Construction of a Recombinant Plasmid from Newcastle Disease Virus Antigen, Haemagglutinin-Neuraminidase and Lactococcal N-Acetylmuramidase and its Expression

The anchoring of proteins to the cell surface of Lactococcus using recombinant DNA techniques is an exciting and emerging research area that holds great promise for a wide variety of biotechnological applications. Presently available anchoring systems are based on recombinant bacteria displaying proteins or peptides on the cell surface. This research is focused on using lactococcal cell wall binding (cwb) region for display of viral epitopes and antigenic determinants. Newcastle disease virus (NDV) is the aetiological agent of Newcastle disease which can result in 100% morbidity and mortality in chicken. Immune response of haemagglutinin-neuraminidase (HN) protein antigens of NDV plays an important role in the prevention of viral infection. A lactococcal domain coding region, acmA’ (300 bp) was cloned into the mammalian expression vector, pcDNA3.1/His. The 1.7 kb HN gene was then inserted at the 5’-end of the acmA’ coding region. Plasmid extraction and restriction enzyme digestion analyses showed that cloning was carried out successfully. Sequencing results indicated that the inserts (acmA’, HN and acmA’HN) were 99%, 97% and 98% homologous to the published sequence of Lactococcus lactis and NDV strain AF2240. The condition for transfection was optimised by testing various amounts of transfected DNA (1 μg, 2 μg and 3 μg) and different charge ratio of liposome reagent (μl): DNA (μg) in a 6-well plate. Transient expression of the fusion protein in Chinese hamster ovary (CHO) cells was analysed by SDS-PAGE and Western blotting. The results indicated that the best combination of DNA used for transfection was 2 μg in 1:3 charge ratio of the liposome:DNA. This study showed that a recombinant acmA’, HN and acmA’-HN have been successfully constructed, transfected and expressed in CHO cells. This work could contribute towards the development of a recombinant vaccine delivery system for NDV

Phenol removal via cellular immobilization: a mini review

Phenolic compounds or phenols are a group of aromatic compounds that comprises a hydroxyl group (OH) that is directly bonded to an aromatic ring. Phenols are injurious to organisms even at even low concentrations with many of them are categorized as dangerous pollutants because of their likely harm to human well-being. This review attempts to discuss the various merits and demerits of immobilization matrices employed for phenol-degrading microorganisms’ immobilization. One of several key points of cellular immobilization is the capacity to protect bioremediation agents towards toxic levels of specific toxicants and safeguarding from predatory microorganisms. However, this shielding course of action should never impede the diffusion of substrates into the pores of the immobilization structure. In the end the choice of a particular immobilization method will strongly hinge on aspects of economy, safety and efficacy

Modelling the Kinetics of Tartrazine Sorption by Bottom Ash

Bottom ash is the solid residue left over from municipal waste combustion or incineration in a Municipal Waste Incineration Furnace. Its use as a sorption agent, particularly for dye sorption, is a new and important application. Linearized adsorption kinetics has drawbacks such as inaccurate representation of the parameters' 95 percent confidence interval output, unbalanced attention to potential outliers, and magnification of errors may result in inaccurate parameter values. In this study, we used nonlinear regression to investigate 16 adsorption kinetics models of tartrazine by bottom ash. The pseudo-second order was the best model based on the Bias and Accuracy factor near unity, but based on other error function analysis, this model performs equally well with the exponential and fractal-like pseudo-second order based other error functions such as Root-Mean-Square Error (RMSE), adjusted coefficient of determination (adjR2), Marquardt’s percent standard deviation (MPSD), Bayesian Information Criterion (BIC), HannanQuinn Information Criterion (HQC), and especially the corrected Akaike Information Criterion (AICc) function as the absolute difference is 5 absolute unit making discriminatory activity difficult. Furthermore, because the pseudo-second order and exponential models have only two parameters, they are less complicated according to Occam's razor. Because the pseudo-second order model is more popular and has more applications than the less well-known exponential model, we chose it as the best model for tartrazine sorption to bottom ash. Kinetic analysis using the PSO model gave a value of equilibrium adsorption capacity, qe of 21.88 mg g-1 (95% confidence interval (C.I.), 20.93 to 22.84) and k2 (g/(mg.sec)) of 0.00002 (95%, C.I., 0.00001 to 0.00002

