Comparison of extraction methods for quantifying extracellular polymers in biofilms

1999 ◽  
Vol 39 (7) ◽  
pp. 211-218 ◽  
Author(s):  
Xiaoqi Zhang ◽  
Paul L. Bishop ◽  
Brian K. Kinkle
Keyword(s):  
Extraction Method ◽  
Extracellular Polymeric Substances ◽  
Cell Lysis ◽  
Extraction Methods ◽  
The Other ◽  
Protein Yield ◽  
Total Carbohydrate ◽  
Extracellular Polymers ◽  
Sulfate Reducing ◽  
Edta Extraction

Five commonly used extraction methods - regular centrifugation, EDTA extraction, ultracentrifugation, steaming extraction and regular centrifugation with formaldehyde (RCF) - were selected to study their effectiveness and repeatability in extracting extracellular polymeric substances (EPS) from aerobic/sulfate reducing and nitrifying/denitrifying biofilm samples. Biofilm EPS extraction yields were represented by carbohydrate and protein concentrations; the amount of cell lysis during the extractions was indicated by DNA concentration. The results showed that analyzing wash waters is essential in quantifying biofilm EPS; the contribution of this step varied from 8-50% of the total carbohydrate yield, depending on the extraction method. Among the extraction methods, the RCF extraction gave the greatest carbohydrate yield, the steaming extraction gave the greatest protein yield, and the other three extraction methods gave approximately equivalent amounts of carbohydrate and proteins for both types of biofilm. DNA in the EPS was 27 times smaller than in the pellets, indicating no significant cell lysis occurred during the extractions.

Comparison of Extraction Methods for Quantifying Extracellular Polymers of Marine Algae

2012 ◽  
Vol 260-261 ◽  
pp. 1173-1178 ◽  
Author(s):  
Li Yun Ge ◽  
Yong Gang Huang ◽  
De Xiang Gao ◽  
Huan Huan Deng
Keyword(s):  
Incubation Time ◽  
Extraction Method ◽  
Marine Algae ◽  
Exchange Resin ◽  
Cation Exchange Resin ◽  
Cell Lysis ◽  
Extraction Methods ◽  
Ultrasonic Extraction ◽  
Extracellular Polymers ◽  
Platymonas Subcordiformis

Five extraction methods were used to extract extracellular polymers (EPS) from Platymonas subcordiformis. Results suggested that the extraction efficiencies of EPS decreased as followed: cation exchange resin (CER) > ultrasonic extraction> heating (80°C) > EDTA > centrifugation. CER method was the most effective in the EPS extraction from P. subcordiformis with the highest amount of EPS and the lowest cell lysis, while the ultrasonic extraction method was not suitable to extract EPS for its obvious cell lysis. In addition, heating (80 °C) was used to investigate EPS formation of P. subcordiformis. Results showed EPS amount increased at the increased incubation time when the nutrients were enough whereas the ratio of EPS components remained unchanged in the growing cycle.

The Effects of Solvents and Extraction Methods on the Antioxidant Activity of P. niruri

2014 ◽  
Vol 68 (5) ◽  
Author(s):  
Hafizah Mohd Hadzri ◽  
Mohd Azizi Che Yunus ◽  
Salman Zhari ◽  
Fahim Rithwan
Keyword(s):  
Antioxidant Activity ◽  
Extraction Method ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Soxhlet Extraction ◽  
Solvent Polarity ◽  
Extraction Methods ◽  
The Other ◽  
Natural Antioxidant ◽  
Total Phenolic

The effects of different types of solvents and extraction method were investigated to determine the presence of antioxidant contents and activity from the P. niruri plant. The aim of this study is to determine which extraction method will give higher natural antioxidant contents and antioxidant activity. The content of natural antioxidant and antioxidant activity were analysed by total phenolic content (TPC), total flavonoid content (TFC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity assay. The results showed that extracts from a supercritical fluid extraction (SFE) method without the addition of modifier showed the highest content of total phenolic (187.66 mg GAE/ g) and flavonoid (1100.93 mg QE/g) in P. niruri compared to the other methods of extraction with different type of solvents. The extract of P. niruri from different extraction methods showed antioxidant activity on DPPH radical scavenging assay. The soxhlet extraction method by methanol showed the lowest IC50 compared to the other methods of extraction. The results revealed that P. niruri extracts had different content of antioxidant and antioxidant activity. The solvent polarity and different methods of extraction play significant roles in determining the most suitable method for production of antioxidant contents and antioxidant activity from P. niruri extracts.

