4 Material und Umwelt
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Eingeladener Vortrag
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The Cairo Genizah is a well-known “archive” discovered in the 19th century in Ben Ezra Synagogue in Fustat, a district in Old Cairo (Egypt), located south of the center of modern Cairo. The giant collection of mostly Jewish documents that vary in genres, languages and writing supports contains a large number of early medieval Hebrew manuscripts, mostly in fragmentary form. The larger part of the Cairo Genizah is stored today in the Cambridge University Library, containing about 200,000 fragments.
The Genizah provides sources for the literary, linguistic, historical studies of the various aspects Jewish and non-Jewish life. As the documents attest, at least two Jewish communities co-existed in Fustat up to 11th century: a so-called Babylonian and Palestinian. These communities had different leaders, different traditions and lived independently. The differences manifest themselves also in the paleographical properties of the manuscripts produced by each community.
Although Genizah manuscripts have attracted a great deal of scholarly attention, the material aspects of these documents seem to have been largely neglected. The current study aims to obtain information on the composition of the writing supports and the inks used, in order to obtain insights on trade, local production techniques and social structures.
Can we extend the differences detected by the paleographical analysis to differences in composition of the writing materials ? Can we relate the differences in the composition to a community, to the manuscript type (legal, private...), or to another criterion?
The Cairo Geniza is an “archive” discovered in the 19th century in Ben Ezra Synagogue in Fustat, a district in Old Cairo (Egypt). The collection of mostly Jewish documents that vary in genres, languages and writing supports contains a large number of early medieval Hebrew manuscripts.
The Geniza provides sources for the literary, linguistic, historical studies of the various aspects Jewish life. As the documents attest, at least two Jewish communities co-existed in Fustat up to 11th century: a so-called Babylonian and Palestinian. These communities had different leaders, different traditions and lived independently. The differences seem to manifest themselves also in the paleographical, codicological and some material properties of the manuscripts produced by each community.
The aim of this project is to compare the inks used in the Jewish documents depending on different variables: support (paper, parchment), purpose of the manuscript (legal, private, religious), provenance of the scribe. For the determination of the inks type and composition we had to choose non-invasive, non-destructive and portable techniques to analyze the corpus directly on-situ.
The analyses were carried out with a micro-X-ray spectrometer ArtTAX® (Bruker GmbH) and a digital stereomicroscope features built-in LED illumination at 395 nm and 940nm, Dino Lite.
We would like to stress that though the methods of material analysis listed above have been successfully employed in the field of cultural heritage and conservation including ancient and medieval manuscripts they have not yet been used to study fragments from the Cairo Genizah. Therefore, we believe that this research project is a pioneering study that will provide new insights into the history of Hebrew writing materials, their production techniques and materials and, thus, contribute new data to the field of Hebrew paleography.
The quest for the mixed inks
(2018)
In this article, we would like to share our observations concerning the inks produced by intentionally mixing soot or charcoal with tannin extracts or iron-gall ink. Aside from Zerdoun’s mention in her outstanding review of written sources, “Les encres noires au Moyen-Âge”, this ink category has received little if any attention from scholars and scientists. And yet, if analytically attested, the use of such inks could serve as an additional category to classify and distinguish the writing inks on the historical socio-geographic map of the writing inks we are trying to build.
This application note presents how EnviroESCA can be used to analyze bacterial samples under near ambient pressure conditions in various states of hydration using different levels of humidity. Such investigations of bacterial cell wall surfaces in their hydrated state are essential for studying biological interfaces at work.
The use of innovative near-ambient pressure (NAP-)XPS instrumentation allows the detailed analysis of irregularly-surfaced biofilms. NAP-XPS enables the surface analysis of bacterial samples in their natural hydrated state without complex sample preparation techniques such as freeze-drying or fast-freezing, which are needed for XPS analysis in ultrahigh vacuum.
The first documentation of fuel biodeterioration dates back to the late 19th century. However, extensive studies concerning the microbial fuel contamination started in 1980’s. Polymeric fuel storage tanks containing diesel and biodiesel provide environmental conditions for microbial growth. Several studies demonstrated that bacteria, which were found in contaminated fuel systems, can use fuels as macronutrient; but such bacteria can also cause microbiologically influenced corrosion and fouling.
The aim of this study is to investigate the initial attachment behavior of bacteria, isolated from a diesel contamination, on neat and photooxidized high-density polyethylene (PE-HD). Two common PE-HD’s, less- and biodiesel-stabilized, were radiated to UV light representing a tank exposed to sunlight. The effect of photooxidiation on PE-HD’s surface were characterized chemically by Fourier-transform infrared spectroscopy (FTIR). The attached bacteria Pseudomonas aeruginosa and Bacillus subtilis on the polymer surface were evaluated by fluorescence microscopy and colony-forming unit tests (CFU).
There is a need for an assessment of the emission properties of volatile organic compounds (VOCs) from consumer products. A method comparison was carried out to evaluate adapted and cost-effective procedures for such items. Smaller and automated emission chambers de-picted similar kinetics compared to a 203 L standard chamber. Toy samples made of PVC (Polyvinyl chloride) emitted more VOCs compared to other tested polymeric products. The emissions from 2 selected samples were studied to allow an evaluation of the resulting room concentration and external exposure of a child. Obtained concentrations were not of concern.
There is a need for an assessment of the emission properties of volatile organic compounds (VOCs) from consumer products. A method comparison was carried out to evaluate adapted and cost-effective procedures for such items. Smaller and automated emission chambers depicted similar kinetics compared to a 203 L standard chamber. Toy samples made of PVC (Polyvinyl chloride) emitted more VOCs compared to other tested polymeric products. The emissions from 2 selected samples were studied to allow an evaluation of the resulting room concentration and external exposure of a child. Obtained concentrations were not of concern.