Screening of hydrocarbon-degrading bacterial isolates using the redox application of 2,6-DCPIP

Petroleum hydrocarbons remain as the major contaminants that could be found across the world. Remediation approach through the utilisation of microbes as the bioremediation means is widely recognised due to their outstanding values. As a result, scientific reports on the isolation and identification of new hydrocarbon-degrading strains were on the rise. Colourimetric-based assays are one of the fastest methods to identify the capability of hydrocarbon-degrading strains in both qualitative and quantitative assessment. In this study, the hydrocarbon-degrading potential of nine bacterial isolates was observed via 2,6-dichlorophenolindophenol (DCPIP) test. Two potent diesel-utilising isolates show a distinctive tendency to utilise aromatic (ADL15) and aliphatic (ADL36) hydrocarbons. Both isolates prove to be a good candidate for bioremediation of wide range of petroleum hydrocarbon components

Kinetic studies on the biosorption of the brominated flame retardant 4-dibromodiphenyl ether (BDE-3) using coconut palm leaf powders

Methods such as membrane isolation, ion replacements, precipitation, transformation and biosorption are proven approaches to contaminant control. Biosorption has all of these technological features including low operating costs, very efficient detoxifying of toxicities at low volumes, minimal amounts of removal components and nutrient requirement, as well as bacterial remediation, which are limited to the presence of heavy metals and other toxicants. The biosorption of BDE-3 on coconut leaves powder on the biosorption of BDE-3 from coconut leaves powder were analyzed using three models—pseudo-1st, pseudo-2nd and Elovich, and fitted using non-linear regression. Statistical analysis based on root-mean-square error (RMSE), adjusted coefficient of determination (adjR2), bias factor (BF), accuracy factor (AF), corrected AICc (Akaike Information Criterion), Bayesian Information Criterion (BIC) and Hannan–Quinn information criterion (HQC) showed that the Pseudo-2nd order model is the best model. Kinetic analysis using the Pseudo-2nd order model gave a value of equilibrium sorption capacity qe for 0.01 g per L adsorbent of 488.16 mg g-1 (95% confidence interval from 463.68 to 512.64) and a value of the Pseudo-2nd-order rate constant, k2 of 0.00019 (95% confidence interval from 0.00010 to 0.00027) while the equilibrium sorption capacity qe for 0.002 g per L adsorbent of 2403.61 mg g-1 (95% confidence interval from 2313.99 to 2493.22) and a value of the Pseudo-2nd-order rate constant, k2 of 0.000043 (95% confidence interval from 0.000027 to 0.000059). These calculated values will be very useful in designing effective sorption experiment and understanding the limitations of the system developed

Environmental Fate and Degradation of Glyphosate in Soil

oai:ojs1.pjsrr.upm.edu.my:article/124Commercialisation of glyphosate [N-(phosphonomethyl)glycine] in the early 1970s has left a big leap in the agriculture sector. This is due to its effectiveness in controlling a wide range of weeds. Glyphosate translocates well in plants. In addition, with added surfactant in its formulae, it can also be used in wet conditions. Its ability to kill weeds by targeting the 5-enolpyruvyl-shikimate-3-phosphate synthase (EPSPS) makes no competing herbicide analogs in its class. Considering its cost effectiveness, only small amount is needed to cover a large sector in agricultural land. The most important aspect in the success of glyphosate is the introduction of transgenic, glyphosate-resistant crops in 1996. However, glyphosate is not an environmental friendly herbicide. This systematic herbicide has raised environmental concern due to its excessive use in agriculture. Studies have shown traces of glyphosate found in drinking water. Meanwhile, it's rapid binding on soil particles possesses adverse effect to soil organisms. Glyphosate degradation in soil usually carried out by microbial activity. Microbes’ capable utilising glyphosate mainly as phosphate source. However, the activity of C-P lyase in breaking down glyphosate have not clearly understood. This review presents a collective summary on the understanding on how glyphosate works and its environmental fate

Bioremediation of petroleum hydrocarbon in Antarctica by microbial species: an overview

