Ultrasensitive detection of genetically modified maize DNA by capillary gel electrophoresis with laser-induced fluorescence using different fluorescent intercalating dyes     

García-Cañas V  González R  Cifuentes A 《Journal of agricultural and food chemistry》2002,50(16):4497-4502

In this work, four different fluorescent intercalating dyes are compared for the ultrasensitive CGE-LIF detection of DNA from transgenic maize in flours. The fluorescent intercalating dyes compared are YOPRO-1, SYBR-Green-I, Ethidium bromide (EthBr), and EnhanCE. For all the four dyes optimum concentrations are established, and efficient separations of DNA fragments ranging in size from 80 to 1000 bp are obtained. The comparative study demonstrates that SYBR-Green-I and YOPRO-1 provide better limits of detection (LODs) than EnhanCE or EthBr (i.e., LODs are, respectively, 700, 1000, 11300, and 97400 zmol, calculated for a 200-bp DNA fragment). Separations using YOPRO-1 are faster than those using SYBR-Green-I (30 min vs 47 min for the analysis of the 80-1000 bp DNA fragments). Also, separations using YOPRO-1 are more efficient than those using SYBR-Green-I (e.g., 2.4 x 10(6) plates/m vs 1.6 x 10(6) plates/m, respectively, calculated for the 200-bp fragment). Also, buffer depletion and cost per analysis are worse with SYBR-Green-I than with YOPRO-1. Therefore, YOPRO-1 was selected as the preferred intercalating dye. Using this fluorescent compound, analysis time reproducibility for the CGE-LIF separation of the DNA fragments is determined to be better than 1.7% (% RSD, n = 10) within the same day, and better than 1.9% (% RSD, n = 30) for three different days. Moreover, the fluorescence signal obtained using this dye is shown to vary linearly with the DNA concentration in the range studied, i.e., 1-500 ng/microL. It is demonstrated that by using this method 0.01% of transgenic maize can be detected in flour by direct injection of the PCR-amplified sample.  相似文献   

Temporal and Spatial Monitoring of the pH and Heavy Metals in a Soil Polluted by Mine Spill. Post Cleaning Effects   总被引：1，自引：0，他引：1  

Rafaela Ordóñez Fernandez  Juan Vicente Giráldez Cervera  Karl Vanderlinden  Rosa Carbonell Bojollo  Pedro González Fernández 《Water, air, and soil pollution》2007,178(1-4):229-243

The bursting of the mining dam of Aznalcollar (Seville, Spain) triggered an increase in the concentration of heavy metals in the soils of the river Guadiamar valley as a result of the leaching of the pyritic sludge deposited on them. After the cleaning operations which included, as well as mechanical clearing, the addition of different amendments, some areas with residual sludge remained, from which some heavy metals are being mobilized by the cyclical recharge and discharge processes of water in the profiles. This paper analyzes the effect of the soil recovery operations and the climatology on the concentration of metals and their distribution in the soil profile in an area affected by the toxic spill. Fourteen points have been selected in a plot in which acidity persists, there is no vegetation, and residual sludge stains can be seen at a glance. The temporal and spatial evolution of the extractable metals: Fe, Cu, Mn and Zn, the pH and the oxidable fraction has been measured in-depth. The results obtained up to now indicate a leaching of the pollutant towards deeper horizons, finding, at a depth of 757 cm, pH values of 3.5 and very high Fe and Mn concentrations available, especially in the profiles with large sized pores, with a big fraction of sand. On the surface, seasonally, there are low pH values of around 2.5 and extractable Fe contents of over 4000 ppm, which might have an influence on the quality of surface runoff or underground water.  相似文献   

Study of the Levels of Concentration of As, Cd and Ni in a Ceramic Cluster     

S. Pallarés  A. B. Vicente  M. M. Jordán  T. Sanfeliu 《Water, air, and soil pollution》2007,180(1-4):51-64

The main objective of this paper is to check the fulfilment of the European Directive 2004/107/CE, which refers to the limit values of arsenic, cadmium and nickel that will have to be carried out the first of January 2010. Three sampling points have been chosen (Alcora, Vila-real and Castellón), forming a triangle that comprise most of the ceramic cluster of the province of Castellón (Spain). This is a problematic area in relation to the fulfilment of the directive, due to its high industrial development. Apart from this main objective, the following are raised: a) The analysis of the temporal evolution of the levels of As, Cd, Ni and PM10 in the atmosphere during the year 2002; b) The identification of similar behaviour patterns and of the possible common origins in the studied pollutants; c) To show the existence of differences in the behaviour and evolution of As, Cd, Ni and PM10 in the atmospheric medium depending on the location of the sampling point; d) To check whether the levels of As, Cd, Ni and PM10 are influenced by the environmental temperature. The concentration levels of arsenic, cadmium and nickel in Alcora, Vila-real and Castellón have been determined during the year 2002, in order to check whether they are below the limits established in the future directive on these elements. The following conclusions are reached from the results obtained after the chemical analysis (using ICP–MS) of the samples collected in the three locations.  相似文献   