Advanced codicological studies of Cod. germ. 6 (Hamburg, Staats- und Universitätsbibliothek): Part 2
(2018)
The work presented here follows the article Combining Codicology and X-Ray Spectrometry to Unveil the History of Production of Codex germanicus 6 (Staats- und Universitätsbibliothek Hamburg), published in 2014.1 It confirms the main result of the previous article: the Artusnotiz, the fourth text in the bound manuscript, must have been introduced as the last one. This paper offers further details of the codex production, based on the composition of the black and red inks collected in four measurement campaigns. Furthermore, using imaging μ-XRF, we succeeded in understanding the strong variation of the composition of the red inks in the initials of all the texts except for Parzival and Jeanne d’Arc.
While studying the socio-geographic history of inks, division 4.5 of the BAM (Bundesanstalt für Materialforschung und Prüfung) together with the Centre for the Study of Manuscript Cultures in Hamburg has developed a protocol for ink analysis. It consists of a primary screening to determine the type of the ink and a subsequent in-depth analysis using several spectroscopic techniques: XRF, FTIR, and Raman. In most cases, we can obtain satisfactory results using a non-invasive protocol. However, mixed inks that contain no metals evade such a protocol. These inks constitute a heterogeneous group of media used especially in the Middle East and the Islamicate world since at least the 10th century; they are characterized by blending carbon ink and tannins, with or without the addition of vitriol.
Our own research aims primarily at recreating a socio-geographic history of inks, parchment, and papyrus and includes the comparative analysis of the writing materials of the Dead Sea Scrolls, ink and papyrus in Ancient and Hellenistic Egypt, and inks in documents from various contemporary medieval communities in Fustat (first nucleus of Cairo). During many years of study, we concluded that the continuous production of Coptic manuscripts from late Antiquity to the Middle Ages offers a unique opportunity for historical study of the ink in a large geographic area. Thanks to the collaboration with the ERC project “PAThs” (www.paths.uniroma1.it), based at the University of Rome La Sapienza, and within the activities of a PhD research dedicated to this topic, we therefore created a new branch of our project focused entirely on the analysis of Coptic inks, pigments, and dyes. This pioneering systematic study of writing materials coming from a specific area and time frame (5th-10th century) aims not only at a better understanding of the complex Coptic multicultural and plurilingual society, but also and mainly at clarifying the links among the Coptic and other societies between the ancient and medieval eras. Finally, it will cast light on the history of the technological development of inks in the eastern world, from Antiquity to the middle ages.
While studying the socio-geographic history of inks, division 4.5 of the BAM (Bundesanstalt für Materialforschung und Prüfung) together with the Centre for the Study of Manuscript Cultures in Hamburg has developed a protocol for ink analysis. It consists of a primary screening to determine the type of the ink, and a subsequent in-depth analysis using several spectroscopic techniques. The XRF elemental analysis aims primarily at understanding the fingerprints of inks containing metals, making it possible to distinguish among different inks.
Our own research aims primarily at recreating a socio-geographic history of inks, parchment, and papyrus and includes the comparative analysis of the writing materials of the Dead Sea Scrolls, ink and papyrus in Ancient and Hellenistic Egypt, and inks in documents from various contemporary medieval communities in Fustat (first nucleus of Cairo). During many years of study, we concluded that the continuous production of Coptic manuscripts from late Antiquity to the Middle Ages offers a unique opportunity for historical study of the ink in a large geographic area. Thanks to the collaboration with the ERC project “PAThs” (www.paths.uniroma1.it), based at the University of Rome La Sapienza, and within the activities of a PhD research dedicated to this topic, we therefore created a new branch of our project focused entirely on the analysis of Coptic inks, pigments, and dyes. This pioneering systematic study of writing materials coming from a specific area and time frame (5th-10th century) aims not only at a better understanding of the complex Coptic multicultural and plurilingual society, but also and mainly at clarifying the links among the Coptic and other societies between the ancient and medieval eras. Finally, it will cast light on the history of the technological development of inks in the eastern world, from Antiquity to the middle ages.
It is well documented that throughout Antiquity, ancient Egyptians used mostly carbon inks as a writing material. In Late Antiquity, some metals started to be added to carbon based inks. We have records of five manuscripts from the Dead Sea Scrolls collection whose carbon inks were found to contain copper Also, lead was recently found as an additive in carbon inks on a charred fragment from Herculaneum. Furthermore, the earliest evidence of iron-gall ink was found in the Book of Proverbs (Codex Ms. Berol. orient. oct. 987) dating to the third fourth centuries ce. It has been suggested that along with carbon and iron-gall inks, there is no reason to think that purely tannin inks were not also in use in Egypt. However, so far, we just have evidence of a copper-tannin ink identified in a number of documents from Egypt in the first third centuries bce. In an attempt to fill this gap in this extremely fragmented scenario during our studies of the socio-geographic history of inks, we arrived at the conclusion that the continuous production of Coptic manuscripts from Late Antiquity to the Middle Ages offers a unique opportunity for the historical study of inks across a large geographic area.
The manuscript Codex germanicus 1 (Cod. germ. 1) of the Staats- und Universitätsbibliothek Hamburg is a fifteenth-century German-language manuscript. It comprises two codicological units and has an especially complex developmental history. To trace this developmental history, neglected until now in the research literature, the manuscript was investigated, for the first time not solely with classical codicological and palaeographical methods, but also with the aid of X-ray fluorescence spectroscopy, in order to determine the composition of the writing materials. These methods made it possible, first, to support and check palaeographic findings and, second, to gain information about the stratigraphy of the manuscript where palaeographic methods find their limits – in regard to short entries, rubrications, and non-alphabetical signs.
The SCALE Project is a Horizon2020 Project (GA°730105) that aims to develope a secure supply chain for Scandium in Europe. To achieve that, the whole value chain is investigated and new methodologies and techniques are being developed. In BAM we are characterizing potential Scandium-bearing industrial by-products.
