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A multiple linear regression analyses were performed to screen for the significant factors simultaneously influencing production of delta-endotoxin, proteolytic activities and spore formation by a Bacillus thuringiensis kurstaki strain. Investigated factors included: pH of the medium, available oxygen and inoculum size. It was observed that oxygen availability was the most influencing setting on both deltaendotoxins production and spores counts, followed by initial pH of the medium and inoculum size. On other hand, pH of medium was found to be the most significant parameter for proteolytic activity, followed by inoculum size and dissolved oxygen. Our results suggested that the first order with two-factor interaction model seemed to be more satisfactory than simple first order model for optimization of delta-endotoxin overproduction. The coefficients of determination (R²) indicated a better adequacy of the second order models to justify the obtained data. Based on results, relationships between delta-endotoxins production, proteolytic activities and spores counts were established. Our results can help to balance delta-endotoxins production and its stability.
The study was aimed at estimating species composition and abundance of entomopathogenic fungi in the Chojnowski Landscape park. The effect of site, season (spring, summer, autumn) and temperature on the frequency of isolation of entomopathogenic fungi was also analysed. The effect of the first two factors was estimated based on the analysis of soil samples taken from meadow 1, forest 1 and orchard in spring, summer and autumn 2010. Three species of entomopathogenic fungi (Beauveria bassiana, Metarhizium anisopliae and Isaria fumosorosea) were isolated in the study area. Site and temperature affected the frequency of isolation of particular species. On meadow 1 and in orchard M. anisopliae appeared to be the dominating species while forest 1 was dominated by B. bassiana. From among meadow and forest sites sampled in the autumn, forest 3 (nature reserve) was characterised by the highest density of entomopathogenic fungi. M. anisopliae and B. bassiana were most often isolated from meadow sites while B. bassiana and I. fumosorosea - from forest sites. B. bassiana and I. fumosorosea infected insects with significantly higher frequency at 20°C than at 25°C.
The trials conducted with selected chemical and biological insecticides in 1998-2000 showed the highest effectiveness of Karate Zeon 100 CS (lambda-cyhalotrine) in European corn borer (ECB) larvae control in sweet corn. The efficacy of biological insecticides containing Bacillus thuringiensis ssp. kurstaki: Biobit 3.2 WP and Lepinox WDG was very variable between the years. Reasons for insufficient efficacy of these products are discussed. The most appropriate time for the application of a chemical insecticide against ECB larvae are plant developmental stages since the beginning of pollen shedding to full blooming (63–67 BBCH scale). The efficacy of treatment was the highest at that time.
Effectiveness of 8 bio-rational insecticides, Dipel 2x, BioFly, Agrin, BioGaurd, Spinosad, Neemix, Mectin and Match were tested at recommended dose, half of recommended dose and quarter of recommended dose against 1st, 3rd and 5th instar larvae and the egg masses (24, 48 and 72 h old) of the Egyptian cotton leafworm Spodoptera littoralis (Boisd.) under laboratory conditions. All the bio-rational insecticides provided higher mortality in the first instar larvae comparing to the third and fifth instar larvae, although Match, Mectin and Spinosad showed also excellent efficacy against third larval stage of S. littoralis at all tested concentrations. Also, Match showed 100% mortality of fifth instar larvae at all tested concentrations. The mortality rates of S. littoralis eggs of different ages (24, 48 and 72 h old) when dipped in recommended dose of each insecticide diluted in water were investigated. At recommended dose of all insecticides eggs of different ages were highly affected and the reductions of hatchability were 83.4, 85.0 and 71.7%, respectively in Spinosad compared to the control. In general, eggs 48 and 72 h old were less sensitive than younger eggs 24 h old. The latent effect of bio-rational insecticides on egg hatchability of S. littoralis was observed only in Match and Neemix with the average being 55.0% and 51.6% respectively. Our results suggest that Match, Mectin and Spinosad are potentially effective compounds for control of S. littoralis.
Genetically manipulated food barley, Hordeum vulgare L. var. valfajr, was compared to commercial-grade starch as carbon sources in groundnut and soybean based media for supporting growth, sporulation and delta-endotoxin production by Bacillus thuringiensis Berliner B. thuringiensis kurstaki HD-1 and a promising newly isolated B. thuringiensis strain (referred to as BTA) were used. MgCl2, CaCl2, and MnCl2 were used as trace-elements. Culture media were compared in shaken flasks and then in 5 l Fermentors. Biomass, delta-endotoxin levels and the number of spores as colony forming units (CFU) were evaluated. For each of the two strains, biomass and delta-endotoxin synthesis were not significantly different in soybean-based media after substitution starch by barley flour. There were significant differences between the two strains with respect to biomass and toxin production. Evaluated costs of media preparation showed that food barley is an economical alternative to commercial-grade starch in the production of BTA bioinsecticide.
