Life expectancy, family constellation and stress in giant mole-rats (Fukomys mechowii).

Giant mole-rats (Fukomys mechowii) are remarkably long-lived subterranean rodents (maximum recorded lifespan as reported here greater than 26 years) that live in families with one reproductive pair (breeders) and their non-reproductive offspring (non-breeders). Previous studies have shown that breeders live on average approximately twice as long as non-breeders, a finding contradicting the classic trade-off between reproduction and lifespan. Because recent evidence points to the hypothalamic-pituitary-adrenal axis as playing an important role in shaping the pace of ageing in mole-rats, we analysed the influence of the social environment of giant mole-rats on intrafamilial aggression levels, indicators of long-term stress, and, ultimately, mortality. Behavioural data indicated that family constellation, especially the presence or the absence of parents, influences agonistic behaviour. As a measure of long-term stress, we established a non-invasive method of extracting and measuring cortisol from hair of giant mole-rats. Interestingly, orphaned non-breeders exhibited significantly lower levels of cortisol and lower mortality rates than did non-breeders living with both parents. Because hypercortisolism is harmful in the long-term, intrafamilial stress could help explain the earlier onset of senescence in non-breeders, resulting in a shorter lifespan. Our findings suggest that the social environment should be considered as a further factor in ageing studies involving group-living animals. This article is part of the theme issue 'Ageing and sociality: why, when and how does sociality change ageing patterns?'

A centrifuge tube reactor for the determination of bacterial methane oxidation enrichment factors without influence of diffusion related isotope fractionation.

Biotransformation of methane at landfill sites can be estimated by applying compound specific stable isotope analysis of methane from the anaerobic and the cover layer surface zone. Next to these two input parameters, merely the knowledge of the carbon isotopic fractionation of the bacterial methane oxidation in terms of the enrichment factor (ε) is required. However, many factors and conditions have been described to affect ε. These include temperature, the applied landfill cover, the type of expressed methane monooxygenase (MMO), and cell density. In this work we investigated the microbial methane oxidation with respect to temperature and type of methanotrophic enrichment culture. A newly designed setup was used to overcome potential CH4-substrate limitations such as diffusion that could affect the determined values of ε by improper and inhomogeneous mixing. The isotopic fractionation was determined based on the stable carbon isotope analysis of methane and carbon dioxide. The obtained value for isotopic fractionation was ε22°C = -0.0136 ±â€¯0.0036. Also for the first time, bulk stable isotope analysis of bacterial cell mass was performed by flow injection analysis isotope ratio mass spectrometry.

The monogenean Paradiplozoon ichthyoxanthon behaves like a micropredator on two of its hosts, as indicated by stable isotopes.

The analysis of stable isotopes of carbon and nitrogen has been used as a fingerprint for understanding the trophic interactions of organisms. Most of these studies have been applied to free-living organisms, while parasites have largely been neglected. Studies dealing with parasites so far have assessed the carbon and nitrogen signatures in endoparasites or ectoparasites of different hosts, without showing general trends concerning the nutritional relationships within host-parasite associations. Moreover, in most cases such systems involved a single host and parasite species. The present study is therefore the first to detail the trophic interactions of a freshwater monogenean-host model using δ13C and δ15N, where a single monogenean species infects two distinctly different hosts. Host fishes, Labeobarbus aeneus and Labeobarbus kimberleyensis from the Vaal Dam, South Africa, were assessed for the monogenean parasite Paradiplozoon ichthyoxanthon, individuals of which were removed from the gills of the hosts. The parasites and host muscle samples were analysed for signatures of δ13C and δ15N using an elemental analyser connected to an isotope ratio mass spectrometer. Host fish appear to use partly different food sources, with L. aeneus having slightly elevated δ13C signatures compared to L. kimberleyensis, and showed only small differences with regard to their nitrogen signatures, suggesting that both species range on the same trophic level. Carbon and nitrogen signatures in P. ichthyoxanthon showed that the parasites mirrored the small differences in dietary carbon sources of the host but, according to δ15N signatures, the parasite ranged on a higher trophic level than the hosts. This relationship resembles predator-prey relationships and therefore suggests that P. ichthyoxanthon might act as a micropredator, similar to blood-sucking arthropods such as mites and fleas.

A novel tool for stable nitrogen isotope analysis in aqueous samples.