Small grain size fractions of municipal solid waste incineration (MSWI) bottom ash (BA) contain elemental and chemically bonded metals in appreciable amounts, especially copper. Investigations with a centrifugal concentrator were carried out using samples, which had been treated with different impact devices. Partly the impacts of single devices were visible in outlines, but mostly the surfaces of the metals were still covered with mineral residues. To generate copper rich fractions, parameters of the separation process should ensure mass yields <2%, best between 0.5 and 1%. The density of the concentrates is then generally >3.2 g/cm3. Best results were achieved with material from a special treatment train which is adapted to grain sizes <2 mm. Reliable copper contents between 20 and 40% (single runs >50%) could be realised.
However, considering the required mass yields, referred to this subset only 150–170 g copper would be recovered from 1 Mg MSWI BA. Furthermore, this copper is only in parts present in elemental form. A
dependable added value of such treatment step seems to be given only, if that enables a recycling of the mineral aggregates (tailings) in parallel to avoid landfill costs.
Quality Improvement of VOC chamber emission tests observed for 10 years with round robin tests
(2018)
In Europe, the Construction Products Regulation (CPR, 2011/305/EU) sets basic requirements (BR) on how construction works must be designed and built. BR 3 “hygiene, health and the environment” states low emissions of toxic gases, Volatile Organic Compounds (VOCVOCs), particles, etc. from building materials. Meanwhile, a worldwide network of professional Commercial and non-commercial laboratories performing emission tests for the evaluation of products for interior use has been established.
Therefore, comparability of test results must be ensured. The participation in Round Robin Tests (RRTRRTs) is a means to prove a laboratory’s proficiency. Since 2006 BAM offers such proficiency test for emission chamber test every two years.
Any commercially available product on the market can be used as reference material, provided it is thoroughly characterised. However, these materials often emit only a few and material specific VOCs and appropriate homogeneity is not given. Therefore, alternative materials should be found. BAM used different materials over the years. Currently, the best suited reference material is a cured lacquer surface which contained several VOCs added to the liquid lacquer system. The round robin tests 2014, 2016 and 2018 were conducted with such a system.
Compounds like Styrene or n-Alkanes like Decane or Tridecane were tested in many of the proficiency test operated by BAM. Up to now the relative standard deviation of these compounds decrease continuously. Starting with values about 30 % in 2008 we reached values for the mentioned compounds 15 to 20 % now. But even for more polar compounds improved results were received.
Over the years the results of a large group of laboratories detected many compounds with much higher accuracy than at the beginning. So, the comparability between results of different laboratories becomes much better.
Tools for the characterization of secondary raw materials - status of leaching test application
(2018)
Leaching tests are used as tools for the assessment of environmental compatibility of granular waste materials with potential for reuse. The presentation gives an overview on available leaching tests, their field of application and relation to legislation. Furthermore robustness testing to elaborate optimized test conditions as well as performance characteristics from validation studies by BAM are addressed.
Tools for the characterization of secondary raw materials - status of leaching test application
(2018)
Leaching tests are used as tools for the assessment of environmental compatibility of granular waste materials with potential for reuse. The presentation gives an overview on available leaching tests, their field of application and relation to legislation. Furthermore robustness testing to elaborate optimized test conditions as well as performance characteristics from validation studies by BAM are addressed.
Potential use of incineration bottom ash in construction – Evaluation of environmental impact
(2018)
Around 5 million tonnes of MSWI bottom ash (BA) are generated per year in Germany. The incineration itself serves for metals as a concentrating and cleaning process. However, due to the almost exclusive wet extraction out of the furnace chamber the various metals are integrated in a heterogeneous and instable matrix. The metal recovery is therefore still a challenge relating the recovery rate and the purity, respectively. With state of the art treatment trains in Germany around 7.7% of ferrous metals and 1.3% of non-ferrous metals can be recovered out of MSWI BA. Large quantities of the mineral fraction are reused for sub-base material in road construction. A three month ageing period has been established as the common practice of further treatment of bottom ash before reuse applications. In the course of this aging the pH value of bottom ash decreases and contaminants are immobilized by processes like carbonation, hydration and oxidation. In particular leaching of heavy metals may be reduced to environmentally acceptable levels. However, the aging results in fixing of other valuable resources such as metals. Two laboratory scale lysimeters (30 cm in diameter) are being operated for more than three years aiming at the investigation of the long-term leaching behavior of a 0.25/45 mm mineral material obtained by treatment of MSWI bottom ash directly after incineration by a wet processing technology. Artificial rainwater is used as leachant (pH ≈ 6, ingredients NO32-, Cl-, SO42-, Na+, K+, Ca2+, Mg2+). The lysimeters are irrigated related to an average annual precipitation rate of 600 mm/a leading to a liquid to solid ratio (L/S) of about 0.7 per year of operation. Lysimeter tests are more adequate to simulate field scenarios and long-term leaching behavior in contrast to laboratory column tests particularly due to the unsaturated conditions and realistic contact time with the leachant.
In comparison to the leaching of bottom ash (aged for 3 month and non-treated) in column tests the chloride and sulfate leaching in the lysimeters was reduced as a consequence of the previous wet processing. The sulfate release kept almost constant limited by CaSO4 solubility up to an L/S of about 0.7 l/kg. The chloride concentrations dropped quickly, starting from 7300 mg/l, and are now at an almost constant level of 50 mg/l which is significantly above the chloride concentration in the leachant. After an initial decrease of the pH from about 10.4 to 8 at L/S of 0.2 l/kg, the pH increased again and leveled out around 9.8 up to L/S 2.2 l/kg. At an L/S of about 2.5 l/kg now, Cr, Cu are still released. The Cu and Cr concentrations in the leachates were about 760 µg/l and 90 µg/l respectively at the beginning. For both metals the concentration decreased to about 35 µg/l now. The Mo concentrations dropped steadily from 1300 to 250 µg/l, whereas V and Sb concentrations increased in the course of the tests. Fe and Mn are not detectable anymore. More results of this experimental study will be presented and discussed with a special focus on elements forming oxyanions.
