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Fulltext Glyphosate Herbicide Induces Changes in the Growth Pattern and Somatic Indices of Crossbred Red Tilapia (O. niloticus × O. mossambicus)

Muhammad UA, Yasid NA, Daud HM, Shukor MY

Animals (Basel), 2021 Apr 22;11(5).
PMID: 33922293 DOI: 10.3390/ani11051209

The development of glyphosate-resistant genetically modified organisms (GMO) has increased the use of herbicide glyphosate by several magnitudes in recent years. It is now the most commonly used pesticide globally that affects aquatic habitats, especially fish. This study aims to add new knowledge on the effect of technical grade glyphosate on several toxicity parameters and to identify the most effective parameter in predicting technical grade glyphosate chronic toxicity (seven weeks) to fish, especially Malaysia's heavily farmed red tilapia. The results show that a relatively high concentration of technical grade glyphosate is needed to induce significant changes in all tested parameters. However, the results also indicate that the bodyweight index is the most sensitive toxicity parameter in that a reduction in body weight was observed at 25 mg/L of glyphosate. Negative correlations between the glyphosate concentration and toxicity parameters such as specific growth rate (SGR), hepato-somatic index (HIS), and gonado-somatic index (GSI) were observed. The fish condition factor and feed conversion ratio were found not to be affected at the highest glyphosate concentration tested (150 mg/L). To conclude, crossbred red tilapia (O. niloticus × O. mossambicus) is one potential species for evaluating the toxic effects of technical grade glyphosate on fish.

Matched MeSH terms: Organisms, Genetically Modified
Fulltext Review paper : application of DNA and immunoassay analytical methods for GMO testing in agricultural crops and plant-derived products

Jasbeer, K., Ghazali, F.M., Cheah, Y.K., Son, R.

International Food Research Journal, 2008;15(1):1-25.
MyJurnal

The introduction of new agricultural commodities and products derived from modernbiotechnology may have an impact on human and animal health, the environment and economiesof countries. As more Genetically Modified Organisms (GMO) enter markets worldwide, themonitoring of GMOs is being preferred for obvious reasons such as determination of seed purity,verification of non-GMO status of agricultural crops and fulfilling GMO labeling provisions, tomention a few. Numerous GMO analytical methods which include screening, identification andquantification have been developed to reliably determine the presence and/or amount of GMOin agricultural commodities, in raw agricultural materials and in processed and refined ingredients.The detection of GMOs relies on the detection of transgenic DNA or protein material. For routineanalysis, a good sample preparation technique should reproducibly generate DNA/protein ofsufficient quality, purity and yield while minimizing the effects of inhibition andcontamination.
The key sample preparation steps include homogenization, pretreatment, extraction andpurification. Due to the fact that analytical laboratories receive samples that are often processedand refined, the quality and quantity of transgenic target analyte (e.g. protein and DNA) frequentlychallenge the sensitivity of any detection method. With the development of GMO analysistechniques, the Polymerase Chain Reaction (PCR) technique has been the mainstay for GMOdetection, and the real-time PCR is the most effective and important method for GMOquantification. The choice of target sequence; for example a promoter, a terminator, a gene, or ajunction between two of these elements, is the single most important factor controlling the specificity of the PCR method. Recent developments include event-specific methods, particularlyuseful for identification and quantification of GM content. Although PCR technology has obvious
limitations, the potentially high degree of sensitivity and specificity explains why PCR in its various
formats, is currently the leading analytical technology employed in GMO analysis. Comparatively, immunoassays are becoming attractive tools for rapid field monitoring for the integrity of agricultural commodities in identity preservation systems, whereby non-specialised personnel can employ them in cost-effective manner. This review discusses various popular extraction methodologies and summarises the current status of the most widely used and easily applicable GMO analysis technologies in laboratories, namely the PCR and immunoassay technologies.

Matched MeSH terms: Organisms, Genetically Modified
Surface plasmon resonance biosensor for real-time detection of genetically modified organisms

Yoke-Kqueen, C., Son, R.

International Food Research Journal, 2010;17(2):477-483.
MyJurnal

Application of surface plasmon resonance (SPR) biosensor in detection of genetically modified organism (GMO) is demonstrated. A total of four biotinylated probes namely Tnosb, P35Sb, LECb and TSQb were successfully immobilized onto the SA chip. Results analysis indicated that the SPR system with the sensor chip immobilized with the Tnosb, P35Sb, LECb and TSQb biotinylated probes potentially detect complementary standard fragments as low as 1 nM. Biospecific interaction analysis (BIA), employing surface plasmon resonance (SPR) and biosensor technologies provide easy, rapid and automatable approach in detection of GMOs. Short assay times, label free DNA hybridization reaction and no toxic compounds are required, i.e. ethidium bromide, and the reusability of the sensor surface are some of the factors that contribute to the general advantages of the surface plasmon resonance (SPR) biosensor system in detection of GMOs.

