共振图谱,以此评价这两种蛋白去除方法的效果。结果与苯酚抽提法相比,脱氧胆酸钠沉淀法制备的14型肺炎链球菌纯化荚膜多糖除收获量较高,蛋白和核酸杂质含量较低外,氨基己糖含量、多糖分子质量、抗原活性和多糖核磁共振图谱的检定分析结果无显著性差异(P>0.1)。结论作为14型肺炎链球菌荚膜多糖纯化工艺中的除蛋白方法,脱氧胆酸钠沉淀法优于苯酚抽提法。     

Managing Nematode Population Densities on Tomato Transplants Using Crop Rotation and a Nematicide

Millet, milo, soybean, crotalaria, and Norman pigeon pea were used in conjunction with clean fallow and a nematicide (fensulfothion) for managing nematode populations in the production of tomato transplants (Lycopersicon esculentum Mill.). Glean fallow was the most effective treatment in suppressing nematode numbers. After 2 years in tomato, root-knot nematodes increased in numbers to damaging levels, and fallow was no longer effective for complete control even in conjunction with fensulfothion. After 4 years in tomato, none of the crops used as summer cover crops alone or in conjunction with fensulfothion reduced numbers of root-knot nematodes in harvested tomato transplants sufficiently to meet Georgia certification regulations. Milo supported large numbers of Macroposthonia ornata and Pratylenchus spp. and crotalaria supported large numbers of Pratylenchus spp. Millet, milo, soybean, crotalaria, and pigeon pea are poor choices for summer cover crops in sites used to produce tomato transplants, because they support large populations of root-knot and other potentially destructive nematodes.     

Infection of Acanthamoeba castellanii by Chlamydia pneumoniae.

Chlamydia pneumoniae is an intracellular respiratory pathogen, which, similar to Legionella, might have developed mechanisms to escape the intracellular bactericidal activity of both human host cells and amoeba. We therefore investigated the intracellular growth and survival of C. pneumoniae in Acanthamoeba castellanii by using cell culture, immunofluorescence microscopy, and electron microscopy. A castellanii was incubated with purified elementary bodies of C. pneumoniae TW 183 at a concentration of 10(6) inclusion-forming units (IFU)/ml to give a ratio of approximately 1 IFU of C. pneumoniae per amoeba. Quantitative determination of chlamydial growth within A. castellanii revealed viable and infective C. pneumoniae in the range of 10(4) to 10(5) IFU/ml between days 7 and 14 postinfection. Immunofluorescence analysis and transmission electron microscopy with subsequent immunogold staining confirmed evidence of infection of the amoebae by C. Pneumoniae and additionally revealed that C. pneumoniae entered the typical growth cycle. Our results show that amoebae allow the survival of C. pneumoniae, suggesting that amoebae may serve as an additional reservoir for Chlamydia or Chlamydia-related organisms.     

Technetium-99m distribution into Klebsiella pneumoniae.

Bacteria labelled with radionuclide has been the subject of much investigation and has been applied in microbiological research. Technetium-99m (99mTc) may be an alternative radionuclide for the labelling of bacteria employed in various microbiological procedures. This radionuclide is easily available, is not expensive and presents important physical and biological characteristics. 99mTc-labelled bacteria are stable and their cell viability and biological properties are not modified. Study of the distribution of radioactivity in 99mTc-labelled Klebsiella pneumoniae cultures, after homogenization and differential centrifugation of the cells fractions, showed that this radionuclide was present inside the cell, mainly in a ribosomal fraction. Treatment of these fractions with enzymes and detergent revealed a high sensitivity to pronase and Triton X-100. After phenol extraction, a large percentage of radioactivity was detected in the phenol phase. Treatment of the soluble fraction with trichloroacetic acid at different temperatures showed that the concentration of 99mTc in the precipitate was lower at 100 degrees than at 4 degrees C. These results suggest that 99mTc binds mainly to the proteins in Klebsiella pneumoniae.     

Cometabolic degradation of 4-chlorophenol by Alcaligenes eutrophus

 Alcaligenes eutrophus was grown in batch cultures using either phenol as a sole substrate or mixtures of phenol and 4-chlorophenol. Phenol was found to be the sole source for carbon and energy while 4-chlorophenol was utilized only as a cometabolite. Maximum growth rates on phenol reached only 0.26 h^-1, significantly below the growth rates reported earlier with Pseudomonas putida. The cometabolite was found to decrease biomass yield and increase lag time before logarithmic growth occurred. Both phenol and 4-chlorophenol were found to inhibit the growth rate linearly with maximum concentrations of 1080 ppm and 69 ppm respectively, beyond which no growth occurred. The best-fit parameters are incorporated into a simple, dynamic (i.e. time-varying) model capable of predicting all the batch growth conditions presented here. It is shown that P. putida is capable of faster bioremediation when phenol is the sole carbon source or for mixed substrates with low concentrations of the cometabolite, but for high concentrations of 4-chlorophenol, A. eutrophus becomes superior because of the long lag times that occur in the Pseudomonas species. Received: 25 January 1996/Received revision: 13 March 1996/Accepted: 15 April 1996     

Effects of Selected Nematicides on Hatching of Heterodera schachtii

Aldicarb, carbofuran, fensulfothion, and phenamiphos were tested in concentrations of 1-100 μg/ml for their effects on hatching of Heterodera schachtii. Exposure of cysts to 1 μg aldicarb or carbofuran/ml stimulated hatch whereas phenamiphos and, to a lesser degree, fensulfothion inhibited hatch. Addition of aldicarb to sugarbeet root diffusate or 4 mM zinc chloride suppressed activities of these hatching agents. Transfer of cysts previously treated with aldicarb or carbofuran to zinc chloride or water rapidly initiated hatch which finally exceeded the hatch from cysts not treated with the nematicides.     