Surface characterisation of biofilms by nearambient pressure X-ray photoelectron spectroscopy
(2018)
The XPS information depth of approximately 10 nm is in the same size range as the outer membrane of the gram-negative bacteria, which makes XPS a suitable tool for determining the elemental composition of the bacterial surface and monitor changes caused by outer stress like dehydration or exposure to antimicrobials. However, bacteria are inherently in a hydrated state, and therefore only compatible to ultra-high vacuum after extensive sample preparation, which may degrade the sample constituents. This issue is addressed by the development of near-ambient pressure (NAP)-XPS, which enables bacteria and biofilms to be characterised in their native wet state. Artificial biofilms, bacteria and biofilms of Escherichia coli have been characterised with the laboratory NAP-XPS instrument EnviroESCA from SPECS GmbH, at pressures ranging from high vacuum to 12 mbar, and in both humid and dry environment. By studying biological samples in their native wet state, new insight about composition and transport of drugs through cell membranes and the extracellular polymeric substance (EPS) of biofilms can be obtained.
The XPS information depth of approximately 10 nm is in the same size range as the outer membrane of the gram-negative bacteria, which makes XPS a suitable tool for determining the elemental composition of the bacterial surface and monitor changes caused by outer stress like dehydration or exposure to antimicrobials. However, bacteria are inherently in a hydrated state, and therefore only compatible to ultra-high vacuum after extensive sample preparation, which may degrade the sample constituents. This issue is addressed by the development of near-ambient pressure (NAP)-XPS, which enables bacteria and biofilms to be characterised in their native wet state. Artificial biofilms, bacteria and biofilms of Escherichia coli have been characterised with the laboratory NAP-XPS instrument EnviroESCA from SPECS GmbH, at pressures ranging from high vacuum to 12 mbar, and in both humid and dry environment. By studying biological samples in their native wet state, new insight about composition and transport of drugs through cell membranes and the extracellular polymeric substance (EPS) of biofilms can be obtained. In this contribution, the latest progress on biofilm characterisation by NAP-XPS will be presented, and measurement capabilities and limitations will be discussed.
A presentation held for the seminar "Ausgewählte analytische Methoden der Physik" hosted by Prof. Birgit Kanngießer at TU Berlin. The first part focus on depth-dependent XPS-measurements (XPS, synchrotron HAXPES) to obtain a concentration profile of iodine in an artificial biofilm. In the second part, NAP-XPS measurements of various bacterial samples are presented.
Near-ambient pressure XPS makes it possible to characterise samples not compatible to ultra-high vacuum, and enables the study of liquid-solid, gas-liquid and gas-solid interfaces. NAP-XPS meas-urements of biofilms, suspended nanoparticles and metal-organic frameworks were performed with EnviroESCA developed by SPECS.
An interesting application is surface characterisation of biofilms, which are bacterial communities embedded in a self-produced polysaccharide matrix. Various model systems ranging from pure polysaccharides of alginate to biofilms harvested directly from the growth medium have been char-acterised in humid conditions[1].
NAP-XPS also makes it possible to characterise nanoparticles in solution. Silver nanoparticles in aqueous solution were characterised and the Ag 3d-spectrum compared to spectra obtained of dried nanoparticles in UHV-conditions[2]. The binding energy of the Ag 3d-core level peak was shifted by 0,6 eV towards higher binding energy for suspended nanoparticles compared to the dried sample measured in UHV. This can be assigned to a change in surface potential at the water-nanoparticle interface.
Metal-organic frameworks (MOFs) are suitable materials for gas storage of small molecules due to their nanoporous, crystalline structure. However, instability in humidity remains an issue for many types of MOFs. XPS-measurements of the MOF-structure HKUST-1 were performed in various NAP-conditions to assess the stability of the sample and its interaction with the gas molecules as water, methanol and pyridine.
Synchrotron XPS in the soft-X-ray regime is suitable for the detection of light elements commonly found in biological samples. Various model systems of biofilms have been developed and characterised at synchrotron- and lab-based facilities. By obtaining the chemical composition at various information depths, the vertical distribution of iodine in an artificial biofilm have been determined.
X-ray photoelectron spectroscopy (XPS) provides elemental and chemical information from the outermost ~10 nm of the sample surface. This is in the same order of magnitude as the thickness of the outer bacterial membrane of gram-negative bacteria, as well as outer membrane molecules as exopolysaccharides and lipopolysaccharides, commonly attached to the cell surface. With the development of near-ambient pressure (NAP)-XPS, bacteria can be analysed with minimal sample preparation.
EnviroESCA is a laboratory based NAP-XPS instrument, equipped with a monochromated Al Kα radiation source and a differentially pumped energy analyser connected to an exchangeable sample environment. It allows for measurements in various gas-atmospheres, including water vapor, which makes it possible to characterise bacteria and other biological samples close to their natural, hydrated state. Artificial model-biofilms of exopolysaccharides, planktonic Pseudomonas Fluorescens and biofilms of Escherichia Coli have been characterised in hydrated and dried state.
High-resolution XPS-spectra from carbon, oxygen, nitrogen and phosphorous can be assigned to carbohydrates, lipids and proteins in general agreement with literature. Especially the carbon 1s peak is of interest. A series of measurements of an E. coli biofilm from 11 mbar in humid environment to 1 mbar air reveal changes in the C1s peak, which suggests that the bacterial surface undergo substantial Change.
This application note presents how EnviroESCA can be used to analyze E. coli biofilms on silicon under near ambient pressure conditions in various states of hydration. Such investigations of the outer bacterial cell surface in their hydrated state are essential for studying biological interfaces at work.