Matched MeSH terms: Organisms, Genetically Modified
Natural resources management and food security in the context of sustainable development

Hill J, Saleem Mustafa

Sains Malaysiana, 2011;40:1331-1340.

This paper elaborates on the inseparable link between sustainability of natural resources and food security. A strategic framework that envisages conservation, improvement and sustainable uses of natural resources is proposed which meets the essential requirements for food security. Sustainability has traditionally been accepted as encompassing three dimensions, namely environment, economics and society but it is necessary to widen this approach for a more complete understanding of this term. Environmental degradation curtails ecosystem services, leading to impoverishment of vulnerable communities and insecurity. Food, whether derived from land or sea, is a product of complex environmental linkages, and biodiversity has a pivotal role to play in producing it. Technology, production methods and management requirements are different for food derived from land and sea, but essentially all foodstuffs utilize environmental resources whose sustainability is crucial for food security. This analysis necessitates consideration of the basic concepts of sustainable development and food security, the strength of the link between these and differences in the patterns of sustainable management of agriculture, fisheries and aquaculture. The growing role of genetically engineered organisms has been included because of the immense possibilities these offer for maximizing food production despite the environmental and ethical concerns raised.

Matched MeSH terms: Organisms, Genetically Modified
Factors influencing stakeholders attitudes toward genetically modified aedes mosquito

Amin L, Hashim H

Sci Eng Ethics, 2015 Jun;21(3):655-81.
PMID: 24906652 DOI: 10.1007/s11948-014-9557-5

Dengue fever is a debilitating and infectious disease that could be life-threatening. It is caused by the dengue virus which affects millions of people in the tropical area. Currently, there is no cure for the disease as there is no vaccine available. Thus, prevention of the vector population using conventional methods is by far the main strategy but has been found ineffective. A genetically modified (GM) mosquito is among the favoured alternatives to curb dengue fever in Malaysia. Past studies have shown that development and diffusion of gene technology products depends heavily upon public acceptance. The purpose of this study is to identify the relevant factors influencing stakeholders' attitudes toward the GM Aedes mosquito and to analyse the relationships between all the factors using the structural equation model. A survey was carried out on 509 respondents from various stakeholder groups in the Klang Valley region of Malaysia. Results of the survey have confirmed that public perception towards complex issues such as gene technology should be seen as a multi-faceted process. The perceived benefit-perceived risk balance is very important in determining the most predominant predictor of attitudes toward a GM mosquito. In this study the stakeholders perceived the benefit of the GM mosquito as outweighing its risk, translating perceived benefit as the most important direct predictor of attitudes toward the GM mosquito. Trust in key players has a direct influence on attitudes toward the GM mosquito while moral concern exhibited an indirect influence through perceived benefits. Other factors such as attitudes toward technology and nature were also indirect predictors of attitudes toward the GM mosquito while religiosity and engagement did not exhibited any significant roles. The research findings serve as a useful database to understand public acceptance and the social construct of public attitudes towards the GM mosquito to combat dengue.

Matched MeSH terms: Organisms, Genetically Modified*
Letting the bugs out of the bag

Nature, 2011 Feb 10;470(7333):139.
PMID: 21307891 DOI: 10.1038/470139a
Matched MeSH terms: Organisms, Genetically Modified*
Fulltext Dispersal of Engineered Male Aedes aegypti Mosquitoes

Winskill P, Carvalho DO, Capurro ML, Alphey L, Donnelly CA, McKemey AR

PLoS Negl Trop Dis, 2015 Nov;9(11):e0004156.
PMID: 26554922 DOI: 10.1371/journal.pntd.0004156

BACKGROUND: Aedes aegypti, the principal vector of dengue fever, have been genetically engineered for use in a sterile insect control programme. To improve our understanding of the dispersal ecology of mosquitoes and to inform appropriate release strategies of 'genetically sterile' male Aedes aegypti detailed knowledge of the dispersal ability of the released insects is needed.
METHODOLOGY/PRINCIPAL FINDINGS: The dispersal ability of released 'genetically sterile' male Aedes aegypti at a field site in Brazil has been estimated. Dispersal kernels embedded within a generalized linear model framework were used to analyse data collected from three large scale mark release recapture studies. The methodology has been applied to previously published dispersal data to compare the dispersal ability of 'genetically sterile' male Aedes aegypti in contrasting environments. We parameterised dispersal kernels and estimated the mean distance travelled for insects in Brazil: 52.8 m (95% CI: 49.9 m, 56.8 m) and Malaysia: 58.0 m (95% CI: 51.1 m, 71.0 m).
CONCLUSIONS/SIGNIFICANCE: Our results provide specific, detailed estimates of the dispersal characteristics of released 'genetically sterile' male Aedes aegypti in the field. The comparative analysis indicates that despite differing environments and recapture rates, key features of the insects' dispersal kernels are conserved across the two studies. The results can be used to inform both risk assessments and release programmes using 'genetically sterile' male Aedes aegypti.

