Wachstum verschiedener Haematococcus pluvialis Stämme in unterschiedlichen Nährmedien und Entwicklung einer neuen Analyse zur Bestimmung des Astaxanthin-Gehaltes

Die weltweit verbreitete einzellige Grunalge Haematococcus pluvialis (Blutregenalge) bildet unter Stressbedingungen das Carotin Astaxanthin. Kommerziell wird Astaxanthin als Farbstoff in der Fischzucht, in der Kosmetikindustrie und als Nahrungserganzungsstoff verwendet. Fur eine moglichst effiziente Produktion ist es wichtig, optimale Wachstumsbedingungen zu schaffen und den besten Erntezeitpunkt mit dem grosstmoglichen Astaxanthin-Gehalt zu erkennen. Deswegen wurde in dieser Arbeit die Eignung vom Nahrmedium OHM (optimales Haematococcus Medium) fur die Kultivierung verschiedener H. pluvialis Stamme untersucht. Ausserdem wurde mittels einer Bildanalyse der Astaxanthin- Gehalt geschatzt und mit einer UV/VIS-Analyse verglichen werden. Dazu wurden vier H. pluvialis- Stamme (SAG 192.80, SAG 34-1B, CCAP 34/6, CCALA 793) wahrend 31 Tagen in den Nahrmedien OHM und in 3N-BBM+V kultiviert. Dabei konnte bestatigt werden, dass fur die Kultivierung von H. pluvialis das Nahrmedium OHM besser geeignet ist (dreimal besseres Wachstum) als 3N-BBM+V. Ausserdem zeigte sich, dass die Wahl des Stammes keinen Einfluss auf die maximale Zellzahl, die Wachstumsrate und die Verdoppelungszeit hat. Ebenfalls konnte gezeigt werden, dass sich der pH im OHM konstanter als im 3N-BBM+V verhalt. Bei den replizierten Kulturen des Stammes CCALA 793, die im OHM-Medium kultiviert wurden, wurde die Stressphase mittels nahrstoffarmen Mediums (ADaM) eingeleitet. Die Grossenverteilung der Zellen in der Kultur veranderte sich im Verlauf der Stressphase. Es kam zu einer Verschiebung der Zell-Durchmesser von kleinen Zellen zu grossen Zellen. Mit dem Programm Fiji konnte der Astaxanthin-Gehalt in Flachenprozent geschatzt werden. Anhand des Vergleichs mit der UV/VIS-Standardkurve konnte der genaue Astaxanthin-Gehalt in mg/g ermittelt werden. Der Rot-Anteil in Flachenprozent war ein guter Vorhersager fur den Astaxanthin-Gehalt von H. pluvialis-Zellen. Der lineare Zusammenhang konnte mit der folgenden Gleichung beschrieben werden: Astaxanthin-Gehalt [mg/g] = 5.9854 + 19.2006 × Rot-Anteil [Flachen-%].The worldwide common single-celled green alga Haematococcus pluvialis produces the carotene astaxanthin under stressful conditions. Commercially, astaxanthin is used as a dye in fish farming, in the cosmetics industry and as a dietary supplement. For an efficient production, it is important to achieve optimal growth conditions and to record the best harvest time with the highest possible astaxanthin content. Therefore, the suitability of the nutrient medium OHM (optimal Haematococcus medium) for the cultivation of various H. pluvialis strains was investigated. In addition, the astaxanthin content was to be estimated by an image analysis and compared with an HPLC analysis. For this purpose, four H. pluvialis strains (SAG 192.80, SAG 34-1B, CCAP 34/6, CCALA 793) were cultivated for 31 days in the nutrient mediums OHM and in 3N-BBM+V. In summary, the culture medium OHM was more suitable (three times better growth) for the cultivation of H. pluvialis than 3N-BBM+V. It is also shown that the choice of the strain had no significant influence on the maximum cell count, the growth rate and the doubling time. Furthermore, the pH behaved more consistently in the OHM than in the 3N-BBM+V. In the replicated cultures of the CCALA 793 strain, which are cultivated in the OHM medium, the stress phase was initiated by a nutrient-poor medium (ADaM). The size distribution of cells in the culture changed during the stress phase. There was a shift of the cell diameters from small cells to large cells. With the open source image processing program Fiji, the astaxanthin content could be estimated as a percentage by area. Based on the comparison with the UV/VIS standard curve, the exact astaxanthin content in mg/g could be determined. The red content given as area percent was a good predictor for the astaxanthin content of H. pluvialis cells. The linear relationship could be described with the following equation: astaxanthin content [mg/g] = 5.9854 + 19.2006 × red content [area-%]