RATIONALE: Bulk stable isotope analysis (BSIA) of dissolved matter (e.g. dissolved organic carbon, total nitrogen bound (TNb ), etc.) is of particular importance since this pool is a prime conduit in the cycling of N and C. Studying the two elemental pools is of importance, as transformation and transport processes of N and C are inextricably linked in all biologically mediated systems. No system able to analyze natural abundance stable carbon and nitrogen isotope composition in aqueous samples (without offline sample preparation) and simultaneously has been reported so far. Extension of the high-temperature combustion (HTC) system, to be capable of measuring TNb stable nitrogen isotope composition, is described in this study. METHODS: To extend the TOC analyzer to be capable of measuring TNb , modifications from the HTC high-performance liquid chromatography/isotope ratio mass spectrometry (HPLC/IRMS) interface were implemented and expanded. A reduction reactor for conversion of NOx into N2 was implemented into the new developed system. The extension addresses mainly the development of the focusing unit for nitrogen and a degassing device for online separation of TNb from molecular nitrogen (N2 ) prior to injection. RESULTS: The proof of principle of the system was demonstrated with different compound solutions. In this initial testing, the δ15 NAIR-N2 values of the tested compounds were determined with precision and trueness of typically ≤0.5‰. Good results (u ≤ 0.5‰) could be achieved down to a TNb concentration of 40 mgN/L and acceptable results (u ≤ 1.0‰) down to 5 mgN/L. In addition, the development resulted in the first system reported to be suitable for simultaneous and direct δ13 C and δ15 N BSIA of aqueous samples. CONCLUSIONS: The development resulted in the first system shown to be suitable for both δ13 C and δ15 N direct BSIA in aqueous samples. This system could open up new possibilities in SIA-based research fields. Copyright © 2016 John Wiley & Sons, Ltd.

Fate and behaviour of diclofenac during hydrothermal carbonization.

Hydrothermal carbonization (HTC) has become an esteemed method to convert sewage sludge into biochar. Besides dewatering and disinfection the process is suggested to reduce the micropollutant load, which would be beneficial for the use of biochar as fertilizer. This study was designed to examine reduction of micropollutants and formation of transformation products during HTC using the example of diclofenac. We investigated compounds' removal at HTC conditions in inert experiments and in real samples. Results showed that HTC temperature (>190 °C) and pressure (∼15 bar) have the potential to fully degrade diclofenac in inert experiments and spiked sewage sludge (>99%) within 1 h. However, interfering effects hinder full removal in native samples resulting in 44% remaining diclofenac. Additionally, a combination of suspected-target and non-target analysis using LC-MS/MS and LC-HRMS resulted in the determination of six transformation products. These products have been reported in biochar from HTC for the first time, although other studies described them for other processes like advanced oxidation. Based on the detected transformation products, we proposed a degradation mechanism reflecting HTC reactions such as dehydroxylation and decarboxylation.

A novel high-temperature combustion interface for compound-specific stable isotope analysis of carbon and nitrogen via high-performance liquid chromatography/isotope ratio mass spectrometry.

RATIONALE: In aqueous samples compound-specific stable isotope analysis (CSIA) plays an important role. No direct method (without sample preparation) for stable nitrogen isotope analysis (δ(15) N SIA) of non-volatile compounds is known yet. The development of a novel HPLC/IRMS interface based on high-temperature combustion (HTC) for both δ(13) C and δ(15) N CSIA and its proof of principle are described in this study. METHODS: To hyphenate high-performance liquid chromatography (HPLC) with isotope ratio mass spectrometry (IRMS) a modified high-temperature combustion total organic carbon analyzer (HTC TOC) was used. A system to handle a continuously large amount of water (three-step drying system), favorable carrier and reaction gas mix and flow, an efficient high-temperature-based oxidation and subsequent reduction system and a collimated beam transfer system were the main requirements to achieve the necessary performance. RESULTS: The proof of principle with caffeine solutions of the system succeeded. In this initial testing, both δ(13) C and δ(15) N values of tested compounds were determined with precision and trueness of ≤0.5 ‰. Further tests resulted in lower working limit values of 3.5 µgC for δ(13) C SIA and 20 µgN for δ(15) N SIA, considering an accuracy of ±0.5 ‰ as acceptable. CONCLUSIONS: The development of a novel HPLC/IRMS interface resulted in the first system reported to be suitable for both δ(13) C and δ(15) N direct CSIA of non-volatile compounds. This highly efficient system will probably open up new possibilities in SIA-based research fields. Copyright © 2016 John Wiley & Sons, Ltd.