Biotransformation kinetics of Pseudomonas putida for cometabolism of phenol and 4-chlorophenol in the presence of sodium glutamate

A kinetic model to describe the degradation of phenol and cometabolictransformation of 4-chlorophenol (4-cp) in the presence of sodium glutamate(SG) has been developed and validated experimentally. The integrated modelaccounts for cell growth, toxicity of 4-cp, cross-inhibitions among the threesubstrates, and the different roles of the specific growth substrate (phenol)and the conventional carbon source (SG) in the cometabolism of 4-cp. In thisternary substrate system, the overall phenol degradation and 4-cp transformation rates are greatly enhanced by the addition of SG since SG is able to attenuate the toxicity of 4-cp and therefore increase the cell growth rate. Model analysis indicates that the maximum specific degradation rate of phenol (0.819 mg (mg.h)^-1) is lowered by SG by up to 46% whereas the specific transformation rate of 4-cp is notdirectly affected by the presence of SG. The competitive inhibition coefficient of 4-cp to phenol degradation (K_i,cp) and that of phenol to 4-cp transformation (K_i,ph) were determined to be 6.49 mg l^-1 and 0.193 mg l^-1, respectively, indicatingthat phenol imposes much larger competitive inhibition to 4-cp transformation than the converse. The model developed can simultaneously predict phenol degradation and 4-cp transformation, and is useful for dealing with cometabolism involving multiple substrates.     

Sister-chromatid exchanges and cell-cycle delay in Chinese hamster V79 cells treated with 9 organophosphorus compounds (8 pesticides and 1 defoliant)

Significant increase of sister-chromatid exchanges (SCE) in V79 cells treated with 2 organophosphorus pesticides (OPP), fenthion and oxydemeton-methyl, was observed. The other 7 compounds (6 OPP and 1 defoliant) namely, amaze, azinphos-methyl, bolstar, DEF-defoliant, fensulfothion, monitor and nemacur caused no increase of SCE frequencies at the doses tested. All the compounds except fensulfothion and oxydemeton-methyl induced cell-cycle delay in varying degrees. Cell-cycle delay caused by an OPP was found to be dose-dependent. Based on these data as well as others reported, it would appear that OPP which induce no SCE increase and no or slight cell-cycle delay could be considered as good candidates to substitute the pesticides that have been found to be harmful to the environment.     

异源nifA^C对根癌土壤杆菌nif基因表达的调节作用

用三亲水交配方法分别将载有褐球固氮菌（Ａｚｏｔｏｂａｃｔｅｒｃｈｒｏｏｃｏｃｃｕｍ）呈组成型表达的ｎｉｆＡＣ的质粒ｐＣＫ５和肺炎克氏杆菌（Ｋｌｅｂｓｉｅｌｌａｐｎｅｕｍｏｎｉａｅ）含有ｎｉｆＡ＾Ｃ和ｎｉｆＡ－ｎｔｒＣ基因的质粒ｐＣＫ３,ｐＳＺ３６和ｐＳＺ２３－ＣＡ导入根癌土壤杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍｔｕｎｅｆａｃｉｅｎｓ）Ｃ５８／ｐＧＶ３８５０所得转移接合子的生长速率和野生型相似。在１０ｍ     

Biodegradation of the mixtures of 4-chlorophenol and phenol by Comamonas testosteroni CPW301

A 4-chlorophenol (4-CP)-degrading bacterium, strain CPW301, was isolated from soil and identified as Comamonas testosteroni. This strain dechlorinated and degraded 4-CP via a meta-cleavage pathway. CPW301 could also utilize phenol as a carbon and energy source without the accumulation of any metabolites via the same meta-cleavage pathway. When phenol was added as a additional substrate, CPW301 could degrade 4-CP and phenol simultaneously. The addition of phenol greatly accelerated the degradation of 4-CP due to the increased cell mass. The simultaneous degradation of the 4-CP and phenol is useful not only for enhanced cell growth but also for the bioremediation of both compounds, which are normally present in hazardous waste sites as a mixture.     

Evidence for isofunctional enzymes in the degradation of phenol, m- and p-toluate, and p-cresol via catechol meta-cleavage pathways in Alcaligenes eutrophus.

A study of the degradation of phenol, p-cresol, and m- and p-toluate by Alcaligenes eutrophus 345 has provided evidence that these compounds are metabolized via separate catechol meta-cleavage pathways. Analysis of the enzymes synthesized by wild-type and mutant strains and by strains cured of the plasmid pRA1000, which encodes m- and p-toluate degradation, indicated that two or more isofunctional enzymes mediated several steps in the pathway. The formation of three catechol 2,3-oxygenases and two 2-hydroxymuconic semialdehyde hydrolases was indicated from an examination of the ratio of the specific activities of these enzymes against various substrates. Evidence for two 2-hydroxymuconic semialdehyde dehydrogenases, two 4-oxalocrotonate isomerases and decarboxylases, and three 2-ketopent-4-enoate hydratases was derived from the induction of these enzymes under different growth conditions. Each activity was detected when the wild type was grown in the presence of m-toluate, but not when grown with phenol (except for a hydratase) or p-cresol, whereas in strains cured of pRA1000, growth with phenol or p-cresol, but not with m-toluate, induced these enzymes. Hydroxylation of phenol and p-cresol appears to be mediated by the same enzyme.