An efficient protocol for the isolation of fungal DNA for strains obtained from oil contaminated fields

2012 ◽  
Vol 2 (4) ◽  
pp. 162-165
Author(s):  
Bhavya Ravi ◽  
Madhulika Rai ◽  
Sandhya Mehrotra ◽  
Rajesh Mehrotra

A natural ecosystem contaminated with petroleum hydrocarbons is likely to favor the growth of taxonomically diverse microbes having the ability to degrade these organic compounds. They can be exploited for purposes like bioremediation of oil contaminated soils and to obtain enzymes like lipases having important industrial applications. In this paper, a novel “IBG” (Improved ‘Bust and Grab’) protocol has been reported for the isolation of fungal DNA from strains collected from oil contaminated fields. Conventional methods for DNA isolation from fungi require the use of enzymes, liquid nitrogen, glass beads etc. The method reported here circumvents the use of enzymes or glass beads and is cost effective and can be used while handling large number of samples. The DNA yield obtained by the IBG protocol is significant and of good quality. The good quality DNA isolated by IBG protocol can be used for the quick and cost effective isolation of fungal genomic DNA facilitating the genomic study of microbes obtained from oil contaminated fields.

2011 ◽  
Vol 57 (8) ◽  
pp. 623-628 ◽  
Author(s):  
Nagissa Mahmoudi ◽  
Greg F. Slater ◽  
Roberta R. Fulthorpe

Molecular characterization of the microbial populations of soils and sediments contaminated with polycyclic aromatic hydrocarbons (PAHs) is often a first step in assessing intrinsic biodegradation potential. However, soils are problematic for molecular analysis owing to the presence of organic matter, such as humic acids. Furthermore, the presence of contaminants, such as PAHs, can cause further challenges to DNA extraction, quantification, and amplification. The goal of our study was to compare the effectiveness of four commercial soil DNA extraction kits (UltraClean Soil DNA Isolation kit, PowerSoil DNA Isolation kit, PowerMax Soil DNA Isolation kit, and FastDNA SPIN kit) to extract pure, high-quality bacterial and eukaryotic DNA from PAH-contaminated soils. Six different contaminated soils were used to determine if there were any biases among the kits due to soil properties or level of contamination. Extracted DNA was used as a template for bacterial 16S rDNA and eukaryotic 18S rDNA amplifications, and PCR products were subsequently analyzed using denaturing gel gradient electrophoresis (DGGE). We found that the FastDNA SPIN kit provided significantly higher DNA yields for all soils; however, it also resulted in the highest levels of humic acid contamination. Soil texture and organic carbon content of the soil did not affect the DNA yield of any kit. Moreover, a liquid–liquid extraction of the DNA extracts found no residual PAHs, indicating that all kits were effective at removing contaminants in the extraction process. Although the PowerSoil DNA Isolation kit gave relatively low DNA yields, it provided the highest quality DNA based on successful amplification of both bacterial and eukaryotic DNA for all six soils. DGGE fingerprints among the kits were dramatically different for both bacterial and eukaryotic DNA. The PowerSoil DNA Isolation kit revealed multiple bands for each soil and provided the most consistent DGGE profiles among replicates for both bacterial and eukaryotic DNA.


2014 ◽  
Vol 66 (1) ◽  
pp. 285-297 ◽  
Author(s):  
Tanja Dodos ◽  
Jelena Aleksic ◽  
N. Rajcevic ◽  
P.D. Marin

Aromatic species of the genus Satureja are rich in secondary metabolites that interfere with DNA isolation procedures. Four protocols based on the standard CTDNA extraction protocol of Doyle and Doyle (1987) were tested in six savory taxa. The polyphenol adsorbents activated charcoal and/or polyvinylpyrrolidone 10 were employed in three procedures (B, C and D); for the elimination of polysaccharides, 4M NaCl was applied in the latter two. The highest DNA yield was obtained with Protocol D and averaged 1420.7?398.3 ?g DNA/g of dry leaf tissue. Optimal values of the absorbance ratio 260/280 of all DNA solutions revealed the absence or only negligible contamination by proteins. Contamination by polysaccharides inferred from the absorbance ratio 260/230 showed that Protocol C provided the least contaminated material (average of 1.7?0.4). Enzymatic reactions of DNA solutions obtained by Protocol D showed amplification of both loci in all individuals. In conclusion, Protocol D is suitable for the isolation of high quantities of pure DNA from Satureja spp.


Author(s):  
Nikolay S. Shulaev ◽  
◽  
Valeriya V. Pryanichnikova ◽  
Ramil R. Kadyrov ◽  
Inna V. Ovsyannikova ◽  
...  