Pharmaceutical load in sewage sludge and biochar produced by hydrothermal carbonization.

We investigated the removal of twelve pharmaceuticals in sewage sludge by hydrothermal carbonization (HTC), which has emerged as a technology for improving the quality of organic waste materials producing a valuable biochar material. In this study, the HTC converted sewage sludge samples to a biochar product within 4h at a temperature of 210 °C and a resulting pressure of about 15 bar. Initial pharmaceutical load of the sewage sludge was investigated as well as the residual concentrations in biochar produced from spiked and eight native sewage sludge samples from three waste water treatment plants. Additionally, the solid contents of source material and product were compared, which showed a considerable increase of the solid content after filtration by HTC. All pharmaceuticals except sulfamethoxazole, which remained below the limit of quantification, frequently occurred in the investigated sewage sludges in the µg/kg dry matter (DM) range. Diclofenac, carbamazepine, metoprolol and propranolol were detected in all sludge samples with a maximum concentration of 800 µg/kgDM for metoprolol. HTC was investigated regarding its contaminant removal efficiency using spiked sewage sludge. Pharmaceutical concentrations were reduced for seven compounds by 39% (metoprolol) to≥97% (carbamazepine). In native biochar samples the four compounds phenazone, carbamazepine, metoprolol and propranolol were detected, which confirmed that the HTC process can reduce the load of micropollutants. In contrast to the other investigated compounds phenazone concentration increased, which was further addressed in thermal behaviour studies including three structurally similar potential precursors.

Determination of pharmaceuticals in sewage sludge and biochar from hydrothermal carbonization using different quantification approaches and matrix effect studies.

Producing valuable biochar from waste materials using thermal processes like hydrothermal carbonization (HTC) has gained attention in recent years. However, the fate of micropollutants present in these waste sources have been neglected, although they might entail the risk of environmental pollution. Thus, an HPLC-MS/MS method was developed for 12 pharmaceuticals to determine the micropollutant load of biochar, which was made from sewage sludge via HTC within 4 h at 210 °C. Pressurized liquid extraction was applied to extract the compounds. Because of the high load of co-extracted matter, matrix effects in HPLC-MS/MS were investigated using matrix effect profiles. Interfering compounds suppressed 50% of the phenazone signal in sewage sludge and 70% in biochar, for example. The quantification approaches external calibration, internal standard analysis, and standard addition were compared considering recovery rates, standard deviations, and measurement uncertainties. The external analysis resulted in decreased or enhanced recovery rates. Spiking before LC-MS/MS compensated instrumental matrix effects. Still, recovery rates remained below 70% for most compounds because this approach neglects sample losses during the extraction. Internal standards compensated for the matrix effects sufficiently for up to five compounds. The standard addition over the whole procedure proved to compensate for the matrix effects for 11 compounds and achieved recovery rates between 85 and 125%. Additionally, results showed good reproducibility and validity. Only sulfamethoxazole recovery rate remained below 70% in sewage sludge. Real sample analysis showed that three pharmaceuticals were detected in the biochar, while the corresponding sewage sludge source contained 8 of the investigated compounds.

A novel high-temperature combustion based system for stable isotope analysis of dissolved organic carbon in aqueous samples. I: development and validation.

RATIONALE: Traditionally, dissolved organic carbon (DOC) stable isotope analysis (SIA) is performed using either offline sample preparation followed by elemental analyzer/isotope ratio mass spectrometry (EA/IRMS) or a wet chemical oxidation (WCO)-based device coupled to an isotope ratio mass spectrometer. The first method is time-consuming and laborious. The second involves the risks of underestimation of DOC concentration and isotopic fractionation due to incomplete oxidation. The development of an analytical method for accurate and sensitive DOC SIA is described in this study. METHODS: A high-temperature combustion (HTC) system improves upon traditional methods. A novel total organic carbon (TOC) system, specially designed for SIA, was coupled to an isotope ratio mass spectrometer. An integrated purge and trap technique (peak focusing), flexible injection volume (0.05-3 mL), favorable carrier gas flow, modified ash crucible, new design of combustion tube and optimized drying system were used to achieve the necessary performance. RESULTS: The system can reliably measure concentrations up to 1000 mgC/L. Compounds resistant to oxidation, such as barbituric acid, melamine and humic acid, were analyzed with recovery rates of 100 ± 1% proving complete oxidation. In this initial testing, the δ(13) C values of these compounds were determined with precision and trueness of ≤ 0.2‰ even with 3.5% salinity. Further tests with samples with low DOC concentrations resulted in LOQSIA method values of 0.5 mgC/L and 0.2 mgC/L for LOQSIA instr , considering an accuracy of ± 0.5‰ as acceptable. CONCLUSIONS: The novel HTC system coupled to an isotope ratio mass spectrometer resulted in significantly improved sensitivity. The system is suitable for salt-containing liquids and compounds that are resistant to oxidation, and it offers a large concentration range. A second paper (which follows this one in this issue) will present a more comprehensive assessment of the analytical performance with a broad set of solutions and real samples. This highly efficient TOC stable isotopic analyzer will probably open up new possibilities in biogeochemical carbon cycle research.