The agronomic benefit of phosphate rock application with elemental sulfur depends on the reactivity and fertilizer placement     

Fábio R. C. F. César  Takashi Muraoka  Rodrigo C. da Silva  Luiz Francisco Souza-Filho 《Journal of plant nutrition》2020,43(18):2773-2784

Abstract

A possible way to improve phosphate rock (PR) agronomic performance is through the addition of elemental sulfur (S⁰). We used ³²P isotope dilution method to assess the P taken up by crops treated with PR. Two experiments, one with common bean and other with upland rice, were carried out, to evaluate the effect of S⁰ on the agronomic performance of two contrasting PR, applied in different methods. Gafsa (GPR) and Patos de Minas (PPR) were used as the high and low reactivity PR, respectively. The experiments were arranged in completely randomized design on factorial structure 2?×?2 × 2?+?2; which means 2 PR (GPR and PPR) versus 2?S⁰ condition (with or without-S⁰) versus 2 application methods (band and broadcast) plus 2 additional treatments (control and triple superphosphate). In band application the S⁰ increased the amount of P uptake by plants from fertilizer of GPR from 2.57 to 9.86?mg pot^?1 for common bean and of 2.26 to 7.05?mg pot^?1 to rice. Regardless the management adopted, less than 2% of P applied as PPR has been taken up by crops. The addition of S⁰ as a strategy to increase the agronomic performance of PR is PR characteristics dependent and fertilizers placement.  相似文献   

Persistent effect of organic matter pulse on a sandy soil of semiarid Patagonia     

Marina González Polo  Esteban Kowaljow  Elisa Castán  Ophelie Sauzet  María Julia Mazzarino 《Biology and Fertility of Soils》2015,51(2):241-249

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Interactive effect between Cu‐adapted arbuscular mycorrhizal fungi and biotreated agrowaste residue to improve the nutritional status of Oenothera picensis growing in Cu‐polluted soils          下载免费PDF全文

Sebastián Meier  Pablo Cornejo  Paula Cartes  Fernando Borie  Jorge Medina  Rosario Azcón 《植物养料与土壤学杂志》2015,178(1):126-135

The interactive effect of sugar beet (SB) agrowaste and arbuscular mycorrhizal (AM) fungi inoculation in response to increasing Cu levels was evaluated in the metallophyte Oenothera picensis. Plants were grown in a Cu‐added soil (0, 100, or 500 mg Cu kg^?1), in presence or absence of SB, and inoculated with: (1) indigenous Cu adapted mycorrhiza (IM) isolated from Cu‐polluted soils; (2) Claroideoglomus claroideum (CC); or (3) maintained uninoculated (control). Sugar beet application produced an increase in shoot biomass of 2 to 7 times, improving plant nutritional status and allowing their survival at the highest Cu concentrations. Moreover, AM fungi utilization had a positive effect promoting the plant establishment; nevertheless, Cu plant concentration as well as the mycorrhizal development in terms of AM colonization, AM spore density, and glomalin production were strictly dependent of the AM fungi strains used. Remarkable differences between AM fungi strains were observed at the highest soil Cu level where only plants colonized by IM were able to survive and grow when no SB residue was added. An interactive effect between AM fungi and SB produced a higher plant growth than plants without the amendment application, improving the plant establishment and allowing their survival at highest copper concentrations, suggesting that this combination could be used as a biotechnological tool for the phytoremediation of Cu‐polluted soils.  相似文献   

Changes in soil nutrient content and bacterial community after 12 years of organic amendment application to a vineyard          下载免费PDF全文

M. E. Calleja‐Cervantes  A. J. Fernández‐González  I. Irigoyen  J. F. Cibriáin‐Sabalza  N. Toro  P. M. Aparicio‐Tejo  M. Fernández‐López 《European Journal of Soil Science》2015,66(4):802-812