Matched MeSH terms: Organisms, Genetically Modified*
Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxy-4-methylvalerate) by recombinant Escherichia coli expressing leucine metabolism-related enzymes derived from Clostridium difficile

Saika A, Watanabe Y, Sudesh K, Tsuge T

J Biosci Bioeng, 2014 Jun;117(6):670-5.
PMID: 24484910 DOI: 10.1016/j.jbiosc.2013.12.006

An obligate anaerobic bacterium Clostridium difficile has a unique metabolic pathway to convert leucine to 4-methylvalerate, in which 4-methyl-2-pentenoyl-CoA (4M2PE-CoA) is an intermediate of this pathway. 4M2PE-CoA is also able to be converted to 3-hydroxy-4-methylvalerate (3H4MV), a branched side chain monomer unit, for synthesis of polyhydroxyalkanoate (PHA) copolymer. In this study, to synthesize 3H4MV-containing PHA copolymer from leucine, the leucine metabolism-related enzymes (LdhA and HadAIBC) derived from C. difficile and PHA biosynthesis enzymes (PhaPCJAc and PhaABRe) derived from Aeromonas caviae and Ralstonia eutropha were co-expressed in the codon usage-improved Escherichia coli. Under microaerobic culture conditions, this E. coli was able to synthesize P(3HB-co-12.2 mol% 3H4MV) from glucose with the supplementation of 1 g/L leucine. This strain also produced P(3HB-co-12.6 mol% 3H4MV) using the culture supernatant of leucine overproducer E. coli strain NS1391 as the medium for PHA production, achieving 3H4MV copolymer synthesis only from glucose. Furthermore, we tested the feasibility of the 3H4MV copolymer synthesis in E. coli strain NS1391 from glucose. The recombinant E. coli NS1391 was able to synthesize P(3HB-co-3.0 mol% 3H4MV) from glucose without any leucine supplementation. This study demonstrates the potential of the new metabolic pathway for 3H4MV synthesis using leucine metabolism-related enzymes from C. difficile.

Matched MeSH terms: Organisms, Genetically Modified/genetics; Organisms, Genetically Modified/metabolism
Recovery and purification of intracellular polyhydroxyalkanoates from recombinant Cupriavidus necator using water and ethanol

Mohammadi M, Hassan MA, Phang LY, Ariffin H, Shirai Y, Ando Y

Biotechnol Lett, 2012 Feb;34(2):253-9.
PMID: 22038551 DOI: 10.1007/s10529-011-0783-5

A new halogen-free and environmental-friendly method using water and ethanol is developed as an alternative for the recovery of polyhydroxyalkanoates (PHA) from recombinant Cupriavidus necator in comparison to the established chloroform extraction method. After optimisation, our results showed that the halogen-free method produced a PHA with 81% purity and 96% recovery yield, in comparison to the chloroform extraction system which resulted in a highly pure PHA with 95% yield. Although the purity of the PHA using the new method is lower, the molecular weight of the extracted PHA is not compromised. This new method can be further developed as an alternative and more environmental-friendly method for industrial application.

Matched MeSH terms: Organisms, Genetically Modified
Cell surface display system for Lactococcus lactis: a novel development for oral vaccine