Microwave-assisted nonionic surfactant extraction of aliphatic hydrocarbons from petroleum source rock.

The extraction of aliphatic hydrocarbons from petroleum source rock using nonionic surfactants with the assistance of microwave was investigated and the conditions for maximum yield were determined. The results showed that the extraction temperatures and kinetic rates have significant effects on extraction yields of aliphatic hydrocarbons. The optimum temperature for microwave-assisted nonionic surfactant extraction of aliphatic hydrocarbons from petroleum source rock was 105°C. The optimum extraction time for the aliphatic hydrocarbons was at 50 min. Concentration of the nonionic surfactant solution and irradiation power had significant effect on the yields of aliphatic hydrocarbons. The yields of the analytes were much higher using microwave assisted nonionic surfactant extraction than with Soxhlet extraction. The recoveries of the n-alkanes and acyclic isoprenoid hydrocarbons for GC-MS analysis from the extractant nonionic surfactant solution by in-tube extraction (ITEX 2) with a TENAX TA adsorbent were found to be efficient. The results show that microwave-assisted nonionic surfactant extraction (MANSE) is a good and efficient green analytical preparatory technique for geochemical evaluation of petroleum source rock.

A strategy for the systematic development of a liquid chromatographic mass spectrometric screening method for polymer electrolyte membrane degradation products using isocratic and gradient phase optimized liquid chromatography.

Within the scope of research for target and non-target LC-MS/MS analysis of membrane degradation products of polymer electrolyte membrane fuel cells, a systematic method development for the separation of structurally similar compounds was performed by phase optimized liquid chromatography. Five different stationary phases with different selectivities were used. Isocratic separation for 4-hydroxybenzoic acid, isophthalic acid, terephthalic acid, 4-hydroxybenzaldehyde and 4-formylbenzoic acid was achieved on a C18 and a Phenyl phase. Using the PRISMA model the separation efficiency was optimized. This was achieved on a serially connected mixed stationary phase composed of 30 mm C18, 150 mm Phenyl and 60 mm C30. For the LC-MS screening of unknown degradation products from polymer electrolyte membranes in the product water of a fuel cell, a solvent gradient is mandatory for less polar or later eluting compounds. By means of 4-mercaptobenzoic acid it could be shown that a solvent gradient can be applied in order to elute later eluting compounds in a short time. The adaptability of this method for the qualitative analysis by target and non-target LC-MS/MS screening has been shown by means of 4-hydroxybenzoic acid. The combination of solvent gradient and isocratic conditions makes this approach attractive for the purpose of a screening method for known and unknown analytes in a water sample.

MTBE oxidation by conventional ozonation and the combination ozone/hydrogen peroxide: efficiency of the processes and bromate formation.