Microbiologically influence corrosion (MIC) has become a big concern due the increased usage of different metals by our society. Microorganisms can use metal as an electron donor, causing unpredictable but serious damages. Nowadays it is known that besides sulfate reducing bacteria (SRB), other microorganisms including acetogens, iron oxidizers and methanogens can also induce MIC. Current studies related to methanogen-induced MIC (MI-MIC) mainly focused on environmental isolates from the oil and gas industry (e.g. Methanococcus maripaludis) with industrial materials e.g. iron. However, MI-MIC can occur in many other environments as well, including the oral cavity. Methanobrevibacter oralis is a methanogen isolated from the human oral cavity and was found more frequently in patients suffering from peri-implantitis/periodontitis. Titanium-implants removed from those patients have also showed clear signs of corrosion. The aim of our study is to establish and analyze corrosion potentials of dental metals (e.g. titanium) by oral methanogens. Periodontal pockets samples from patients suffering from periodontitis/peri-implantitis were taken for methanogenic and SRB enrichments. Stainless steel, pure titanium or Ti-6Al-4V alloy was used for corrosion studies. Corrosion rates and methane production were measured using weight-loss method and gas chromatography, respectively. Metal surfaces were visualized with scanning electron microscopy. Microbial communities in the dental pockets of healthy people and patients will be compared using 16S rRNA amplicon sequencing. Overall, this is the first study investigating the susceptibility of different dental implant materials to corrosion using human-related Archaea. The outcomes of this study can be further explored for a variety of clinical applications.
Microbiologically influenced corrosion (MIC) is the deterioration of metals due to the metabolic activities of microorganisms. Microorganisms can take electrons directly from the metal surface (EMIC) thereby causing corrosion. Well known culprits of EMIC are: sulfate-reducing bacteria (SRB), acetogens and methanogens.
Microbially influenced corrosion (MIC) of iron is usually attributed to sulfate-reducing microorganisms (SRM) which act upon the metal by the reactiveness of hydrogen sulfide, and by withdrawal of the available electrons (Fe → Fe²⁺ + 2e⁻ ; E° = 0.47 V) in electrical contact through surface attachment. Also methanogenic archaea are supposed to cause MIC. Because they do not produce hydrogen sulfide, withdrawal of electrons may be their main corrosive mechanism; however, mechanistic details and kinetics of the overall process are poorly understood. Precipitation of siderite (4Fe + 5HCO₃⁻ + 5H⁺ → 4FeCO₃ + CH₄ + 3H₂O) can lead to an insulating layer on the metal surface and lower the corrosion rate. Still, the extent of FeCO₃ precipitation may be significantly influenced by environmental conditions such as pH and advective processes.
To investigate the corrosive potential of methanogens, we studied strains isolated from marine sediments (Methanococcus maripaludis 14266, 2067, Methanobacterium-affiliated strain IM1), crude oil tanks (Methanococcus maripaludis Mic1c10, KA1) and the oral cavity (Methanobrevibacter oralis) in a closed (batch) culture, and in a sand-packed flow-through cell with pH control and simulation of a fluctuating environment. Results indicate that the rates of iron corrosion due to coupled methanogenesis (up to 0.3 mm/yr) are comparable to that caused by SRM. Surface analyses of the metal showed severe pitting. Such knowledge and deeper understanding also from an electrokinetic point of view may not only provide further models in microbial electrophysiology, but also contribute to mitigation strategies in MIC.
Microbially influenced corrosion (MIC) of iron is usually attributed to sulfate-reducing microorganisms (SRM) which act upon the metal by the reactiveness of hydrogen sulfide, and by withdrawal of the available electrons in electrical contact with the metal. Methanogenic archaea can also cause MIC by directly withdrawing electrons from the iron surface for methanogenesis. However, the mechanistic details and kinetics of the overall process are poorly understood. Precipitation of siderite, a by-product of methanogenesis, (4Fe + 5HCO3 + 5H+ 4FeCO3 + CH4 + 3H2O) can lead to an insulating layer on the metal surface and lower the corrosion rate. Still, the extent of FeCO3 precipitation may be significantly influenced by environmental conditions such as pH and advective processes.
Microbially influenced corrosion (MIC) of iron is usually attributed to sulfate-reducing microorganisms (SRM) which act upon the metal by the reactiveness of hydrogen sulfide, and by withdrawal of the available electrons in electrical contact with the metal. Methanogenic archaea are supposed to cause MIC. Because they do not produce hydrogen sulfide, withdrawal of electrons may be their main corrosive mechanism; however, mechanistic details and kinetics of the overall process are poorly understood. Precipitation of siderite (4Fe + 5HCO3 + 5H+ 4FeCO3 + CH4 + 3H2O) can lead to an insulating layer on the metal surface and lower the corrosion rate. Still, the extent of FeCO3 precipitation may be significantly influenced by environmental conditions such as pH and advective processes.
Microbially influenced corrosion (MIC) of iron is usually attributed to sulfate-reducing microorganisms (SRM) which act upon the metal by the reactiveness of hydrogen sulfide, and by withdrawal of the available electrons in electrical contact with the metal. Methanogenic archaea are supposed to cause MIC. Because they do not produce hydrogen sulfide, withdrawal of electrons may be their main corrosive mechanism; however, mechanistic details and kinetics of the overall process are poorly understood. Precipitation of siderite (4Fe + 5HCO3 + 5H+ 4FeCO3 + CH4 + 3H2O) can lead to an insulating layer on the metal surface and lower the corrosion rate. Still, the extent of FeCO3 precipitation may be significantly influenced by environmental conditions such as pH and advective processes.
Microbiologically influenced corrosion (MIC) of iron is usually attributed to sulfate-reducing microorganisms, either chemically (formation of hydrogen sulfide) or electrically (direct electron uptake). Methanogenetic Archaea are also known to be involved in iron corrosion, forming a multi-species biofilm on corroding metallic structures. However, mechanistic details and kinetics of the overall process in methanogen-induced MIC are poorly understood.
The particle size distribution is considered the most relevant information for nanoscale property identification and material characterization. The current OECD test guideline on particle size and size distribution (TG 110) is not applicable to ‘nano-sized’ objects. In this project we thus develop a new OECD test guideline for the measurement of the size and size distribution of particles and fibers with at least one dimension in the range of 1 - 1000 nm. A fiber is defined as an object having an aspect ratio of length/diameter l/d >3. The width and length of each fiber should be measured concurrently.
In order to measure the particle size distributions, many techniques are available. 9 methods for particles and 2 methods for fibres have been tested in a prevalidation study and appropriate methods will be compared in an interlaboratory round robin test starting in February 2019.
