Biodegradable polymers boost reproduction in the earthworm Eisenia fetida.

Microplastic contamination in soil has become a global environmental threat as it adversely affects terrestrial organisms like earthworms as well as soil properties. Especially biodegradable polymers have recently been used as an alternative to conventional polymer types, although their impact remains poorly understood. Thus, we studied the effect of conventional (polystyrene: PS, polyethylene terephthalate: PET, polypropylene: PP) versus aliphatic polyesters classified as biodegradable polymers (poly-(l-lactide): PLLA, polycaprolactone: PCL) on the earthworm Eisenia fetida and soil properties (pH and cation exchange capacity). We addressed direct effects on the weight gain and reproductive success of E. fetida, and indirect effects, like changes in the gut microbial composition as well as the production of short-chain fatty acids by the gut microbiota. Earthworms were exposed for eight weeks in an artificial soil amended with two environmentally relevant concentrations (1 % and 2.5 % (w/w)) of the different microplastic types. PLLA and PCL boosted the number of cocoons produced by 135 % and 54 %, respectively. Additionally, exposure to these two polymers increased number of hatched juveniles, changed gut microbial beta-diversity, and increased the production of the short chain fatty acid lactate compared to the control treatments. Interestingly, we also found a positive effect of PP on the earthworm's bodyweight and reproductive success. The interaction of microplastic and earthworms decreased soil pH by about 1.5 units in the presence of PLLA and PCL. No polymer effect on the cation exchange capacity of soil was found. In general, neither the presence of conventional nor biodegradable polymers had any adverse effects on any of the studied endpoints. Our results suggest that the effects of microplastic highly depend on the polymer type, and that the degradation of biodegradable polymers might be enhanced in the gut of earthworms, which implies that they may use biodegradable polymers as a potential carbon source.

Correlative Imaging of the Rhizosphere─A Multimethod Workflow for Targeted Mapping of Chemical Gradients.

Examining in situ processes in the soil rhizosphere requires spatial information on physical and chemical properties under undisturbed conditions. We developed a correlative imaging workflow for targeted sampling of roots in their three-dimensional (3D) context and assessed the imprint of roots on chemical properties of the root-soil contact zone at micrometer to millimeter scale. Maize (Zea mays) was grown in 15N-labeled soil columns and pulse-labeled with 13CO2 to visualize the spatial distribution of carbon inputs and nitrogen uptake together with the redistribution of other elements. Soil columns were scanned by X-ray computed tomography (X-ray CT) at low resolution (45 µm) to enable image-guided subsampling of specific root segments. Resin-embedded subsamples were then analyzed by X-ray CT at high resolution (10 µm) for their 3D structure and chemical gradients around roots using micro-X-ray fluorescence spectroscopy (µXRF), nanoscale secondary ion mass spectrometry (NanoSIMS), and laser-ablation isotope ratio mass spectrometry (LA-IRMS). Concentration gradients, particularly of calcium and sulfur, with different spatial extents could be identified by µXRF. NanoSIMS and LA-IRMS detected the release of 13C into soil up to a distance of 100 µm from the root surface, whereas 15N accumulated preferentially in the root cells. We conclude that combining targeted sampling of the soil-root system and correlative microscopy opens new avenues for unraveling rhizosphere processes in situ.

Silicon as a potential limiting factor for phosphorus availability in paddy soils.

Rice cultivation requires high amounts of phosphorus (P). However, significant amounts of P fertilizer additions may be retained by iron (Fe) oxides and are thus unavailable for plants. At the same time, rice cultivation has a high demand for silicic acid (Si), reducing Si availability after short duration of rice cultivation. By studying a paddy chronosequence with rice cultivation up to 2000 years, we show that Si limitation, observed as early as a few decades of rice cultivation, is limiting P availability along the paddy soils chronosequence. Using near edge X-ray absorption fine structure spectroscopy (NEXAFS) in a scanning transmission (soft) X-ray microscope (STXM) we show release of available P was linked to a Si-induced change in speciation of Fe-phases in soil particles and competition of Si with P for binding sites. Hence, low Si availability is limiting P availability in paddy soils. We propose that proper management of Si availability is a promising tool to improve the P supply of paddy plants.