Nitrous oxide emissions during microalgae cultivation

This thesis investigated the effect of nitrogen source and strain selection on nitrous oxide (N2O) emissions from Chlorella vulgaris cultures. Furthermore, nitrous oxide quantification using Fourier transform infrared spectroscopy (FTIR). This method was successful and suitable. It was found that light had a clear effect: Nitrous oxide production of Chlorella vulgaris was significantly increased when algae were incubated in the dark (1’300 nmol gDW-1 h-1) and supplied with nitrite as a nitrogen source compared to the cultures in light (130 nmol gDW-1 h-1). Moreover, the form of the nitrogen source plays a role: Chlorella vulgaris emitted negligible nitrous oxide when supplied with urea or ammonium as nitrogen sources (<18 nmol gDW-1 h-1). However, with nitrate as nitrogen source, the nitrous oxide emissions were around 205 nmol gDW-1 h-1. The largest emissions showed the cultures with nitrite as nitrogen sources and were about 1’314 nmol gDW-1 h-1. Despite considerable differences of nitrous oxide emissions depending on the Chlorella vulgaris strain used, these were statistically not significant. Strains SAG 9.88 and CCALA 924 emitted 2.5 to 5 times more nitrous oxide than the others.In dieser Arbeit wurden die Auswirkung der Stickstoffquelle und die Auswahl des Algenstammes auf die Lachgasemissionen (N2O) von Chlorella vulgaris-Kulturen untersucht. Ausserdem war der Nachweis von Lachgas mittels Fourier- Transformations-Infrarot-Spektroskopie (FTIR) erfolgreich und geeignet. Es wurde festgestellt, dass Licht einen klaren Einfluss hat: Die Lachgasproduktion von Chlorella vulgaris war deutlich erhöht, wenn die Algen im Dunkeln (1’300 nmol gDW-1 h-1) kultiviert wurden als bei den beleuchteten Kulturen (130 nmol gDW-1 h-1). Bei beiden Kulturen wurde Nitrit als Stickstoffquelle verwendet. Ausserdem wurde herausgefunden, dass die Form des Stickstoffdüngers eine Rolle spielt: Chlorella vulgaris emittierte vernachlässigbar wenig Lachgas, wenn Harnstoff oder Ammonium als Stickstoffquelle verwendet wurden (<18 nmol gDW-1 h-1). Mit Nitrat als Stickstoffquelle lagen die Lachgasemissionen jedoch bei 205 nmol gDW-1 h-1. Die größten Emissionen wiesen die mit Nitrit gedüngten Kulturen auf und lagen bei rund 1'314 nmol gDW-1 h-1. Trotz der gemessenen Unterschiede bei den Lachgasemissionen von verschiedenen Chlorella vulgaris Stämmen, waren diese statistisch nicht signifikant. Die Stämme SAG 9.88 und CCALA 924 emittierten 2.5 bis 5-mal mehr Lachgas als die anderen

Latent infection of human bocavirus accompanied by flare of chronic cough, fatigue and episodes of viral replication in an immunocompetent adult patient, cologne, Germany