Pengambilan senyawa polisakarida ekstraseluler dari mikroorganisme dalam lumpur aktif sebagai adsorben logam berat

2018 ◽  
Vol 5 (2) ◽  
pp. 453
Author(s):  
H Haryono ◽  
Tjandra Setiadi
Keyword(s):  
Heavy Metal ◽  
Activated Sludge ◽  
Extraction Method ◽  
Extracellular Polymeric Substances ◽  
Research Result ◽  
Freundlich Isotherm ◽  
Extraction Methods ◽  
Average Maximum ◽  
Heavy Metal Adsorbent ◽  
Main Component

Extracellular polysaccaride extraction from activated sludge microorganism as heavy-metal adsorbent. Microorganisms in the form of bioagregate are the main component of activated sludge. It generally has an ability to produce EPS (extracellular polymeric substances). The important components in EPS are polysaccharides and proteins. EPS recovery from the activated sludge may be done by many extraction methods. Six of the extraction methods to extract EPS from the activated sludge have been studied in this research, such are: Middle Speed Centrifugation, Regular Centrifugation, NaOH Extraction, EDTA Extraction, Steaming Extraction and Sonication. EPS solution in the following step would be tested its performances as adsorbent of copper heavy metal. The research result shows that the best extraction method for EPS extraction was NaOH Extraction method. The amount of the polysaccharides obtained was 18,09 mg EPS/g TSS. On other hand, Steaming Extraction was the most ineffective extraction method. This method gives the polysaccharides result in the least amount, i.e., about 4,96 mg EPS/g TSS. In this research, the protein content was not detected in the all used EPS solution. In the adsorption test, the adsorption phenomena of copper metal with EPS adsorbent fitted to the Freundlich isotherm adsorption equation. The values of Freundlich's a constant (k and n) each were 2,282 and 0,963. The average maximum adsmption capaciry of EPS to metal Cu was 88,34 mg Cu/gEPS.Keywords:  Adsorption,  Heavy  Metal,  Activated  Sludge,  Extraction Methods, Polysaccharides Extracellular, Copper.AbstrakMikroorganisme dalam bentuk bioagregat adalah merupakan penyusun utama lumpur aktif, memiliki sifat  umum  yaitu  dalam  kemampuannya memproduksi  SPE (Senyawa  Polimer  Ekstraselluler). Komponen  utama dari SPE adalah polisakarida  dan protein. Pengambilan SPE dari bioagregat dapat dilakukan dengan banyak metode ekstraksi.Pada penelitian ini telah dipelajari karakteristik enam  metode  ekstraksi, yaitu: Sentrifugasi Kecepatan Sedang, Sentrifugasi Reguler, Ekstraksi dengan NaOH. Ekstraksi dengan EDTA, Ekstraksi dengan Pemanasan dan Sonikasi. Larutan SPE yang diperoleh, kemudian diuji kinerjanya sebagai adsorben logam berat tembaga. Hasil penelitian menunjukkan bahwa  metode ekstraksi terbaik adalah metode Ekstraksi dengan NaOH, dengan perolehan polisakarida sebesar 18,09 mg/g TSS. Sedangkan Ekstraksi dengan pemanasan merupakan metode ekstraksi yang paling tidak efektif dengan perolehan polisakarida paling sedikit, yaitu sekitar 4,96 mg/g TSS. Pada penelitian ini tidak dijumpai adanya protein di dalam larutan SPE dari semua  metode ekstraksi yang diterapkan. Pada uji adsorpsi, peristiwa adsorpsi logam  Cu dengan larutan SPE lebih mengikuti persamaan adsorpsi isoterm Freundlich dengan harga konstanta k dan n masing-masing sebesar 2,282 dan 0,963. Kapasitas adsorpsi maksimum rata-rata SPE terhadap logam Cu adalah sebesar 88,34 mg Cu/g SPE.Kata kunci:  Adsorpsi, Logam Berat, Lumpur Aktif, Metode Ekstraksi, Polisakarida Ekstraselluler, Tembaga.