Bioturbation by black soldier fly larvae-Rapid soil formation with burial of ceramic artifacts.

Bioturbation involves the incorporation of residues from the surface soil into the subsoil; however, common small soil 'bioengineers', such as earthworms or termites, are unlikely to transport human artifacts to deeper soil horizons. However, such artifacts occur in the deeper soil horizons within Amazonian Anthrosols (Terra Preta). Here we test the assumption that such tasks could be carried out by fly larvae, which could thus play a crucial role in waste decomposition and associated soil mixing under tropical conditions. We performed two greenhouse experiments with sandy substrate covered with layers of organic waste, ceramic fragments, and black soldier fly larvae (BSFL) (Hermetia illucens (L.) (Dipt.: Stratiomyidae)). We used in-situ images to assess the rate of bioturbation by BSFL, and then designed our main study to observe waste dissipation (reduction of organic carbon and phosphorus contents from waste model trials with and without charcoal) as related to larval-induced changes in soil properties. We found that the bioturbation of macroinvertebrates like BSFL was able to bury even large (> 5 cm) ceramic fragments within hours, which coincided with high soil growth rates (0.5 cm h-1). The sandy soil was subsequently heavily enriched with organic matter and phosphorus originating from organic waste. We conclude that BSFL, and possibly other fly species, are important, previously overlooked soil 'bioengineers', which may even contribute to the burial of artifacts in Anthrosols and other terrestrial waste dumps.

Evidence for signatures of ancient microbial life in paleosols.

Loess-paleosol sequences are terrestrial archives of past climate change. They may host traces of ancient microbial life, but little information is available on the recovery of microbial biomarkers from such deposits. We hypothesized that microbial communities in soil horizons up to an age of 127 kyr carry information related to past environments. We extracted DNA from a loess-paleosol sequence near Toshan, Northern Iran, with 26 m thick deposits showing different degrees of soil development, performed quantitative PCR and 16S rRNA gene amplicon sequencing. Periods of soil formation archived within the loess sediment led to higher diversity and bacterial abundance in the paleosol horizons. Community composition fluctuated over the loess-paleosol sequence and was mainly correlated with age and depth, (ADONIS R2 < 0.14, P ≤ 0.002), while responses to paleosol soil traits were weaker. Phyla like Bacteriodetes, Proteobacteria or Acidobacteria were more prevalent in paleosol horizons characterized by intense soil formation, while weakly developed paleosols or loess horizons hosted a higher percentage and diversity of Actinobacteria. Taken together, our findings indicate that the microbial community in loess-paleosol sequences carries signatures of earlier environmental conditions that are preserved until today.

Spatial Microanalysis of Natural 13C/12C Abundance in Environmental Samples Using Laser Ablation-Isotope Ratio Mass Spectrometry.

The stable 13C/12C isotope composition usually varies among different organic materials due to isotope fractionation during biochemical synthesis and degradation processes. Here, we introduce a novel laser ablation-isotope ratio mass spectrometry (LA-IRMS) methodology that allows highly resolved spatial analysis of carbon isotope signatures in solid samples down to a spatial resolution of 10 µm. The presented instrumental setup includes in-house-designed exchangeable ablation cells (3.8 and 0.4 mL, respectively) and an improved sample gas transfer, which allow accurate δ13C measurements of an acryl plate standard down to 0.6 and 0.4 ng of ablated carbon, respectively (standard deviation 0.25‰). Initial testing on plant and soil samples confirmed that microheterogeneity of their natural 13C/12C abundance can now be mapped at a spatial resolution down to 10 µm. The respective δ13C values in soils with C3/C4 crop sequence history varied by up to 14‰ across a distance of less than 100 µm in soil aggregates, while being partly sorted along rhizosphere gradients of <300 µm from Miscanthus plant roots into the surrounding soil. These very first demonstrations point to the appearance of very small metabolic hotspots originating from different natural isotope discrimination processes, now traceable via LA-IRMS.