Publisher Copyright: © 2016 The Authors.Introduction: The human bocavirus (HBoV) is a parvovirus and is associated with mild to lifethreatening acute or persisting respiratory infections, frequently accompanied by further pathogens. So far, there is limited knowledge on the mechanisms of persistence, and no reports on chronic infections or latency have been published so far. Case presentation: An immunocompetent male patient suffers from a chronic HBoV1 infection, i.e. viral DNA was detected in both serum and bronchoalveolar lavage (BAL) for >5 months without co-infections and with respiratory symptoms resolved spontaneously while receiving symptomatic treatment with montelukast and corticosteroids. Following the symptomatic medication of a chronic infection with HBoV1 viraemia indicating active viral replication lasting over 5 months, the patient cleared the viraemia and no further viral DNA was detectable in the BAL. However, by fluorescence in situ hybridization analyses of mucosal biopsies, it was shown that the virus genome still persisted in the absence of viral shedding but in a more compact manner possibly representing a supercoiled episomal form of this otherwise linear singlestranded DNA genome. This indicated the entry into a latency phase. Moreover, the cytokine profile and the IP-10/TARC ratio, a marker for fibrotization, seem to have been altered by HBoV1 replication. Although specific IgG antibodies were detectable during the whole observation period, they showed an apparently insufficient neutralising activity. Conclusion: On the one hand, these findings suggest that the symptomatic medication may have led to clearance of the virus from blood and airways and, moreover, that the viral DNA persists in the tissue as an altered episomal form favoured by lacking neutralising antibodies. This appears to be important in order to reduce possible long-term effects such as lung fibrosis.publishersversionPeer reviewe

Accounting for local temperature effect substantially alters afforestation patterns

Human intervention in forested ecosystems is hoped to perform a fundamental shift within the next decade by reverting current forest loss—a major source of CO2 emissions—to net forest gain taking up carbon and thus aiding climate change mitigation. The demanded extensive establishment of forests will change the local surface energy fluxes, and with it the local climate, in addition to competing with food and fiber production for land and water. Scenario building models encompass this competition for resources but have turned a blind eye to the biogeophysical (BGP) local surface energy flux disturbance so far. We combine the benefit of CO2 sequestration of afforestation/reforestation (A/R) with the additional incentive or penalty of local BGP induced cooling or warming by translating the local BGP induced temperature change to a CO2 equivalent. We then include this new aspect in the land-use model Model for Agricultural Production and their Impact on the Environment (MAgPIE) via modifying the application of the price on greenhouse gases (GHGs). This enables us to use MAgPIE to produce A/R scenarios that are optimized for both their potential CO2 sequestration and the CO2 equivalent local BGP effect, as well as the socio-economic trade-offs of A/R. Here we show that optimal A/R patterns are substantially altered by taking the local BGP effects into account. Considering local cooling benefits of establishing forests triples (+203.4%) the viable global A/R area in 2100 from 116 to 351 Mha under the conditions of the shared socioeconomic pathway 2 (SSP2) scenario driven by the same GHG price. Three quarters (76.0%, +179 Mha) of the additionally forested area is established in tropical climates alone. Therefore, a further neglect of BGP effects in scenario building models undervalues the benefit of tropical forests while simultaneously running the risk of proposing counterproductive measures at high latitudes. However, the induced focus on tropical forestation intensifies the competition with food production where forests contribute most to mitigation. A/R related trade-offs need to be considered alongside their climate benefit to inhibit unintentional harm of mitigation efforts.AXISERANETFederal Ministry of Education and Research (BMBF)German Aerospace Center (DLR)European UnionPeer Reviewe

Type I and type II interferon responses in two human liver cell lines (Huh-7 and HuH6)