The most essential scientifific and practical task in the area of ecological safety of pipelines operation is the development and improvement of methods of purifification and restoration of oil-contaminated soils. One of the most effificient and cost effective methods is electrochemical purifification, that does not require the use of expensive chemical reagents and soil excavation. However, the consideration of non-uniform contamination of various soil sections is required. The article examines the features of the organization and technological infrastructure for electrochemical purifification of non-uniformly contaminated soils when using a single electrical energy source, a method for calculating the design parameters of the corresponding installation is proposed. Effificient purifification of non-uniformly contaminated soil when using a specifified voltage is possible through the use of different-sized electrodes. For each soil type, the amount of transmitted electric charge required for soil purifification is determined by the concentration of the contaminant. Allocation of cathodes and anodes as parallel batteries and their connection using individual buses is an effective and energy-effificient solution, since an almost-uniform electric fifield is created in an inter-electrode space, thus allowing the reduction of the interelectrode resistance of the medium.


Sensors ◽  
2021 ◽  
Vol 21 (4) ◽  
pp. 1301
Author(s):  
Federico Cavedo ◽  
Parisa Esmaili ◽  
Michele Norgia

A low-cost optical reflectivity sensor is proposed in this paper, able to detect the presence of objects or surface optical properties variations, at a distance of up to 20 m. A collimated laser beam is pulsed at 10 kHz, and a synchronous digital detector coherently measures the back-diffused light collected through a 1-inch biconvex lens. The sensor is a cost-effective solution for punctual measurement of the surface reflection at different distances. To enhance the interference immunity, an algorithm based on a double-side digital baseline restorer is proposed and implemented to accurately detect the amplitude of the reflected light. As results show, the sensor is robust against ambient light and shows a strong sensitivity on a wide reflection range. The capability of the proposed sensor was evaluated experimentally for object detection and recognition, in addition to dedicated measurement systems, like remote encoders or keyphasors, realized far from the object to be measured.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Shahrbanoo Hamedi ◽  
M. Mehdi Afsahi ◽  
Ali Riahi-Madvar ◽  
Ali Mohebbi

AbstractThe main advantages of the dried enzymes are the lower cost of storage and longer time of preservation for industrial applications. In this study, the spouted bed dryer was utilized for drying the garden radish (Raphanus sativus L.) root extract as a cost-effective source of the peroxidase enzyme. The response surface methodology (RSM) was used to evaluate the individual and interactive effects of main parameters (the inlet air temperature (T) and the ratio of air flow rate to the minimum spouting air flow rate (Q)) on the residual enzyme activity (REA). The maximum REA of 38.7% was obtained at T = 50 °C and Q = 1.4. To investigate the drying effect on the catalytic activity, the optimum reaction conditions (pH and temperature), as well as kinetic parameters, were investigated for the fresh and dried enzyme extracts (FEE and DEE). The obtained results showed that the optimum pH of DEE was decreased by 12.3% compared to FEE, while the optimum temperature of DEE compared to FEE increased by a factor of 85.7%. Moreover, kinetic parameters, thermal-stability, and shelf life of the enzyme were considerably improved after drying by the spouted bed. Overall, the results confirmed that a spouted bed reactor can be used as a promising method for drying heat-sensitive materials such as peroxidase enzyme.


2018 ◽  
Vol 51 (4) ◽  
pp. 291-336 ◽  
Author(s):  
Antimo Graziano ◽  
Shaffiq Jaffer ◽  
Mohini Sain

Blends of polyethylene (PE) and polypropylene (PP) have always been the subject of intense reasearch for encouraging polymer waste recycling while producing new materials for specific applications in a sustainable way. However, being thermodynamically immiscible, these polyolefins form a binary system usually exhibiting lower performances compared with those of the homopolymers. Many studies have been carried out to better understand the PE/PP blend compatibilization for developing a high-performance and cost-effective product. Both nonreactive and reactive compatibilization promote the brittle to ductile transition for a PE/PP blend. However, the final product usually does not meet the requirements for high demanding commercial applications. Therefore, further PE/PP modification with a reinforcing filler, being either synthetic or natural, proved to be a good method for manufacturing high-performance reinforcend polymer blend composites, with superior and tailored properties. This review summarizes the recent progress in compatibilization techniques applied for enhancing the interfacial adhesion between PE and PP. Moreover, future perspectives on better understanding the influence of themodynamics on PE/PP synergy are discussed to introduce more effective compatibilization strategies, which will allow this blend to be used for innovative industrial applications.


2014 ◽  
Vol 71 (9) ◽  
pp. 2530-2538 ◽  
Author(s):  
Emmanuel de Braux ◽  
Fletcher Warren-Myers ◽  
Tim Dempster ◽  
Per Gunnar Fjelldal ◽  
Tom Hansen ◽  
...  

Abstract Otolith marking with enriched stable isotopes via immersion is a recent method of batch marking larval fish for a range of research and industrial applications. However, current immersion times and isotope concentrations required to successfully mark an otolith limit the utility of this technique. Osmotic induction improves incorporation and reduces immersion time for some chemical markers, but its effects on isotope incorporation into otoliths are unknown. Here, we tested the effects of osmotic induction over a range of different isotope concentrations and immersion times on relative mark success and strength for 26Mg:24Mg, 86Sr:88Sr and 137Ba:138Ba on Atlantic salmon (Salmo salar) larvae. 71% and 100% mark success were achieved after 1 h of immersion for 86Sr (75 µg L−1) and 137Ba (30 µg L−1) isotopes, respectively. Compared with conventional immersion, osmotic induction improved overall mark strength for 86Sr and 137Ba isotopes by 26–116%, although this effect was only observed after 12 h of immersion and predominately for 86Sr. The results demonstrate that osmotic induction reduces immersion times and the concentrations of isotope required to achieve successful marks. Osmotically induced isotope labels via larval immersion may prove a rapid and cost-effective way of batch marking fish larvae across a range of potential applications.