The particle size distribution is considered the most relevant information for nanoscale property identification and material characterization. The current OECD test guideline on particle size and size distribution (TG 110) is not applicable to ‘nano-sized’ objects. In this project we thus develop a new OECD test guideline for the measurement of the size and size distribution of particles and fibers with at least one dimension in the range of 1 - 1000 nm. A fiber is defined as an object having an aspect ratio of length/diameter l/d >3. The width and length of each fiber should be measured concurrently.
In order to measure the particle size distributions, many techniques are available. 9 methods for particles and 2 methods for fibres have been tested in a prevalidation study and appropriate methods will be compared in an interlaboratory round robin test starting in February 2019.
The new OECD test guideline will address the following four main steps in the determination of the length and width distributions of fibers: sample preparation, image acquisition, data evaluation and uncertainty analysis. As the sample preparation has to be optimized for each material, general quality criteria will be given in the protocol. For full visibility of a fiber the appropriate resolution has to be chosen. In the data evaluation the length and diameter of each fiber will be determined concurrently to allow for application of different regulatory definitions. The quality of the results critically depends on the sample preparation as well as the data evaluation. In this step the classification rules have to be formulated and followed accurately in order to optimize reproducibility of the method. The SOP will be validated in an international round robin test, which is planned for 2018/2019.
Without precaution, insects may cause serious damage to Museum collections. Quarantine of potentially infested objects can be logistically challenging. Anoxia under controlled nitrogen atmosphere is a most compatible but also time-consuming method to eradicate insect pests in all kinds of different materials. Treatment results are usually effected by duration, temperature, humidity and residual oxygen content. During a two-year research project, 34 relevant pest insect species of all developmental stages were tested in several different materials (wood, paper, wool) to monitor treatment success and to determine optimum treatment parameters. Duration of Treatment ranged from one to three weeks at temperatures of 20 - 27 °C.
As expected, results showed significant differences in mortality among tested species. Highest tolerance of hypoxic conditions was found in older larvae of Hylotrupes bajulus. However, this species is an unlikely museum pest. Anobiids and other wood boring beetles are more often an issue related to cultural heritage. Tested imbedding materials in general had no mortality lowering influence. A combination of three weeks exposure time at up to 0.5 % residual oxygen and at 24 °C and 50 % RH is recommended for infested artefacts.
Zearalenone (ZEN) and its sulfate and glucoside conjugates have been detected in (a broad variety of) food and feed commodities1. Both conjugated derivatives are formed as part of fungal or plant secondary metabolism and thus, belong to the group of modified mycotoxins2. After consumption of contaminated foodstuff, the conjugates can be hydrolyzed by human intestinal microbiota leading to liberation of ZEN that implies an underestimation of the true ZEN exposure. In order to include ZEN conjugates in routine analysis, as well as for toxicological investigation reliable standards are needed.
The objective of the present study was to develop a simple and economic method for biosynthesis of ZEN conjugates. Preceding experiments on the biotransformation of ZEN by Rhizopus and Aspergillus species showed a mixed metabolite formation3. Therefore, these known ZEN conjugating fungal strains were screened for their potential to selectively synthesize the ZEN derivatives ZEN-14-sulfate (Z14S), ZEN-14-glucoside (Z14G) and ZEN-16-glucoside (Z16G). The screening was conducted by adding ZEN to liquid fungal cultures. Cultivation conditions and ZEN incubation time were varied. All media samples were analyzed for metabolite formation by HPLC-MS/MS. Z14S was exclusively formed by A. oryzae. Under optimized conditions a specific biosynthesis of Z14G by R. oryzae and Z16G by R. oligosporus was achieved. After liquid-liquid-extraction and preparative chromatographic cleanup 1H-NMR purities of ≥ 73% for Z14S, ≥ 82% for Z14G and ≥ 50% for Z16G were obtained. In addition, a consecutive biosynthesis was developed by first using Fusarium graminearum for ZEN biosynthesis on rice based liquid medium. After inactivation of Fusarium the subsequent conjugation reaction was conducted utilizing Aspergillus and Rhizopus species under the various optimized conditions.
In this study an easy and cost-efficient biosynthesis for Z14S, Z14G and Z16G was developed. The developed biosynthesis could be also used for other metabolites like ZEL conjugates. Our results of the in vitro screening indicate also the formation of a ZEL-glucoside and α ZEL-sulfate as major metabolites by R. oryzae. In sum, under optimized cultivation conditions fungi can be easily utilized for a targeted and stereospecific synthesis of ZEN conjugates.
Bee populations have declined significantly in recent years and this is thought to be attributable at least in part to the (re-)emergence of viruses. These viruses are predominantly positive single stranded (+ss) RNA viruses belonging to the Picornavirales. Managed honeybees are often infested with the invasive mite, Varroa destructor, which vectors RNA viruses including Deformed wing virus (DWV, family Iflaviridae): a leading culprit of colony losses. Many bee viruses have been sequenced and structural features are now available for viruses such as DWV. DWV consists of at least 3 distinct genotypes, two of which have been shown to be differentially virulent in honeybees. Molecular studies have demonstrated that DWV has a mean evolutionary rate of 1.35 x 10-3 per site per year. For such viruses – in contrast to their eukaryotic hosts – ecological and evolutionary timescales significantly overlap. This rapid evolution allows RNA viruses to adapt quickly to novel host environments with recombination representing an additional key source of genetic variation. Interestingly, recombination between genotypes of DWV has recently been shown to be a common occurrence in honeybees. A challenge has been to develop bioinformatics tools that can accurately reconstruct viral haplotypes – including recombinants – from heterogenous high-throughput sequence data.