An interesting alternative to landfills for disposing of organic residues is their addition to soil as composted organic residues. There is little information available about the long‐term benefits following prolonged periods of application. After 12 years of annual incorporation of organic amendments to the soil of a vineyard, three soil characteristics were analysed: mineral content, bacterial community and soil greenhouse gas (GHG) emissions. The organic amendments were (i) a pelletized organic compost (PEL) made from plant, animal and sewage sludge residues, (ii) a compost made from the organic fraction of municipal solid waste (OF‐MSW) and (iii) a stabilized sheep manure compost (SMC). Mineral fertilizer (NPK) and an unaltered control treatment were also included. Our results showed that long‐term application of treated residues as compost changed soil nutrient content, bacterial community and gas emission rates. For instance, SMC increased nutrients and soil organic matter (OM) throughout the experiment. There was a change in bacterial community structure, with an increase in the phylum Proteobacteria observed for all four treated soils, and an increase in the phylum Bacteroidetes for PEL, OF‐MSW and SMC treatments. Among the organically‐amended soils, the amount of Adhaeribacter increased by a factor of 2.5 times more than the control, which reported a total of 2.0% of the bacterial community compared with 5.6% for PEL, 5.2% for OF‐MSW and 5.0% for SMC. Adhaeribacter may be a genus that specializes in the degradation of residues in the different composts. The SMC treatment had the largest Chao1 estimator and was the most biodiverse of all treatments. These changes in bacterial community structure did not correlate with the observed GHG fluxes from the sampling day. The application of amendments did not affect N₂O fluxes. However, the application of treatments slightly reduced the capacity for CH₄ sequestration by soil with respect to the untreated soils. Compost is an effective method to increase soil fertility. Soil GHG emissions should be further evaluated.  相似文献   

Diagnosis of infectious pancreatic necrosis (IPN) of trout using immunofluorescence     

M Hornich  Z Pospísil  J Tománek 《Veterinární medicína》1986,31(1):55-64

Until recently, demonstration of infectious pancreatic necrosis was conditioned by the isolation of virus. By means of the immunofluorescence method it is possible to make a rapid and accurate diagnosis, as well as to determine the development of infection. In acute cases, the virus in fry was demonstrated mainly in the cytoplasm of epithelial cells of guts; in Pd1-2 trout, the virus was proved in the cytoplasm of epithelial cells of primary kidney tubules and in the cytoplasm of epithelial cells of pancreatic acini. In all cases the virus occurrence demonstrated by immunofluorescence was in correlation with the results achieved by cultivation methods.  相似文献   

Effect of antibiotics, 2-bromoethanesulfonic acid and pyromellitic diimide on methanogenesis in rumen ciliate cultures in vitro.     

Z Váradyová  S Kisidayová  I Zelenák  P Siroka 《Archives of Animal Nutrition》2001,54(1):33-46

The effects of penicillin G, streptomycin, chloramphenicol, 2-bromoethanesulfonic acid and pyromellitic diimide on total gas, methane, volatile fatty acid production and food degradability after 24 h of incubation in vitro were investigated in the cultures of two rumen ciliates. The inocula of both rumen ciliates Entodinium caudatum and Epidinium ecaudatum were used at a volume of 34 ml into the 50 ml glass syringes together with the feed and compounds tested. Despite penicillin G--streptomycin treatment methane production in both cultures was significantly decreased by the inhibitors for Epidinium ecaudatum. Methane production of the bacterial fraction of both protozoan species was significantly lower than in the whole cultures. No epifluorescence of methanogens on (or in) the cells of Entodinium caudatum was observed in contrast to Epidinium with which strong epifluorescence of methanogens on the cell surface was detected. Microscopic observation could indicate that the methane production by Entodinium caudatum was probably caused by their intracellular methanogenic activity, while methane production by Epidinium ecaudatum could be related to both the methanogenic bacterial fraction from their external surface and probably also to intracellular activity. Decreased feed degradability and differences in the fermentation end products accompanied the inhibition of methanogenesis in both in vitro cultures. Entodinium caudatum appeared to be more sensitive than Epidinium ecaudatum to the compounds tested.  相似文献   

The lesional changes and pathogenesis in the kidney in african swine fever     

J. Hervás  J. C. Gómez-Villamandos  A. Méndez  L. Carrasco  M. A. Sierra 《Veterinary research communications》1996,20(3):285-299

African swine fever is a viral haemorrhagic disease of pigs which has been used as a model for the study of viral haemorrhagic diseases in man. The acute course of the disease is characterized by acute proliferative glomerulonephritis, with viral replication in mesangial cells and occasional focal necrosis of the renal tubular system; hyperplasia of the collecting ducts is associated with evident virus replication. Haemorrhages have been attributed to endothelial dysfunction, aggravated by virus replication in endothelial cells in the final stages of the disease. The renal interstitium displays intense oedema and an infiltrate largely composed of macrophages. Virus replication has also been observed in fibroblasts and in the smooth0muscle cells of arterioles and venules.In subacute-chronic forms of the disease, various types of glomerulonephritis are observed, ranging from mesangial proliferative glomerulonephritis to focal and segmental hyalinosis associated with immune-mediated phenomena. No striking changes are reported in the renal tubular system. Interstitial haemorrhages are associated with diapedesis due to immunologically mediated events. The interstitium has also been found to contain a lymphohistiocytic infiltrate with abundant plasma cells. No evidence has been reported of viral replication in any cell population.Abbreviations ASF African swine fever - ASFV African swine fever virus - C complement component - DNA deoxyribonucleic acid - HAD₅₀ 50% haemoadsorbing doses - Ig immunoglobulin - MPS mononuclear phagocytic system - PAS periodic acid-Schiff reagent  相似文献