Raha AR, Varma NR, Yusoff K, Ross E, Foo HL

Appl Microbiol Biotechnol, 2005 Jul;68(1):75-81.
PMID: 15635459

The food-grade Lactococcus lactis is a potential vector to be used as a live vehicle for the delivery of heterologous proteins for vaccine and pharmaceutical purposes. We constructed a plasmid vector pSVac that harbors a 255-bp single-repeat sequence of the cell wall-binding protein region of the AcmA protein. The recombinant plasmid was transformed into Escherichia coli and expression of the gene fragment was driven by the T7 promoter of the plasmid. SDS-PAGE showed the presence of the putative AcmA' fragment and this was confirmed by Western blot analysis. The protein was isolated and purified using a His-tag affinity column. When mixed with a culture of L. lactis MG1363, ELISA and immunofluorescence assays showed that the cell wall-binding fragment was anchored onto the outer surface of the bacteria. This indicated that the AcmA' repeat unit retained the active site for binding onto the cell wall surface of the L. lactis cells. Stability assays showed that the fusion proteins (AcmA/A1, AcmA/A3) were stably docked onto the surface for at least 5 days. The AcmA' fragment was also shown to be able to strongly bind onto the cell surface of naturally occurring lactococcal strains and Lactobacillus and, with less strength, the cell surface of Bacillus sphericus. The new system designed for cell surface display of recombinant proteins on L. lactis was evaluated for the expression and display of A1 and A3 regions of the VP1 protein of enterovirus 71 (EV71). The A1 and A3 regions of the VP1 protein of EV71 were cloned upstream to the cell wall-binding domains of AcmA protein and successfully expressed as AcmA/A1 and AcmA/A3. Whole-cell ELISA showed the successful display of VP1 protein epitopes of EV71 on the surface of L. lactis. The success of the anchoring system developed in this study for docking the A1 and A3 epitopes of VP1 onto the surface of L. lactis cells opens up the possibilities of peptide and protein display for not only Lactococcus but also for other gram-positive bacteria. This novel way of displaying epitopes on the cell surface of L. lactis and other related organisms should be very useful in the delivery of vaccines and other useful proteins.

Matched MeSH terms: Organisms, Genetically Modified
Effect of signal peptides on the secretion of β-cyclodextrin glucanotransferase in Lactococcus lactis NZ9000

Subramaniam M, Baradaran A, Rosli MI, Rosfarizan M, Khatijah Y, Raha AR

J. Mol. Microbiol. Biotechnol., 2012;22(6):361-72.
PMID: 23295307 DOI: 10.1159/000343921

Cyclodextrin glucanotransferase (CGTase) is an extracellular enzyme which catalyzes the formation of cyclodextrin from starch. The production of CGTase using lactic acid bacterium is an attractive alternative and safer strategy to produce CGTase. In this study, we report the construction of genetically modified Lactococcus lactis strains harboring plasmids that secrete the Bacillus sp. G1 β-CGTase, with the aid of the signal peptides (SPs) SPK1, USP45 and native SP (NSP). Three constructed vectors, pNZ:NSP:CGT, pNZ:USP:CGT and pNZ:SPK1:CGT, were developed in this study. Each vector harbored a different SP fused to the CGTase. The formation of halo zones on starch plates indicated the production and secretion of β-CGTase by the recombinants. The expression of this enzyme is shown by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and zymogram analysis. A band size of ∼75 kDa corresponding to β-CGTase is identified in the intracellular and the extracellular environments of the host after medium modification. The replacement of glucose by starch in the medium was shown to induce β-CGTase production in L. lactis. Although β-CGTase production is comparatively low in NZ:SPK1:CGT, the SP SPK1 was shown to have higher secretion efficiency compared to the other SPs used in this study.

Matched MeSH terms: Organisms, Genetically Modified
Molecular Characterization of a Geranyl Diphosphate-Specific Prenyltransferase Catalyzing Stilbenoid Prenylation from Morus alba

Zhong Z, Zhu W, Liu S, Guan Q, Chen X, Huang W, et al.

Plant Cell Physiol, 2018 Nov 01;59(11):2214-2227.
PMID: 30020500 DOI: 10.1093/pcp/pcy138

Pharmaceutically active compounds from medical plants are attractive as a major source for new drug development. Prenylated stilbenoids with increased lipophilicity are valuable secondary metabolites which possess a wide range of biological activities. So far, many prenylated stilbenoids have been isolated from Morus alba but the enzyme responsible for the crucial prenyl modification remains unknown. In the present study, a stilbenoid-specific prenyltransferase (PT), termed Morus alba oxyresveratrol geranyltransferase (MaOGT), was identified and functionally characterized in vitro. MaOGT recognized oxyresveratrol and geranyl diphosphate (GPP) as natural substrates, and catalyzed oxyresveratrol prenylation. Our results indicated that MaOGT shared common features with other aromatic PTs, e.g. multiple transmembrane regions, conserved functional domains and targeting to plant plastids. This distinct PT represents the first stilbenoid-specific PT accepting GPP as a natural prenyl donor, and could help identify additional functionally varied PTs in moraceous plants. Furthermore, MaOGT might be applied for high-efficiency and large-scale prenylation of oxyresveratrol to produce bioactive compounds for potential therapeutic applications.

Matched MeSH terms: Organisms, Genetically Modified

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