The impact of blood-to-blood Varroa destructor transmission on virus evolution represents an important question in bee virus research. Due to the nature of the V. destructor life cycle, predictions can be made about the potential impact of the mite on virus virulence evolution. Specifically, the developing honeybee host pupa should remain alive until close to the completion of metamorphosis to provide sufficient time for successful mite reproduction, including offspring mating. For optimal transmission, any virus found in a mature and mated daughter mite will hold a significant selective advantage over a virus found in an immature or unmated daughter mite – placing a cost on virus virulence that impacts honeybee pupae before mites can mate. On the other side, viruses replicating too slowly and with delayed virulence effects will hold a selective handicap because fewer transmission units will be found in mated mites. I have hypothesized that the evolution of virus virulence shifted following the arrival of V. destructor, with viruses, including recombinants and/or specific viral genotypes being selected for a level of virulence in pupae (and likely also in adults) that maximises R0, which represents the basic reproductive number of the virus in the host population. R0 is defined by the number of subsequent infections caused by a single infection and it must be greater than 1 for an infection to spread in a population. It is enhanced by maximising the number of transmission units passed to uninfected susceptible vectors, and ultimately hosts (Figure 1).
Honeybee viruses are also shared with sympatric wild bees and viral prevalence and sequence data indicate frequent virus transmission between managed and wild bee species. In addition to infecting the western honeybee (Apis mellifera), DWV can infect other Asian honeybee species such as Apis ceranae. Outside of honeybees, DWV has been found widely in bumblebees, including solitary bees and wasps and there is evidence that it can actively replicate in several Bombus and solitary bee species. Whether the arrival of the V. destructor mite in A. mellifera has driven viral emergence in non-Apis bees is a target of ongoing research.
Technical advances in the study of molecular evolution have crystallized the fundamental insight that many bee pathogens evolve and adapt over timescales that overlap with host ecology. At the same time, the role played by bee host community ecology is increasingly being appreciated in host-parasite interactions. Here, we focus on Deformed wing virus (DWV) and present recent studies exploring the link between virulence, DWV genetic diversity and changes to host ecology - namely the arrival of an invasive ectoparasite, the Varroa destructor mite, which vectors viruses between honeybees. Specifically, we show how V. destructor may have created conditions for the emergence of more virulent strains of DWV in the western honeybee, Apis mellifera. We present a molecule-to-ecology framework to help interpret findings and to guide future hypotheses, emphasizing the role of molecular interactions between viruses and host immunity as drivers of change at the bee population level.
Our understanding of RNA viruses from edible insects is minimal at best, with studies largely focusing on model insect species and those associated with obvious signs of disease. This represents a considerable gap in understanding, given the growing role of insects as a source of food and feed, as well as the more general relevance of insects in agriculture and health. Illness due to entomophagy is rare but well documented, including fatal cases following the consumption of termites. Termites are eaten commonly in tropical Asia, Africa and South America, and are among the insects with the highest recorded fat content. There are many species of termites, with a wide range of diets and habitats centering around the consumption of wood and soil substrates. In this study, we report the results from a survey of more than 30 cockroach and termite transcriptomes, with the aim of understanding the diversity and evolution of RNA viruses as well as other potentially pathogenic organisms that are associated with this relevant but somewhat overlooked group of insects. We discuss our results in the context of the possible zoonotic risk posed by insects, as well as in the context of emerging viral and other disease threats that may face insects being reared at industrial scales.
The termites are a derived eusocial lineage of otherwise non-social cockroaches. Understanding the proximal and ultimate drivers of this major evolutionary transition represents an important goal in biology. One outstanding question concerns the evolution of termite immunity, which is thought to have undergone broad-sweeping adaptations in order to enhance group-level immune protection. To understand the evolutionary origins of termite immunity, we conducted qualitative and quantitative transcriptome analyses along a gradient of sociality. Firstly, we aimed to identify large-scale genetic shifts in immune traits linked to eusociality by comparing immune gene repertoires in solitary and subsocial cockroaches and across a range of eusocial termite lineages. Secondly, we compared the responses of a solitary cockroach, a subsocial wood-roach and different castes of a lower termite species to a non-specific immune challenge, in order to understand how sociality may have influenced the evolution of immune gene regulation. Firstly, we found that termites have a broadly representative repertoire of canonical immune genes as compared to gregarious cockroaches and subsocial wood-roaches. Secondly, with respect to immune challenge, the solitary cockroach and the subsocial wood-roach displayed a similarly comprehensive induced response, while the termite response was considerably dampened by comparison and strongly influenced by caste; with reproductives displaying a generally higher constitutive level of immune-gene expression compared to sterile castes. In summary we did not find termite eusociality to be associated with significant changes in immune gene diversity, but rather to be linked with significant modifications to the regulation of immunity following the origin of division of labour.
X-ray fluorescence analysis, due to its non-destructive nature and ist suitability to work with historic objects in situ, quickly became one of the most important methods for the evaluation of iron-gall ink. The main advantage of this qualitative and semi-quantitative method is that it makes it easy to differentiate between inks, based on the assumption that the differences result from the manufacture of the ink.
This work explores the question whether the ink ‘fingerprint’ results strictly from the elemental composition of the basic ink ingredients even if it is stored in vessels made of metals or metal alloys. In addition, we tested and compared the performance of three different XRF spectrometers.
We prepared various lab-grade inks according to historical ink recipes and measured the metal content of the ink deposited on sized cotton Linters paper with three types of XRF spectrometers: a simple hand-held device with an interaction spot of 4 mm and two devices equipped with poly-capillary Xray optics for line scanning and imaging. Since the exact elemental mass composition of the non-aged ink samples was known, we were able to evaluate the accuracy of the research procedure. Lab-grade inks were then aged in the metal jars imitating inkwells. The aging of the inks in the metal containers resulted in the significant change of the primary inks fingerprint as opposed to that of the control inks stored in glass containers. This effect was independently confirmed by the measurements conducted with every instrument we used. We will present a brief comparison of the results 43 achieved when using different spectrometers and a possible hypothesis explaining the processes that occurred.
The mayor part of the Berlin collection `Manuscripta Americana´ consists of documents compiled by Alexander von Humboldt. The diversity of the written fragments and their shape presume few accordance concerning purpose, place or time of production. Even before the papers came to Berlin, manuscripts were copied, divided and collected by famous scholars also in Mexico. Back in Europe, Humboldt visited further collections and published the detected hieroglyphic writings altogether with cultural objects and landscapes in the book `Vues des Cordillères et Monuments des Peuples Indigènes de l`Amerique, voyage de Humboldt et Bonpland´, labelled corresponding to the cultural background.