An Optimizing Dynamic Spectrum Differential Extraction Method for Noninvasive Blood Component Analysis

2019 ◽  
Vol 74 (1) ◽  
pp. 23-33 ◽  
Author(s):  
Wei Tang ◽  
Qiang Chen ◽  
Wenjuan Yan ◽  
Guoquan He ◽  
Gang Li ◽  
...  
Keyword(s):  
Neural Network ◽  
Correlation Coefficient ◽  
Extraction Method ◽  
Rbf Neural Network ◽  
Extraction Methods ◽  
Dynamic Spectrum ◽  
The Other ◽  
Single Trial ◽  
Differential Extraction ◽  
Better Than

Dynamic spectra (DS) can greatly reduce the influence of individual differences and the measurement environment by extracting the absorbance of pulsating blood at multiple wavelengths, and it is expected to achieve noninvasive detection of blood components. Extracting high-quality DS is the prerequisite for improving detection accuracy. This paper proposed an optimizing differential extraction method in view of the deficiency of existing extraction methods. In the proposed method, the sub-dynamic spectrum (sDS) is composed by sequentially extracting the absolute differences of two sample points corresponding to the height of the half peak on the two sides of the lowest point in each period of the logarithm photoplethysmography signal. The study was based on clinical trial data from 231 volunteers. Single-trial extraction method, original differential extraction method, and optimizing differential extraction method were used to extract DS from the volunteers’ experimental data. Partial least squares regression (PLSR) and radial basis function (RBF) neural network were used for modeling. According to the effect of PLSR modeling, by extracting DS using the proposed method, the correlation coefficient of prediction set ( Rp) has been improved by 17.33% and the root mean square error of prediction set has been reduced by 7.10% compared with the original differential extraction method. Compared with the single-trial extraction method, the correlation coefficient of calibration set ( Rc) has increased from 0.747659 to 0.8244, with an increase of 10.26%, while the correlation coefficient of prediction set ( Rp) decreased slightly by 3.22%, much lower than the increase of correction set. The result of the RBF neural network modeling also shows that the accuracy of the optimizing differential method is better than the other two methods both in calibration set and prediction set. In general, the optimizing differential extraction method improves the data utilization and credibility compared with the existing extraction methods, and the modeling effect is better than the other two methods.

scholarly journals Optimising extraction of extracellular polymeric substances (EPS) from benthic diatoms: comparison of the efficiency of six EPS extraction methods

2009 ◽  
Vol 60 (12) ◽  
pp. 1201 ◽  
Author(s):  
Eri Takahashi ◽  
Jérôme Ledauphin ◽  
Didier Goux ◽  
Francis Orvain
Keyword(s):  
Extracellular Polymeric Substances ◽  
Cell Lysis ◽  
Extraction Methods ◽  
Benthic Diatoms ◽  
Universal Method ◽  
Intertidal Sediments ◽  
Electronic Microscopy ◽  
Glucose Content ◽  
Diatom Culture ◽  
High Yields

There is no universal method that can be applied to extract bound extracellular polymeric substances (EPS) from benthic diatoms of intertidal sediments without causing cell lysis. Six extraction methods were tested on a diatom culture of Navicula jeffreyi to establish the best compromise between high yields of carbohydrate extraction and minimum cell lysis. Extraction with distilled water provoked cell lysis (as already known). The five other extraction methods (dowex resin, artificial seawater of half salinity and extractions after pretreatment with gluteraldehyde by three methods: water, dowex water and dowex buffer) did not provoke cell lysis as shown by transmission electronic microscopy. This result was confirmed by the minimum release of internal compounds (protein, ATP) and by the low proportions of glucose in dowex-extracted EPS compared with the water-extracted EPS, from which the high glucose content must be inferred as contamination by the chrysolaminaran. The extraction with dowex resin resulted in the second-highest concentration of carbohydrate after the water extraction and the EPS were especially rich in deoxy sugars, hence increasing the hydrophobic feature of these substances. For these reasons, we recommend extraction with dowex, which is also the best method for extracting bound EPS from other biofilms such as in activated sludges.

