10 research outputs found

    Parasite responses to pollution: what we know and where we go in ‘Environmental Parasitology’

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    Evaluation of an improved RNA/DNA quantification method in a common carp (Cyprinus carpioLinnaeus 1758) larval feeding trial withArtemia, two nematodes (Panagrellus redivivusLinnaeus 1758,Panagrolaimussp. Fuchs 1930) and dry feed

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    The RNA/DNA ratio commonly used as proxy for the nutritional condition of fish larvae is affected by RNA degradation during analysis. For evaluation of two strategies to improve RNA integrity, a three-week feeding trial was carried out to assess the suitability of two nematode species (fam. Panagrolaimidae) as feed for newly hatched carp larvae (Cyprinus carpio) in comparison to Artemia nauplii (Artemia sp.) and a commercial dry feed. Aiming for an increased reproducibility of RNA/DNA determination, a high-salt inactivation (RNA later) as well as a targeted approach with a recombinant RNase inhibitor were compared to the classical protocol using lab chip technology. Improved RNA integrity was observed with high-salt inactivation when compared with a strategy applying a specific RNase inhibitor or the classic protocol. Carp larvae fed Artemia for 2 weeks and then dry feed for 1 week revealed the best overall growth performance as well as survival [83.0 ± 35.2 mg fresh weight (FW), 20.0 ± 2.4 mm total length (TL), 86.6 ± 11.7% survival]. Larvae fed the nematode species Panagrellus redivivus for 1 week and subsequently dry feed for 2 weeks (37.4 ± 29.1 mg FW, 14.7 ± 2.8 mm TL, 76.0 ± 6.0% survival) performed better than larvae fed with dry feed alone (28.2 ± 29.6 mg FW, 14.3 ± 2.9 mm TL, 54.3 ± 14.2% survival) or those receiving Panagrellus for 2 weeks. Between both nematode species, Panagrellus was a better feed with regard to growth performance and survival. RNA/DNA ratios ranged between 0.65 ± 0.27 (8 days post-hatch) and 1.96 ± 0.63 (22 days post-hatch) and were in the same treatment order as the other growth parameters. RNA/DNA ratios were significantly correlated with the growth rate, and decreasing RNA/DNA ratios in larger larvae may reflect decreasing growth rates with size rather than decreased nutritional status. Here, an improved RNA/DNA ratio protocol is presented in a feeding trial that reveals the suitability of nematodes as a first feed for common carp larvae

    Polypropylene numerical photoageing simulation by dose - response functions with respect to irradiation and temperature: ViPQuali project

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    The aim of the joint project ViPQuali (Virtual Product Qualification) was to describe a component’s ageing behaviour in a given environment, by numerical simulation. Having chosen polypropylene (PP) as the material, which does not show sensitivity to moisture, the relevant weathering parameters of the dose–response functions could be limited to spectral irradiance and temperature. In artificial irradiation tests, for PP plates of varied stabiliser content, spectral sensitivity as well as temperature dependence of irradiation-caused crack formation was quantified. For that purpose, samples were exposed both to artificial weathering tests at various constant temperatures and to spectrally resolved irradiation. The temperature dependence could be modelled by an Arrhenius fit. For fitting the spectral sensitivity, a plateau function was chosen. Subsequently, the stabiliser content was parameterised and extrapolated. The formed dose–response functions were incorporated into a Computational Fluid Dynamics (CFD) software program, simulating the environment of a sample within a Phoenix-exposed IP/DP (Instrument Panel/Door Panel box) box, based on sun position and weather conditions, including radiation interactions. Observed local effects as well as the general ageing advance of PP hats are compared with respect to simulation and experiment. Resulting from this project, for this most simple example of PP of varied stabiliser content, the time to failure can be estimated for each weathering exposure environment with known time-resolved irradiance and temperature conditions

    A Rapid CRISPR/Cas-based Mutagenesis Assay in Zebrafish for Identification of Genes Involved in Thyroid Morphogenesis and Function

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    The foregut endoderm gives rise to several organs including liver, pancreas, lung and thyroid with important roles in human physiology. Understanding which genes and signalling pathways regulate their development is crucial for understanding developmental disorders as well as diseases in adulthood. We exploited unique advantages of the zebrafish model to develop a rapid and scalable CRISPR/Cas-based mutagenesis strategy aiming at the identification of genes involved in morphogenesis and function of the thyroid. Core elements of the mutagenesis assay comprise bi-allelic gene invalidation in somatic mutants, a non-invasive monitoring of thyroid development in live transgenic fish, complementary analyses of thyroid function in fixed specimens and quantitative analyses of mutagenesis efficiency by Illumina sequencing of individual fish. We successfully validated our mutagenesis-phenotyping strategy in experiments targeting genes with known functions in early thyroid morphogenesis (pax2a, nkx2.4b) and thyroid functional differentiation (duox, duoxa, tshr). We also demonstrate that duox and duoxa crispants phenocopy thyroid phenotypes previously observed in human patients with bi-allelic DUOX2 and DUOXA2 mutations. The proposed combination of efficient mutagenesis protocols, rapid non-invasive phenotyping and sensitive genotyping holds great potential to systematically characterize the function of larger candidate gene panels during thyroid development and is applicable to other organs and tissues.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

    Principles of sound ecotoxicology

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    We have become progressively more concerned about the quality of some published ecotoxicology research. Others have also expressed concern. It is not uncommon for basic, but extremely important, factors to apparently be ignored. For example, exposure concentrations in laboratory experiments are sometimes not measured, and hence there is no evidence that the test organisms were actually exposed to the test substance, let alone at the stated concentrations. To try to improve the quality of ecotoxicology research, we suggest twelve basic principles that should be considered, not when presenting findings to the regulators, but at the stage of experimental design. These principles range from accurately defining the exposure through to carefully considering essential aspects of experimental design as well as unbiased analysis and reporting of the results. Although not all principles will apply to all studies, we offer these principles in the hope that they will improve the quality of the science that is presented to regulators. Science is an evidence-based discipline, and it is important that we and the regulators can trust the evidence presented to us. Significant resources often have to be devoted to refuting the results of poor research when those resources could be utilised more effectively
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