By scientific and culture historical analyses, the Humboldt codices are revised and related to each other and others. In especial, the compilation of fragments shown on plate 36 in the book is looked at more closely. Whereas most of the plates represent only one sheet of painted amate paper, here segments with details of seven fragments are put together. It is tried to comprehend why these pieces are on one plate.
To find out a common feature of the documents, their content and represented form are compared. A correlation between some pieces is obvious and thus is also expected in the manufacture of the codices, in the kind of material used. For this the inks and colors are determined by scientific analysis including XRF-, Raman-, FTIR- and VIS- spectroscopy. Special material is detected and related to results of former analysis of codices written during the colonial period of America. Herewith a clear cultural assignment of the manuscripts is performed.
A relation of the fragments represented on plate 36 among each other and to other codices of the collection Manuscripta Americana in Berlin is discussed. Also, if the manuscripts are unique `originals´ or represent fragments of documents, whose corresponding pieces maybe found- in the best case- incorporated in other collections. We detected, that at least two of them can be put into relation to similar manuscripts deposited in Mexico. Further material analysis should clarify, if the corresponding pieces are identical- from one document, or if one of them is a copy. Herewith an approximation to authenticity features and history is issued.
Antimicrobial resistance is a major threat to human health. The prevalence of multi-drug resistant (MDR) bacteria is predicted to increase in the future requiring robust control strategies. One cornerstone to prevent the spread of MDR bacteria in clinical settings is the application of disinfectants to improve hygiene standards. However, bacteria can evolve resistance to disinfectants, which in turn can confer cross-resistance to antibiotics. Additionally, clonal bacterial populations can display phenotypic heterogeneity with respect to the tolerance of antibiotic stress leading to a prolonged survival of a sub-population; this phenomenon is termed persistence. Persistence to antibiotics is an evolvable trait and can serve as a stepping stone for the evolution of genetically encoded resistance. Until now, there is a lack of systematic studies that investigate if bacterial populations establish persister subpopulations that tolerate disinfectant stress longer than the majority of the population. Our results in E. coli indicate that persistence is a bacterial survival strategy against benzalkonium chloride, a widely used disinfectant.
In future experiments, we will investigate the evolution of persistence in the face of fluctuating exposure to disinfectants and whether persistence facilitates resistance against disinfectants. Lastly, we will test how tolerance and resistance against disinfectants affects susceptibility against antibiotics.
Infections with multi-resistant Gram negative pathogens are a major threat to our health system. In order to serve the needs in antibiotics development we selected untapped bioresources and implemented high throughput approaches suitable for the discovery of strains producing antibiotics with anti-Gram negative activity.
Our approaches rely on the hypothesis that Termite associated bacteria are likely to produce potent antibiotics to defend their hosts against entomopathogenic microorganisms. Termite nests and guts harbor suitable, highly diverse microbiomes in which bacterial taxa are present known to potentially produce natural compounds. In
a first step the diversity of Coptotermes species nest microbiomes was assessed carefully by using 16S rDNA amplicon sequencing on the Illumina MiSeq platform and nest material was selected to retrieve viable cells by using Nycodenz density gradient centrifugation. In order to analyze the diversity of the culturable termite nest microbiome, bacterial cells were either distributed in 384-well plates
(approach 1) or encapsulated in small spheric agarose beads by an high throughput microfluidics technique (approach 2). Cultures obtained from approach 1 were scaled-up in 96-well Duetz-systems for characterization of diversity and for rapid supernatant screening using the bioluminescence-labeled E. coli pFU166. The generated
droplets of approach 2 simultaneously received a small population of GFP-tagged Gram negative screening cells and were sorted for low fluorescence using FACS. After elimination of redundancy we performed a fast scale-up of active strains.
Implementation of this pipeline allows us to prioritize antibiotics producing strains in a ultra-high throughput fashion and by cultivation of broad diversity in our approches.
Environmental sustainability and –stability of Materials concerning the Migration of pollutants
(2018)
MaUS is an acronym for ”Material und Umweltsimulationen“. Plastics are in the focus of environmental politics due to their long-term behaviour and therefore to their persistence. Not only that they appear as visible contaminants in the sea and on the beach, but their unknown behaviour concerning their additives as well as the related transformation products are anxious. Therefore, we wish to establish a certified reference method to provide a method for testing plastics.
Aim of this project is the development of fast motion standard reference methods for testing plastics regarding to their environmental compatibility. To establish these testing methods, we use polystyrene (PS) and polypropylene (PP) with environmental relevant brominated flame retardants, known for their persistent bioaccumulative and toxic (PBT) properties. In case of PS the material contains 1 wt% of 1,2,5,6,9,10-hexabromocyclododecan (HBCD) and in case of PP 0.1 wt% bromodiphenylether (BDE-209), which is known as a substance of very high concern (SVHC). Furthermore, we use polycarbonate (PC), which is still used as material in baby flasks and releases Bisphenol A (BPA), an estrogenic active substance.
As an additional material PTFE is used for its importance as a source for two ubiquitous environmental substances (PFOS and PFOA), whose toxicological effects are still incompletely known.
The focus in this current work is set on the transfer of potential pollutants out of applied materials mentioned above into environmental compartments like water or soil. Here an accelerated aging concept should be developed to shortened time consuming natural processes. For these resulting simulations we use a programmable weathering chamber with dry and wet periods and with high and low temperatures. These programmes run for several weeks and according to a defined sampling schedule we take water samples, run a clean-up procedure by SPE (Molecular imprinted polymers (MiPs) resp. polymer-based cartridges (Waters Oasis HLB)) and analyse them by HPLC-UV resp. LC-MS/MS. Of most interest in case of flame retardants are photocatalytic transformation products. Therefore, we conduct a non-target-screening resp. a suspected target-screening by LC-MS/MS and HRMS.