Inland navigation: PAH inventories in soil and vegetation after EU fuel regulation 2009/30/EC.

In January 2011, fuel quality in inland water vessels was changed by EU regulation 2009/30/EC, aiming at improving air quality along waterways. We hypothesized that the implementation of this regulation both lowered the total deposition of polycyclic aromatic hydrocarbons (PAHs) and changed their composition in river valleys. We analyzed parent-, alkylated- and thio-PAHs in soil and vine leaves, at two waterways (Rhine and Moselle, Germany), as well as in one ship-free reference area (Ahr, Germany). Samples were taken annually (2010-2013) in transects perpendicular to the rivers. We did not find any relation of PAH concentration and composition on vine leaves to inland navigation, likely because atmospheric exchange processes distorted ship-specific accumulation patterns. We did find, however, an accumulation of ship-borne PAHs in topsoil near the waterways (1543±788 and 581±252ngg-1 at Moselle and Rhine, respectively), leading to larger PAH concentrations at the Moselle Valley than at the reference area (535±404ngg-1) prior to EU fuel regulation. After fuel regulation, the PAH concentrations decreased in topsoils of the Moselle and Rhine Valley by 35±9 and 62±28%, respectively. These changes were accompanied by increasing proportions of dibenzothiophene (DBT) and low molecular weight PAHs. Both, changes in PAH concentrations and composition were traceable within 200 and 350m distance to the river front of Moselle and Rhine, respectively, and likely favored by erosion of topsoil in vineyards. We conclude that the EU regulation was effective in improving soil and thus also air quality within only three years. The impact was greater and spatially more relevant at the Rhine, which may be attributed to the larger traffic volume of inland navigation.

Steroid Biomarkers Revisited - Improved Source Identification of Faecal Remains in Archaeological Soil Material.

Steroids are used as faecal markers in environmental and in archaeological studies, because they provide insights into ancient agricultural practices and the former presence of animals. Up to now, steroid analyses could only identify and distinguish between herbivore, pig, and human faecal matter and their residues in soils and sediments. We hypothesized that a finer differentiation between faeces of different livestock animals could be achieved when the analyses of several steroids is combined (Δ5-sterols, 5α-stanols, 5ß-stanols, epi-5ß-stanols, stanones, and bile acids). We therefore reviewed the existing literature on various faecal steroids from livestock and humans and analysed faeces from old livestock breed (cattle, horse, donkey, sheep, goat, goose, and pig) and humans. Additionally, we performed steroid analyses on soil material of four different archaeological periods (sites located in the Lower Rhine Basin, Western Germany, dating to the Linearbandkeramik, Urnfield Period / Bronze Age, Iron Age, Roman Age) with known or supposed faecal inputs. By means of already established and newly applied steroid ratios of the analysed faeces together with results from the literature, all considered livestock faeces, except sheep and cattle, could be distinguished on the basis of their steroid signatures. Most remarkably was the identification of horse faeces (via the ratio: epi-5ß-stigmastanol: 5ß-stigmastanol + epicoprostanol: coprostanol; together with the presence of chenodeoxycholic acid) and a successful differentiation between goat (with chenodeoxycholic acid) and sheep/cattle faeces (without chenodeoxycholic acid). The steroid analysis of archaeological soil material confirmed the supposed faecal inputs, even if these inputs had occurred several thousand years ago.

Organic fertilization and sufficient nutrient status in prehistoric agriculture?--Indications from multi-proxy analyses of archaeological topsoil relicts.