Circuit World ◽  
2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Rohollah Abdollahi

Purpose The purpose of this paper is to provide a T autotransformer based 12-pulse rectifier with passive harmonic reduction in more electric aircraft applications. The T autotransformer uses only two main windings which result in volume, space, size, weight and cost savings. Also, the proposed unconventional inter-phase transformer (UIPT) with a lower kVA rating (about 2.6% of the load power) compared to the conventional inter-phase transformer results in a more harmonic reduction. Design/methodology/approach To increase rating and reduce the cost and complexity of a multi-pulse rectifier, it is well known that the pulse number must be increased. In some practical cases, a 12-pulse rectifier (12PR) is suggested as a good solution considering its simple structure and low weight. But the 12PR cannot technically meet the standards of harmonic distortion requirements for some industrial applications, and therefore, they must be used with output filters. In this paper, a 12PR is suggested, which consists of a T autotransformer 12PR and a passive harmonic reduction (PHR) based on the UIPT at direct current (DC) link. Findings To show the advantage of this new combination over other solutions, simulation results are used, and then, a prototype is implemented to evaluate and verify the simulation results. The simulation and experimental test results show that the input current total harmonic distortion (THD) of the suggested 12PR with a PHR based on UIPT is less than 5%, which meets the IEEE 519 requirements. Also, it is shown that in comparison with other solutions, it is cost effective, and at the same time, its power factor is near unity, and its rating is 29.92% of the load rating. Therefore, it is obvious that the proposed rectifier is a practical solution for more electric aircrafts. Originality/value The contributions of this paper are summarized as follows. The suggested design uses a retrofit T autotransformer, which meets all technical constraints, and in comparison, with other options, has less rating, weight, volume and cost. In the suggested rectifier, a PHR based on UIPT at its dc link of 12PR is used, which has good technical capabilities and lower ratings. In the PHR based on UIPT, an IPT is used, which has an additional secondary winding and four diodes. This solution leads to a reduction in input current THD and conduction losses of diodes. In full load conditions, the input line current THD and power factor are 4% and 0.99, respectively. The THD is less than 5%, which satisfies IEEE-519 and DO-160G requirements.


2001 ◽  
Vol 43 (2) ◽  
pp. 291-295 ◽  
Author(s):  
J. Vouillamoz ◽  
M. W. Milke

The effect of compost on phytoremediation of diesel-contaminated soils was investigated using 130 small (200 g) containers in two screening tests. The experiments were conducted in a controlled environment using ryegrass from seed. Containers were destructively sampled at various times and analyzed for plant mass and total petroleum hydrocarbons. The results indicate that the presence of diesel reduces grass growth, and that compost helps reduced the impact of diesel on grass growth. The addition of compost helps increase diesel loss from the soils both with and without grass, though the addition of grass leads to lower diesel levels compared with controls. A second set of experiments indicates that the compost helps in phytoremediation of diesel-contaminated soil independent of the dilution effect that compost addition has. The results indicate that the compost addition allowed diesel loss down to 200 mg TPH/kg even though the compost would be expected to hold the diesel more tightly in the soil/compost mixture. The simplicity of the screening tests led to difficulties in controlling moisture content and germination rates. The conclusion of the research is that the tilling of compost into soils combined with grass seeding appears to be a valuable option for treating petroleum-contaminated soils.


Forests ◽  
2019 ◽  
Vol 10 (6) ◽  
pp. 483 ◽  
Author(s):  
Aleš Ház ◽  
Michal Jablonský ◽  
Igor Šurina ◽  
František Kačík ◽  
Tatiana Bubeníková ◽  
...  

Lignin has great potential for utilization as a green raw material or as an additive in various industrial applications, such as energy, valuable chemicals, or cost-effective materials. In this study, we assessed a commercial form of lignin isolated using LignoBoost technology (LB lignin) as well as three other types of lignin (two samples of non-wood lignins and one hardwood kraft lignin) isolated from the waste liquors produced during the pulping process. Measurements were taken for elemental analysis, methoxyl and ash content, higher heating values, thermogravimetric analysis, and molecular weight determination. We found that the elemental composition of the isolated lignins affected their thermal stability, activation energies, and higher heating values. The lignin samples examined showed varying amounts of functional groups, inorganic component compositions, and molecular weight distributions. Mean activation energies ranged from 93 to 281 kJ/mol. Lignins with bimodal molecular weight distribution were thermally decomposed in two stages, whereas the LB lignin showing a unimodal molecular weight distribution was decomposed in a single thermal stage. Based on its thermal properties, the LB lignin may find direct applications in biocomposites where a higher thermal resistance is required.


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