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The main purpose of this study was to examine the influence of pyrethroids, such as deltamethrin, cypermethrin and bifenthrin on the growth and the number of ρ-mutants in the cells of Saccharomyces cerevisiae yeast. SP-4 Mat alpha leu1 arg4 yeast strain was used as a standard strain for experiments. The cells were grown on a standard YPG liquid medium, under aerobic conditions until they reached the logarithmic or stationary phase of growth. Different concentrations of pyrethroid were added to the medium and the cells were incubated for 2 h. The survival rate of the cells was determined by diluting the cells and plating them on YPG Agar plates. The number of ρ-mutants was determined by examining the number of cells that hadn’t grown on YPG medium from among all surviving cells. As far as the investigated forms of pyrethroids are concerned, deltamethrin was the most toxic to yeast cells. It was also observed that low amounts of pyrethroid caused a greater destruction of cells at the logarithmic than at the stationary stage of growth. The influence of pyrethroid on the frequency of mutation of mitochondrial DNA of yeast cells was also studied. It was observed that after incubation with pyrethroid addition the frequency of ρ-mutation increased, especially at the logarithmic stage of growth.
Wyznaczono biodegradacją pyretroidów: cypermetryny, deltametryny, fenwaleratu i permetryny w wodach powierzchniowych średnio zanieczyszczonych. Badania prowadzono w systemie modelowym symulującym warunki wody rzecznej w tzw. modelu rzeki Szybkość procesu degradacji badano w zależności od stężenia związku, temperatury wody i zdolności adaptacyjnych mikroflory wody rzecznej przy powtórnym wprowadzeniu związków do środowiska zanieczyszczonego.
The aim of this work was to determine the activity of proteases and protease inhibitors sampled from the body surface of tau-fluvalinate-sensitive and resistant V. destructor. Proteins were isolated from the tau-fluvalinate-sensitive and resistant mites, while mites untreated with tau-fluvalinate constituted the control. Subsequently, the following methodology was applied: protein concentration assay by the Lowry method — as modified by Schacterle and Pollack; assay of proteolytic activity in relation to various substrates (gelatine, haemoglobin, ovoalbumin, albumin, cytochrome C, casein) by the modified Anson method; identification of proteolytic activity in relation to diagnostic inhibitors of proteolytic enzymes (pepstatin A, PMSF, iodoacetamide, o-phenantrolin), using the Lee and Lin method; identification of acidic, neutral and basic protease activities by means of the modified Anson method; electrophoretic analysis of proteins in a polyacrylamide gel for protease detection with the Laemmli method and for protease inhibitor detection with the Felicioli method. The highest value of protein concentration was found in the tau-fluvalinate-sensitive V. destructor, while the highest activity levels of acidic, neutral and alkaline proteases were observed in the tau-fluvalinate-resistant mites. Aspartic, serine, thiolic and metallic proteases were found in the drug-resistant and drug-sensitive Varroa mites. The control samples were found to contain aspartic and serine proteases. In an acidic and alkaline environment, the results revealed a complete loss of inhibitor activities in the in vitro analyses and electrophoresis. Serine protease inhibitor activities (at pH 7.0) were high, especially in the group of tau-fluvalinate-resistant mites.
Pollen beetle (Meligethes aeneus F.) (PB) is one of the most serious pest in rapeseed cultivations in Poland and in other European countries. The pest is known because of its high metabolic potency towards various toxins. Constant and strong selective pressure of different insecticides used to control this pest, has resulted in resistance of PB to many active substances. In connection with this situation, constant monitoring of pollen beetle susceptibility level to all recommended active substances of insecticides is necessary. The objective of the study was to determine the effectiveness of active substances, most often used in Western Poland for PB control. Resistance of pollen beetle to some pyrethroid active substances and organophosphorous insecticide — phosalone and high susceptibility to chlorpyrifos-ethyl was found. Resistance of pollen beetle requires constant insecticide susceptibility level monitoring and using IRM strategy.
The degradation of chlorophyll a (Chl a), carotenoids and phycobiliproteins, and the fluorescence responses of the filamentous cyanobacterium Anabaena doliolum Bhar. were studied with short exposure (45 min-30 hours) to the pyrethroid insecticide cypermethrin by taking 20 μM and 50 μM of the chemical as treatment concentrations. There was significant reduction in Chl a, carotenoids and phycobiliprotein contents of cells in each of the selected concentrations of the insecticide. Pheophytin a and fluorescing and non-fluorescing chlorophyll catabolites (FCCs and NCCs) were produced as degradation products of Chl a during the exposure period. The amount of the degradation products gradually decreased with prolonged exposure of the cells to the insecticide. The pattern of degradation of carotenoids was similar to that of Chl a but the rate of degradation was less than for Chl a. The amount of lycopene also continuously decreased with increase of the insecticide concentration. There was gradual reduction of cellular phycocyanin and phycoerythrin contents of cells with rapid degradation during the first 6 hours of treatment. The fluorescence emission of phycobilisomes and photosystem II decreased with increased concentrations of insecticide and prolongation of treatment. The cyanobacterium did not show any recovery from insecticide stress during 30 hours of exposure.
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