Neolithic and Bronze Age topsoil relicts revealed enhanced extractable phosphorus (P) and plant available inorganic P fractions, thus raising the question whether there was targeted soil amelioration in prehistoric times. This study aimed (i) at assessing the overall nutrient status and the soil organic matter content of these arable topsoil relicts, and (ii) at tracing ancient soil fertilizing practices by respective stable isotope and biomarker analyses. Prehistoric arable topsoils were preserved in archaeological pit fillings, whereas adjacent subsoils served as controls. One Early Weichselian humic zone represented the soil status before the introduction of agriculture. Recent topsoils served as an additional reference. The applied multi-proxy approach comprised total P and micronutrient contents, stable N isotope ratios, amino acid, steroid, and black carbon analyses as well as soil color measurements. Total contents of P and selected micronutrients (I, Cu, Mn, Mo, Se, Zn) of the arable soil relicts were above the limits for which nutrient deficiencies could be assumed. All pit fillings exhibited elevated δ15N values close to those of recent topsoils (δ15N>6 to 7‰), giving first hints for prehistoric organic N-input. Ancient legume cultivation as a potential source for N input could not be verified by means of amino acid analysis. In contrast, bile acids as markers for faecal input exhibited larger concentrations in the pit fillings compared with the reference and control soils indicating faeces (i.e. manure) input to Neolithic arable topsoils. Also black carbon contents were elevated, amounting up to 38% of soil organic carbon, therewith explaining the dark soil color in the pit fillings and pointing to inputs of burned biomass. The combination of different geochemical analyses revealed a sufficient nutrient status of prehistoric arable soils, as well as signs of amelioration (inputs of organic material like charcoal and faeces-containing manure).

Black carbon accrual during 2000 years of paddy-rice and non-paddy cropping in the Yangtze River Delta, China.

Rice straw burning has accompanied paddy management for millennia, introducing black carbon (BC) into soil as the residue of incomplete combustion. This study examined the contribution of BC to soil organic matter and the rate at which it accumulates in paddy soils as a result of prolonged paddy management. Soil depth profiles were sampled along a chronosequence of 0-2000 years of rice-wheat rotation systems and adjacent non-paddy systems (50-700 years) in the Bay of Hangzhou (Zhejiang province, China). The soil BC content and its degree of condensation were assessed using benzene-polycarboxylic acids (BPCAs) as geochemical markers. The results showed that despite regular long term BC input, BC only contributed 7-11% of total soil organic carbon (SOC) in the topsoil horizons. Nevertheless, along with SOC, paddy soils accumulated BC with increasing duration of management until 297 years to reach a steady-state of 13 t BC ha(-1). This was 1.8 times more than in non-paddy soils. The fate of BC in paddy soils (0-1 m) could be modeled revealing an average annual input of 44 kg ha(-1) yr(-1), and a mean residence time of 303 years. The subsoils contributed at least 50% to overall BC stocks, which likely derived from periods prior to land embankment and episodic burial of ancient topsoil, as also indicated by BPCA pattern changes. We conclude that there is a significant but limited accumulation of C in charred forms upon prolonged paddy management. The final contribution of BC to total SOC in paddy soils was similar to that in other aerobic ecosystems of the world.

Early and middle holocene hunter-gatherer occupations in western Amazonia: the hidden shell middens.

We report on previously unknown early archaeological sites in the Bolivian lowlands, demonstrating for the first time early and middle Holocene human presence in western Amazonia. Multidisciplinary research in forest islands situated in seasonally-inundated savannahs has revealed stratified shell middens produced by human foragers as early as 10,000 years ago, making them the oldest archaeological sites in the region. The absence of stone resources and partial burial by recent alluvial sediments has meant that these kinds of deposits have, until now, remained unidentified. We conducted core sampling, archaeological excavations and an interdisciplinary study of the stratigraphy and recovered materials from three shell midden mounds. Based on multiple lines of evidence, including radiocarbon dating, sedimentary proxies (elements, steroids and black carbon), micromorphology and faunal analysis, we demonstrate the anthropogenic origin and antiquity of these sites. In a tropical and geomorphologically active landscape often considered challenging both for early human occupation and for the preservation of hunter-gatherer sites, the newly discovered shell middens provide evidence for early to middle Holocene occupation and illustrate the potential for identifying and interpreting early open-air archaeological sites in western Amazonia. The existence of early hunter-gatherer sites in the Bolivian lowlands sheds new light on the region's past and offers a new context within which the late Holocene "Earthmovers" of the Llanos de Moxos could have emerged.