The present study investigates the oxidation of methyl tert-butyl ether (MTBE) by conventional ozonation and the advanced oxidation process (AOP) ozone/hydrogen peroxide under drinking water treatment conditions. The major degradation products identified were tert-butyl formate (TBF), tert-butyl alcohol (TBA), 2-methoxy-2-methyl propionaldehyde (MMP), acetone (AC), methyl acetate (MA), hydroxyisobutyraldehyde (HiBA), and formaldehyde (FA). The rate constants of the reaction of ozone and OH radicals with MTBE were found to be 0.14 and 1.9 x 10(9) M(-1) s(-1), respectively. The rate constants for the same oxidation processes were also measured for the degradation products TBF, MMP, MA, and HiBA (k(O3-TBF) = 0.78 M(-1) s(-1); k(OH-TBF) = 7.0 x 10(8) M(-1) s(-1); k(O3-MMP) = 5 M(-1) s(-1); k(OH-MMP) = 3 x 10(9) M(-1) s(-1), k(O3-MA) = 0.09 M(-1) s(-1), k(O3-HiBA) = 5 M(-1) s(-1); k(OH-HiBA) = 3 x 10(9) M(-1) s(-1)). Since all compounds reacted slowly with molecular ozone, only the degradation pathway of MTBE with OH radicals has been determined, including the formation of primary degradation products. In experiments performed with several natural waters, the efficiency of MTBE elimination and the formation of bromate as disinfection byproduct have been measured. With a bromide level of 50 microg/L, only 35-50% of MTBE could be eliminated by the AOP O3/H2O2 without exceeding the current drinking water standard of bromate (10 microg/L). The transient concentrations of MTBE and its primary degradation products were modeled using a combination of kinetic parameters (degradation product distribution and rate constants) together with the ozone and OH radical concentration and were in good agreement with the experimental results.

Identification of oxidized TNT metabolites in soil samples of a former ammunition plant.

Water extracts of soil samples of the former ammunition plant "Tanne" near Clausthal-Zellerfeld, Lower Saxony, Germany, were investigated for highly polar oxidized 2,4,6-trinitrotoluene (TNT) metabolites. 0.4 to 9.0 mg/kg dry soil 2,4,6-trinitrobenzoic acid (TNBA) and 5.8 to 544 mg/kg dry soil 2-amino-4,6-dinitrobenzoic acid (2-ADNBA) were found. In addition to the oxidized metabolites, TNT, 4- and 2-aminodinitrotoluene (4- and 2-ADNT), and 2,4-dinitrotoluene (2,4-DNT) were extractable with water. Most interestingly, in one sample, 2-ADNBA represented the main contaminant. The origin of the oxidized nitroaromatics is unknown at this time. They might be generated chemically or photochemically. Furthermore, a biological synthesis seems possible.

Highly polar metabolites of nitroaromatic compounds in ammunition wastewater.

Transformation processes of nitroaromatic compounds (NAC) are described that lead to polar and highly hydrophilic metabolites in ammunition waste water. For the identification of proposed metabolites, several analytical methods for the separation and determination of these compounds were developed and applied to the investigation of a former ammunition plant and its surroundings. The compound classes which were investigated include nitro and amino substituted toluenesulfonic acids, nitrobenzoic acids, aminobenzoic acids and nitromethylphenols. Method development was usually done with high performance liquid chromatography and ion pairing or ion suppression techniques. In the leachate water of a disposal site of a former ammunition plant in Stadtallendorf, Germany, 15 compounds from the above-mentioned categories have been found to be present in concentrations up to a few hundred microgram/L. The presence of so many polar compounds suggest that current and future surveying programs should include representative analytes of the investigated categories.

Gas chromatographic determination of aromatic amines in water samples after solid-phase extraction and derivatization with iodine. I. Derivatization.

A new method for the selective determination of aromatic amines is presented, which is based on the solid-phase extraction at pH 9 and subsequent derivatization of the analytes to the corresponding iodobenzenes. These can selectively and sensitively be determined with gas chromatography and electron-capture detection. Separation of at least 30 compounds in a single chromatographic run in 30 min is possible. With this method, 56 aromatic amines were investigated, and only in six cases no derivatives were obtained. Limits of quantitation were between 0.5 and 8 micrograms 1(-1), but may still be lowered with higher sample volumes or different injection techniques. The application to water samples revealed the suitability for the investigation of ground, leachate and wastewater.

Gas chromatographic determination of aromatic amines in water samples after solid-phase extraction and derivatization with iodine. II. Enrichment.

A procedure for the enrichment of aromatic amines via solid-phase extraction was developed. A HR-P phase based on styrene-divinylbenzene was used for the investigations, generally followed by derivatization with iodine and determination via GC-ECD. The recoveries of 53 aromatic amines in a drinking water matrix at pH 9 were determined. Most anilines showed relative recoveries between 80-120% with relative standard deviations of< or = 5% at concentration levels between 10 and 20 micrograms 1(-1). The comparison with a wastewater matrix led to similar results. The enrichment procedure was applied to real samples, e.g., samples of ammunition wastewater.