Limits of Concern: suggestions for the operationalisation of a concept to determine the relevance of adverse effects in the ERA of GMOs

Background

The European Food Safety Authority proposed a concept for the environmental risk assessment of genetically modified plants in the EU that is based on the definition of thresholds for the acceptability of potential adverse effects on the environment. This concept, called Limits of Concern (LoC), needs to be further refined to be implemented in the environmental risk assessment of genetically modified organisms.

Methods

We analyse and discuss how LoC can be defined for the environmental risk assessment for three different types of genetically modified plants. We outline protection goals relevant to the genetically modified plants in question and discuss existing concepts and suggestions for acceptability thresholds from the environmental risk assessment of different regulatory areas. We make specific recommendations for the setting and use of LoC for each type of genetically modified plant.

Results

The LoC concept can be suitably applied for the environmental risk assessment of genetically modified organisms, if the different protection goals in agro-environments are specifically considered. Not only biodiversity protection goals but also agricultural protection goals need to be addressed. The different ecosystem services provided by weeds inside and outside agricultural fields have to be considered for genetically modified herbicide-tolerant crops. Exposure-based LoCs are suggested based on knowledge about dose–effect relationships between maize pollen and non-target Lepidoptera for insect-resistant maize. Due to the long-term nature of biological processes such as spread and establishment, LoCs for genetically modified oilseed rape should be defined for the presence of the genetically modified plant or its genetically modified traits in relevant protection goals.

Conclusions

When setting LoCs, the focus should be on protection goals which are possibly affected. Potential overlaps of the LoC concept with the ecosystem service concept have to be clarified to harmonise protection levels in the agro-environment for different stressors. If additional impacts on agro-biodiversity resulting from the cultivation of genetically modified plants are to be avoided, then high protection levels and low thresholds for acceptable effects (i.e. LoC) should be set.

     

European Court of Justice ruling regarding new genetic engineering methods scientifically justified: a commentary on the biased reporting about the recent ruling

In July 2018, the European Court of Justice (Case C-528/16) ruled that organisms obtained by directed mutagenesis techniques are to be regarded as genetically modified organisms (GMOs) within the meaning of Directive 2001/18. The ruling marked the next round of the dispute around agricultural genetic engineering in Europe. Many of the pros and cons presented in this dispute are familiar from the debate around the first generation of genetic engineering techniques. The current wave of enthusiasm for the new genetic engineering methods, with its claim to make good on the failed promises of the previous wave, seems to point more to an admission of failure of the last generation of genetic engineering than to a true change of paradigm. Regulation is being portrayed as a ban on research and use, which is factually incorrect, and the judges of the European Court of Justice are being defamed as espousing “pseudoscience”. Furthermore, this highly polarised position dominates the media reporting of the new techniques and the court’s ruling. Advocates of the new genetic engineering techniques appear to believe that their benefits are so clear that furnishing reliable scientific evidence is unnecessary. Meanwhile, critics who believe that the institution of science is in a serious crisis are on the increase not just due to the cases of obvious documented scientific misconduct by companies and scientists, but also due to the approach of dividing the world into those categorically for or against genetic engineering. In this construct of irreconcilable opposites, differentiations fall by the wayside. This article is a response to this one-sided and biased reporting, which often has the appearance of spin and lacks journalistic ethics that require journalists to report on different positions in a balanced and factual manner instead of taking positions and becoming undeclared advocates themselves.     

Fast and complete removal of the 5-fluorouracil drug from water by electro-Fenton oxidation

Cytostatic drugs are a troublesome class of emerging pollutants in water owing to their potential effects on DNA. Here we studied the removal of 5-fluorouracil from water using the electro-Fenton process. Galvanostatic electrolyses were performed with an undivided laboratory-scale cell equipped with a boron-doped diamond anode and a carbon felt cathode. Results show that the fastest degradation and almost complete mineralization was obtained at a Fe²⁺ catalyst concentration of 0.2 mM. The absolute rate constant for oxidation of 5-fluorouracil by hydroxyl radicals was 1.52 × 10⁹ M^?1 s^?1. Oxalic and acetic acids were initially formed as main short-chain aliphatic by-products, then were completely degraded. After 6 h the final solution mainly contained inorganic ions (NH₄ ⁺, NO₃ ^? and F^?) and less than 10% of residual organic carbon. Hence, electro-Fenton constitutes an interesting alternative to degrade biorefractory drugs.     