Effective methods for extracting extracellular polymeric substances from Shewanella oneidensis MR-1

2016 ◽  
Vol 74 (12) ◽  
pp. 2987-2996 ◽  
Author(s):  
You-Fen Dai ◽  
Yong Xiao ◽  
En-Hua Zhang ◽  
Li-Dan Liu ◽  
Ling Qiu ◽  
...  
Keyword(s):  
Extracellular Polymeric Substances ◽  
Ethylenediaminetetraacetic Acid ◽  
Heating Temperature ◽  
Three Dimensional ◽  
Shewanella Oneidensis ◽  
Cell Lysis ◽  
Extraction Methods ◽  
Bacterial Cells ◽  
Matrix Spectrum ◽  
Dry Cell

Extracellular polymeric substances (EPS) play crucial roles in bio-aggregate formation and survival of bacterial cells. To develop an effective but harmless method for EPS extraction from Shewanella oneidensis MR-1, five extraction methods, i.e. centrifugation (control), heating (40, 45, 50, and 60 °C), and treatments with H2SO4, ethylenediaminetetraacetic acid (EDTA) and NaOH, were examined, respectively. Results from scanning electron microscope and flow cytometric analyses indicate that MR-1 cells were severely broken by H2SO4, NaOH and heating temperature ≥45 °C. Proteins and polysaccharides in EPS extracted by heating at 40 °C were 7.12 and 1.60 mg g−1 dry cell, respectively. Although EDTA treatment had a relatively lower yield of EPS (proteins and polysaccharides yields of 5.15 and 1.30 mg g−1 dry cell, respectively), cell lysis was barely found after EPS extraction. Three peaks were identified from the three-dimensional excitation–emission matrix spectrum of each EPS sample, suggesting the presence of protein-like substances. Furthermore, the peak intensity was in good accordance with protein concentration measured by the chemical analysis. In short, heating (40 °C) and EDTA treatments were found the most suitable methods for EPS extraction considering the cell lysis and EPS content, composition and functional groups together.

scholarly journals Study on Extracellular Polymeric Substances (EPS) Extraction Method of Lhasa Municipal Sludge

2020 ◽  
Vol 2 (2) ◽  
Author(s):  
Yuhang Fei ◽  
Gaihua Liu ◽  
Lincong Wang ◽  
Jian Lei ◽  
Lei Zhang
Keyword(s):  
Carbon Source ◽  
Extracellular Polymeric Substances ◽  
Extraction Methods ◽  
The Other ◽  
Municipal Sludge ◽  
Sludge Treatment ◽  
Treatment And Disposal ◽  
Main Components ◽  
Microscopic Morphology ◽  
Better Than

Sludge contains large number of EPS and the main components are protein, polysaccharide, hemic acid and so on, which is a great kind of carbon source. Therefore, using a method to extract the EPS and reflux to the biochemical tank can not only provide sufficient carbon source for microorganisms, but also solve the problem of sludge treatment and disposal in sewage plants. This passage compared five different extraction methods to study the extraction effect of EPS of Municipal Sludge. The results showed that the content of EPS components obtained by UN method was higher than that of the other four methods, which indicated that the UN method was better than the other four methods in extracting EPS; Through the comparison and analysis of the SCOD content and microscopic morphology of sludge EPS, it was found that the SCOD content obtained by UN method was the highest, and the corresponding sludge flocculant morphology was destroyed more thoroughly, so the best method was UN method.

scholarly journals Effects of Plant Tissue and DNA Purification Method on Randomly Amplified Polymorphic DNA-based Genetic Fingerprinting Analysis in Carrot

1999 ◽  
Vol 124 (1) ◽  
pp. 32-38 ◽  
Author(s):  
L.S. Boiteux ◽  
M.E.N. Fonseca ◽  
P.W. Simon
Keyword(s):  
Dna Extraction ◽  
Dna Cleavage ◽  
Extraction Method ◽  
Extraction Methods ◽  
The Other ◽  
Dna Purification ◽  
Genetic Fingerprinting ◽  
Purification Method ◽  
Fingerprinting Analysis ◽  
Dna Yield