Highly active antagonistic actinomycète Streptomyces griseoviridis and entomopathogenic fungus Beauveria bassiana were applied to the soil separately and together (in association) in the laboratory experiments. We assessed survival rate, insecticidal and fungistatic activity of these strains. We also tested the influence of synthetic insecticide Regent 25® (fipronil 25g/l) on investigated parameters. Additionally, insecticidal activity of both strains was compared with insecticidal activity of Regent. It was shown that both strains, especially S. griseoviridis, good survived in soil. Population density of S. griseoviridis in 1he association with B. bassiana increased 2-3 times compared to initial density. Regent considerably reduced population density of S. griseoviridis and B. bassiana. Insecticidal efficiency of S. griseoviridis was comparable with the effect of synthetic incecticide Regent and reached 89.2% and 86.8% respectively. Fungistatic activity towards Fusarium oxysporum showed only S. griseoviridis and it was observed that this activity decreased in time course.
The latest developed classes of synthetic insecticides – phthalic acid diamides and antranilic diamides, acting as ryanodine receptor modulators, are discussed. The mode of their activity, the insecticidal properties and characteristics of their representatives is reviewed.
Potential activities of three essential oils (cumin, clove and mustard) and of microbial agents (Paecilomyces fumosoroseus, Nomuraea rileyi, Lecanicillium (Verticillium) lecanii and their combinations against Bruchidius incarnatus (Boh.) were evaluated. In choice test, mustard and clove oils revealed a strong repellent activity after 7 days (89% and 71%, respectively) against B. incarnatus beetles. Cumin oil showed the lowest repellency (47%). Accumulative mortality of beetles increased gradually with the increase of exposure intervals. Mustard oil treatment gave the highest mortality percentage of 76% after 168 h from treatment followed by clove treatment which amounted to 63% and the lowest percentage of 42.8% was recorded in case of cumin oil. Mustard oil was the most effective in enhancing the potency of P. fumosoroseus and N. rileyi and decreased LC50 of the target insect (100 and 102×107, respectively). The persistent effect of formulated mustard oil with either P. fumosoroseus or N. rileyi fungi on foam covering gunny bags displayed several different modes of action, by reducing oviposition and adult emergence (F1) of B. incarnatus. The oviposition was completely inhibited when stored broad bean seeds were treated with mustard oil + P. fumosoroseus during 20, 40 and 60 days of storage. Application of mustard oil combined with P. fumosoroseus on foam covering gunny bags provided promising oviposition deterrency, toxicity and suppressing B. incarnatus infestation, persistence and protecting broad bean seeds from beetles' infestation for 120 days during storage.
A nucleopolyhedrovirus isolated from the beet armyworm Spodoptera exigua (Polish laboratory culture), SeMNPV (P), morphologically similar to the viral bioinsecticide virus Spod-XR , was characterized molecularly and biologically. Phylogenetic analysis based on three conserved baculovirus genes, polh, lef-8 and pif-2, showed the highest homology of SeMNPV (P) to Mamestra brassicae (Mb) MNPV and M. configurata (Maco) MNPV, and much less to SeMNPV (Spod-XR). These findings were confirmed by genomic DNA restriction profile analyses. Bioassays revealed that SeMNPV isolated from the commercial bioinsecticide Spod-XR was themost infectious for S. exigua, while the infectivity of SeMNPV (P) and MbMNPV was significantly lower. These data suggest that SeMNPV (P) is a variant of MbMNPV.
The aim of present study was to determine in vitro susceptibility of the yellow mealworm larvae (Tenebrio molitor, Coleoptera: Tenebrionidae) on some environmental isolates of Bacillus thuringiensis and define the degree of change in the midgut epithelial cells of T. molitor larvae. A study of ultrastructural changes in the midgut of the yellow mealworm larvae treated with B. thuringiensis spores and crystals mixture was carried out. Under laboratory conditions four B. thuringiensis environmental isolates were tested for their insecticidal activity against T. molitor larvae. The tested insects pests showed little susceptibility to bacterial suspensions of spores and crystals (13.3%). Despite the relative pathogenicity the microscopic observation of structure of epithelium cells of T. molitor showed changes in the midgut of the treated larvae.
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