Nanosystems for drug delivery of coenzyme Q10

Coenzyme Q10 is an antioxidant present in the human body. Coenzyme Q10 has an essential role in various biochemical reactions. The deficiency of coenzyme Q10 in the body leads to disorders including neurological degeneration, ageing and cancer. Clinical trials have tested coenzyme Q10 as a drug or a dietary supplement. However, the major pharmaceutical issue of coenzyme Q10 delivery is its high molecular weight and poor water solubility. This limitation leads to its poor oral bioavailability. Several methods have been designed to overcome the poor water solubility of coenzyme Q10, such as size reduction and ionization. This article presents nanotechnology-based drug delivery systems for coenzyme Q10 with special emphasis on pharmacokinetic perspectives and clinical relevance. Systems include nanoparticles, solid dispersions, liposomes, nanoemulsions, self-emulsifying drug delivery systems, nanostructured lipid carriers, cyclodextrins and nanocapsules.     

Nanoscience in food and agriculture: research,industries and patents

High population rise and climate changes are increasing issues of agricultural production and food safety. Nanotechnology is finding revolutionary applications to improve agricultural and food systems, notably for better crop production and food preservation. Here we review research, industrial and patent trends of nanoscience in food and agriculture. In a literature survey, we found 44.6% publications in the nano-food research area during the years 2013–2015 and 59.09% publications in the nano-agriculture research area during 2012–2015. USA is leading in the development of nanotechnology firms with a maximum share of 75.5% of the total firms, followed by Germany and France with 8.10 and 4.74%, respectively. USA is leading in the nano-food research with 22 granted patents, whereas China is placed first in nano-agriculture research with 28 granted patents during assessment years 2011–2015. Nano-food research focused mainly on nano-food packaging with 76.84% contributions, whereas in nano-agriculture research, focus has been on nano-fertilizers with 90% contributions. Germany, France, Korea, Italy, Czech Republic, Slovenia and Slovak republic have more than 20% of dedicated nanotechnology firms. A growth of about 45% in nano-food patents has been observed for USA during 2011–2015, and China is leading in the nano-agriculture patents with an increase of 60.7% during 2012–2015.     

High zinc removal from water and soil using struvite-supported diatomite obtained by nitrogen and phosphate recovery from wastewater

Zinc is known as an essential element of human life. However, excessive zinc discharge into water and soil causes water pollution, leading to serious health issues such as septicemia, meningitis and iron-deficiency anemia. Here, a novel material made of struvite-supported diatomite was obtained from eutrophic water treated by mesoporous MgO-modified diatomite. This material was applied for zinc remediation in aqueous solutions and contaminated soils to test the reuse of P-containing products. Struvite-supported diatomite was characterized by field emission scanning electron microscopy and X-ray diffraction. Results show that the maximum removal efficiency of Zn(II) from wastewater streams reached 90.54% at an initial pH of 5 and struvite-supported diatomite dosage of 0.3 g/L. Moreover, the X-ray diffraction patterns of precipitates after Zn(II) sorption show that the combination between zinc and the phosphate group played a key role for zinc removal in solution. For Zn-contaminated soils amended with 10% struvite-supported diatomite, available Zn decreased by 65.38% and acid soluble Zn decreased by 56.9% after 56 days.     

High CO<Subscript>2</Subscript> adsorption by amino-modified bio-spherical cellulose nanofibres aerogels

Climate change has become increasingly serious due to the greenhouse effect. It is therefore necessary to control the content of greenhouse gases such as carbon dioxide in the atmosphere, using, for instance, CO₂-adsorbing materials. Here, we synthesized ultra-lightweight and spherical cellulose nanofibres aerogels by a suspension titration method using an efficient amination process. These functional materials with high porosity, higher than 96.54%, and three-dimensional network structure, were prepared by freeze-drying spherical cellulose nanofibres hydrogel. Their maximum CO₂ adsorption capacity reaches 1.78 mmol/g, and they show excellent regeneration, of more than 10 cycles. This synthesis of bioaerogels represents a new method for the preparation of bio-CO₂ adsorbents.     