Seven plant genomic DNA purification protocols were evaluated for genetic fingerprinting analysis using six tissues obtained from inbred carrot (Daucus carota L.) lines. Evaluations included 1) DNA yield, 2) DNA purity, 3) DNA cleavage with HindIII, 4) DNA integrity, and 5) DNA suitability for amplification in a random amplified polymorphic DNA (RAPD) system. Significant differences were observed among tissues and purification methods for the total amount of DNA. An extraction method using CTAB buffer + organic solvents gave the best results in DNA yield, purity, and HindIII cleavage when compared with the other six nonorganic extraction methods. Of the tissues examined, flowers yielded the most DNA (average value = 115 ng of DNA/mg of fresh tissue); followed by seeds (54 ng·mg-1), fresh leaves (48 ng·mg-1), lyophilized leaves (40 ng·mg-1), calli (22 ng·mg-1), and tap roots (4 ng·mg-1). For most of the preparations, the DNA showed no traces of degradation. However, DNA preparations were not consistently accessible to HindIII cleavage in all tissue-extraction method combinations. Uncut DNA was observed chiefly in extractions from flowers and fresh leaves suggesting a tissue-specific adverse effect on restriction endonuclease activity. Differences in RAPD band (amplicon) intensity and number were observed across tissues and DNA extraction methods using identical PCR conditions for RAPD. Callus was the best type of tissue for RAPD-based fingerprinting yielding a consistently higher number of more intense amplicons when compared to the other tissues. In flowers and seeds, only DNA obtained with the CTAB extraction method could be amplified. Polymorphisms deviating from genetic expectations were mainly observed in root and fresh leaf DNA, indicating that some RAPD markers may not present satisfactory levels of reproducibility. Judicious and uniform selection of DNA purification method as well as tissue source for DNA extraction are, therefore, important considerations for reliable RAPD-based DNA fingerprinting analysis in carrot. In addition, our studies allowed the identification of a better combination of procedures for use in routine manipulations of carrot DNA such as RFLP-RAPD-based cultivar fingerprinting, molecular mapping, screening of transgenic plants, construction of genomic libraries, and gene cloning.

Effect of heavy metals (Cu, Pb, and Ni) on the compositions of EPS in biofilms

2001 ◽  
Vol 43 (6) ◽  
pp. 41-48 ◽  
Author(s):  
A. Jang ◽  
S. M. Kim ◽  
S. Y. Kim ◽  
S. G. Lee ◽  
I. S. Kim
Keyword(s):  
Heavy Metals ◽  
Protein C ◽  
Extracellular Polymeric Substances ◽  
Freundlich Isotherm ◽  
Extraction Methods ◽  
Batch Adsorption ◽  
Metal Sorption ◽  
Steam Extraction ◽  
Lead Metal ◽  
Edta Extraction

A series of batch adsorption tests to estimate the biofilm capacity for removal of the heavy metals (Cu, Pb, and Ni) in wastewater were carried out. The metal sorption results were fitted to the Freundlich isotherm model to compare their sorption capacity. The change of the composition of EPSs (extracellular polymeric substances) was simultaneously represented by the ratio of carbohydrate to protein (C/P) when the biofilm was exposed to the heavy metals. EPS composed of slime loosely bound to the cell and capsular materials was extracted by the four general extraction methods including regular centrifugation, regular centrifugation with formaldehyde, EDTA extraction and steam extraction. Although the various extraction methods showed different results, C/P ratio of biofilm exposed to copper and lead metal ions was generally lower than that of control.

scholarly journals A Comparative Study of Contemporary Color Tongue Image Extraction Methods Based on HSI

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Mingfeng Zhu ◽  
Jianqiang Du ◽  
Chenghua Ding
Keyword(s):  
Comparative Study ◽  
Extraction Method ◽  
Active Contour ◽  
Extraction Methods ◽  
Experimental Results ◽  
The Other ◽  
Geodesic Active Contour ◽  
Thick Coating ◽  
Clinical Diagnostic ◽  
Novel Method

Tongue image with coating is of important clinical diagnostic meaning, but traditional tongue image extraction method is not competent for extraction of tongue image with thick coating. In this paper, a novel method is suggested, which applies multiobjective greedy rules and makes fusion of color and space information in order to extract tongue image accurately. A comparative study of several contemporary tongue image extraction methods is also made from the aspects of accuracy and efficiency. As the experimental results show, geodesic active contour is quite slow and not accurate, the other 3 methods achieve fairly good segmentation results except in the case of the tongue with thick coating, our method achieves ideal segmentation results whatever types of tongue images are, and efficiency of our method is acceptable for the application of quantitative check of tongue image.

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