AbstractMost studies investigating the biology of Hepatitis C virus (HCV) have used the human hepatoma cell line Huh-7 or subclones thereof, as these are the most permissive cell lines for HCV infection and replication. Other cell lines also support replication of HCV, most notably the human hepatoblastoma cell line HuH6. HCV replication in cell culture is generally highly sensitive to interferons (IFNs) and differences in the IFN-mediated inhibition of virus replication may reflect alterations in the IFN-induced antiviral response inherent to different host cells. For example, HCV replication is highly sensitive to IFN-γ treatment in Huh-7, but not in HuH6 cells. In this study, we used microarray-based gene expression profiling to compare the response of Huh-7 and HuH6 cells to stimulation with IFN-α and IFN-γ. Furthermore, we determined whether the resistance of HCV replication in HuH6 cells can be linked to differences in the expression profile of IFN-regulated genes. Although both cells lines responded to IFNs with rapid changes in gene expression, thereby demonstrating functional type I and type II signaling pathways, differences were observed for a number of genes. Raw and normalized expression data have been deposited in GEO under accession number GSE68927

Genotype-by-Diet Interactions for Larval Performance and Body Composition Traits in the Black Soldier Fly, Hermetia illucens

Further advancing black soldier fly (BSF) farming for waste valorisation and more sustainable global protein supplies critically depends on targeted exploitation of genotype-phenotype associations in this insect, comparable to conventional livestock. This study used a fully crossed factorial design of rearing larvae of four genetically distinct BSF strains (FST: 0.11–0.35) on three nutritionally different diets (poultry feed, food waste, poultry manure) to investigate genotype-by-environment interactions. Phenotypic responses included larval growth dynamics over time, weight at harvest, mortality, biomass production with respective contents of ash, fat, and protein, including amino acid profiles, as well as bioconversion and nitrogen efficiency, reduction of dry matter and relevant fibre fractions, and dry matter loss (emissions). Virtually all larval performance and body composition traits were substantially influenced by diet but also characterised by ample BSF genetic variation and, most importantly, by pronounced interaction effects between the two. Across evaluated phenotypes, variable diet-dependent rankings and the lack of generally superior BSF strains indicate the involvement of trade-offs between traits, as their relationships may even change signs. Conflicting resource allocation in light of overall BSF fitness suggests anticipated breeding programs will require complex and differential selection strategies to account for pinpointed trait maximisation versus multi-purpose resilience

From Refugees to Workers: Mapping Labour-Market Integration Support Measures for Asylum Seekers and Refugees in EU Member States - Volume II: Literature Review and Country Case Studies

This study intends to provide a better understanding of the challenges with regard to the integration of refugees into the labour-markets. What are the strategies and practices implemented in different EU Member States to facilitate access to employment? What do we know about their effectiveness? What are good practices and lessons learned in different countries? The study includes detailed case studies for the following nine EU Member States: Austria, Denmark, France, Germany, Italy, the Netherlands, Spain, Sweden and the United Kingdom. The research points to the need for increased coordination at all levels, the conditions for successful public-private partnerships, and the adequate sequence of work integration and language learning, for example. Not least, it makes clear that finding effective ways to bring refugees to work will prove key for Europe's future.It has been produced by the Migration Policy Centre (MPC) at the European University Institute in Florence.Search also for: Volume I of the report "Comparative Analysis and Policy Findings"

Overcoming global inequality is critical for land-based mitigation in line with the Paris Agreement

Transformation pathways for the land sector in line with the Paris Agreement depend on the assumption of globally implemented greenhouse gas (GHG) emission pricing, and in some cases also on inclusive socio-economic development and sustainable land-use practices. In such pathways, the majority of GHG emission reductions in the land system is expected to come from low- and middle-income countries, which currently account for a large share of emissions from agriculture, forestry and other land use (AFOLU). However, in low- and middle-income countries the economic, financial and institutional barriers for such transformative changes are high. Here, we show that if sustainable development in the land sector remained highly unequal and limited to high-income countries only, global AFOLU emissions would remain substantial throughout the 21st century. Our model-based projections highlight that overcoming global inequality is critical for land-based mitigation in line with the Paris Agreement. While also a scenario purely based on either global GHG emission pricing or on inclusive socio-economic development would achieve the stringent emissions reductions required, only the latter ensures major co-benefits for other Sustainable Development Goals, especially in low- and middle-income regions