The increase of anthropogenic activities and growth of technology in Antarctica is fuelled by the high demand for petroleum hydrocarbons needed for daily activities. Oil and fuel spills that occur during explorations have caused hydrocarbon pollution in this region, prompting concern for the environment by polar communities and the larger world community. Crude oil and petroleum hydrocarbon products contain a wide variety of lethal components with high toxicity and low biodegradability. Hydrocarbon persistence in the Antarctic environment only worsens the issues stemming from environmental pollution as they can be long-term. Numerous efforts to lower the contamination level caused by these pollutants have been conducted mainly in bioremediation, an economical and degrading-wise method. Bioremediation mainly functions on conversion of complex toxic compounds to simpler organic compounds due to the consumption of hydrocarbons by microorganisms as their energy source. This review presents a summary of the collective understanding on bioremediation of petroleum hydrocarbons by microorganisms indigenous to the Antarctic region from past decades to current knowledge

Isothermal modelling of the adsorption of lead(II) onto an antarctic sea-ice bacterial exopolysaccharide

The biosorption of the biosorption of lead(II) onto an Antarctic sea-ice bacterial exopolysaccharide is remodeled using nonlinear regression and the optimal mode was determined by a series of error function assessments. The Sips model performed best in statistical tests including root-mean-square error (RMSE), adjusted coefficient of determination (adjR2), bias factor (BF), accuracy factor (AF), and corrected Akaike Information Criterion (AICc) which is not the same to the originally published work using a linearized form where the Langmuir and Freundlich models best represent the biosorption and the maximum biosorption capacity. The calculated Sips parameters kS (l/g) value of 0.10 (95% confidence interval from 0.08 to 0.13), a maximum monolayer adsorption capacity qmS (mg/g) value of 252.88 (95% C.I. from 222.13 to 283.64) and nS (Sips model exponent) value of 1.16 (95% C.I. from 1.34 to 1.98). This study indicates that a different isotherm model can be obtained using nonlinear regression compared to the popular linearized form that may give relatively inaccurate outcome

Assay for heavy metals using an inhibitive assay based on the acetylcholinesterase from Clarias batrachus

Acetylcholinesterase (AChE) is usually used as an inhibitive assay for insecticides. A lesser-known property of AChE is its inhibition by heavy metals. In this work, we evaluate an AChE from brains of Clarias batrachus (catfish) exposed to wastes from aquaculture industry as an inhibitive assay for heavy metals. We discovered that the AChE was inhibited completely by Hg2+, Ag2+, Pb2+, Cu2+, Cd2+, Cr6+ and Zn2+ during initial screening. When tested at various concentrations, the heavy metals exhibited exponential decay type inhibition curves. The calculated IC50 (mg/L) for the heavy metals Ag2+, Cu2+, Hg2+, Cr6+ and Cd2+ were 0.088, 0.078, 0.071, 0.87 and 0.913, respectively. The IC50 for these heavy metals are comparable, and some are lower than the IC50 values from the cholinesterases from previously studied fish. The assay can be carried out in less than 30 minutes at ambient temperature

Kinetic analysis of the adsorption of lead (II) onto an antarctic sea-ice bacterial exopolysaccharide

Hypertension and kidney impairment are two of the many adult health problems that have been related to lead exposure. Women who are expecting a child are especially susceptible to the dangers of lead since it can have devastating consequences on the developing embryo. Existing techniques for the remediation of lead pollutant include membrane separation, ion exchange, precipitation and biosorption. Of all of this technology, biosorption has several positive aspects which include low operating expenses, very efficient detoxification of toxicants at low concentrations and low amount of disposal materials. The biosorption of the biosorption of lead(II) onto an Antarctic sea-ice bacterial exopolysaccharide is remodeled using nonlinear regression and the optimal mode was determined by a series of error function assessments. Statistical analysis showed that the best kinetic model for adsorption in salt-free water was pseudo-1st order while the best kinetic model for adsorption in seawater was pseudo-2nd order model. All error function analyses also supported these two best models. The kinetic constants values for salt-free water and seawater shows large difference in terms of adsorption in salt-free water and seawater. A higher equilibrium biosorption capacity for lead (II) or qe values were exhibited for both k1 and k2 rate constants in sea water indicating a more efficient adsorption in seawater. Adsorption in seawater increased the qe values from 51.11 (mg/g) (95% confidence interval from 49.75 to 52.44) to 92.98 (mg/g) (95% C.I. from 91.01 to 94.95) In addition, the h value, (mg/g.min) indicates a stronger driving force to accelerate the diffusion of adsorbate from seawater onto the adsorbent. The results suggest fundamental difference of sorption mechanism and functional groups are involved in salt-free and seawater