Free radical-assisted electrocoagulation processes for wastewater treatment

The combination of electrocoagulation with another process is a promising approach to enhance the removal efficiency of water pollutants. For instance, free radical-assisted electrocoagulation is a new combination showing higher performance. There are different combinations depending on the free radical source. This article reviews free radical-assisted electrocoagulation processes. We discuss electrocoagulation mechanisms; ozone-assisted electrocoagulation processes; advanced oxidation-assisted electrocoagulation processes; and ultrasound-assisted electrocoagulation. We present kinetic models used in free radical-assisted electrocoagulation, scale-up of free radical-assisted electrocoagulation and cost estimation. The major points are: most of the available studies have been done at laboratory scale with synthetic wastewater, and lack holistic and systematic approaches to consider the process complexity. The performance of the combined process is improved, and the removal efficiency is increased especially with ozone-assisted electrocoagulation, which gives a removal efficiency of more than 95%. The use of ultrasound energy with electrocoagulation is advantageous in reducing the problem of electrode passivation.     

Effect of herbicides on glutathione S-transferases in the earthworm, <Emphasis Type="Italic">Eisenia fetida</Emphasis>

AIM AND BACKGROUND: Earthworms have been studied as a readily available, easily maintainable and cheap test species for assessing chemical pollution, and may be an alternative to in vivo rodent bioassays. The current investigation aims to characterize detoxification enzymes in Eisenia fetida and stress response against two herbicides with different modes of action, namely, fenoxaprop and metolachlor. METHODS: Herbicides were applied to soil containing earthworms. Animals were then collected, sacrificed and shock-frozen. Extracted protein was analyzed for glutathione S-transferase (GST) activity using CDNB (1-chloro-2,4-dinitrobenzene), DCNB (1,2-dichloro-4-nitrobenzene), pNBC (p-nitrobenzylchloride), PNOBC (p-nitrobenz-o-ylchloride) and selected herbicides. GST isoenzymes were partially purified by affinity chromatography and molecular weights were estimated by SDS-PAGE. RESULTS: In E. fetida protein extracts, GST activity towards model compounds ranked as CDNB>DCNB>PNBOC>PNBC. Fluorodifen was not conjugated at all, but fenoxaprop and metolachlor were conjugated at low rates. Furthermore, the GST isoenzyme pattern changed during the incubation with herbicides, either due to stress or as a defense reaction. After incubation with monochlorobimane, a strong fluorescence of the intestinal tract and the intersegments was observed, indicating organ-specific GST induction. DISCUSSION: According to the author's knowledge, here, for the first time, evidence is presented that E. fetida GST are also capable of conjugating a wider range of xenobiotic substrates. Different forms of GST were observed and changes in GST isoforms due to the herbicide treatment were also noticed. GST conjugation rates varied between different herbicides used in this experiment. It might be assumed that herbicides may well be detoxified by earthworms, to a certain extent, but that they are also potent stress factors influencing the detoxification system of the animal. High doses or long exposure might lead to deleterious effects on earthworms and limit their survival rate. The use of the animals as bioindicators for herbicides and herbicide residues seems very promising, but is surely influenced by the lack of detoxification for some compounds. CONCLUSIONS: Conjugation of several xenobiotics with model substances and herbicides is proven in the earthworm E. fetida. However, E. fetida has only limited capabilities of detoxifying herbicidal compounds. Different isoforms of GST were involved and altered in their activity after treatment. RECOMMENDATIONS AND PERSPECTIVES: The accumulation of GS-conjugates and their determination via fluorescence microscopy is a quick and secure, additional marker for exposure that should be further developed to complement existing biotests. The described methods and endpoints might help to understand the complex reaction of earthworms towards herbicides and lead to an adapted test methodology.