217 000-year-old DNA sequences of green sulfur bacteria in Mediterranean sapropels and their implications for the reconstruction of the paleoenvironment

Deep-sea sediments of the eastern Mediterranean harbour a series of dark, organic carbon-rich layers, so-called sapropels. Within these layers, the carotenoid isorenieratene was detected. Since it is specific for the obligately anaerobic phototrophic green sulfur bacteria, the presence of isorenieratene may suggest that extended water column anoxia occurred in the ancient Mediterranean Sea during periods of sapropel formation. Only three carotenoids (isorenieratene, beta-isorenieratene and chlorobactene) are typical for green sulfur bacteria and thus do not permit to differentiate between the approximately 80 known phylotypes. In order to reconstruct the paleoecological conditions in more detail, we searched for fossil 16S rRNA gene sequences of green sulfur bacteria employing ancient DNA methodology. 540 bp-long fossil sequences could indeed be amplified from up to 217 000-year-old sapropels. In addition, such sequences were also recovered from carbon-lean intermediate sediment layers deposited during times of an entirely oxic water column. Unexpectedly, however, all the recovered 16S rRNA gene sequences grouped with freshwater or brackish, rather than truly marine, types of green sulfur bacteria. It is therefore feasible that the molecular remains of green sulfur bacteria originated from populations which thrived in adjacent freshwater or estuarine coastal environments rather than from an indigenous pelagic population.     

Ancient DNA complements microfossil record in deep-sea subsurface sediments

Deep-sea subsurface sediments are the most important archives of marine biodiversity. Until now, these archives were studied mainly using the microfossil record, disregarding large amounts of DNA accumulated on the deep-sea floor. Accessing ancient DNA (aDNA) molecules preserved down-core would offer unique insights into the history of marine biodiversity, including both fossilized and non-fossilized taxa. Here, we recover aDNA of eukaryotic origin across four cores collected at abyssal depths in the South Atlantic, in up to 32.5 thousand-year-old sediment layers. Our study focuses on Foraminifera and Radiolaria, two major groups of marine microfossils also comprising diverse non-fossilized taxa. We describe their assemblages in down-core sediment layers applying both micropalaeontological and environmental DNA sequencing approaches. Short fragments of the foraminiferal and radiolarian small subunit rRNA gene recovered from sedimentary DNA extracts provide evidence that eukaryotic aDNA is preserved in deep-sea sediments encompassing the last glacial maximum. Most aDNA were assigned to non-fossilized taxa that also dominate in molecular studies of modern environments. Our study reveals the potential of aDNA to better document the evolution of past marine ecosystems and opens new horizons for the development of deep-sea palaeogenomics.     

Eukaryotic diversity in late Pleistocene marine sediments around a shallow methane hydrate deposit in the Japan Sea

Marine sediments contain eukaryotic DNA deposited from overlying water columns. However, a large proportion of deposited eukaryotic DNA is aerobically biodegraded in shallow marine sediments. Cold seep sediments are often anaerobic near the sediment–water interface, so eukaryotic DNA in such sediments is expected to be preserved. We investigated deeply buried marine sediments in the Japan Sea, where a methane hydrate deposit is associated with cold seeps. Quantitative PCR analysis revealed the reproducible recovery of eukaryotic DNA in marine sediments at depths up to 31.0 m in the vicinity of the methane hydrate deposit. In contrast, the reproducible recovery of eukaryotic DNA was limited to a shallow depth (8.3 m) in marine sediments not adjacent to the methane hydrate deposit in the same area. Pyrosequencing of an 18S rRNA gene variable region generated 1,276–3,307 reads per sample, which was sufficient to cover the biodiversity based on rarefaction curves. Phylogenetic analysis revealed that most of the eukaryotic DNA originated from radiolarian genera of the class Chaunacanthida, which have SrSO₄ skeletons, the sea grass genus Zostera, and the seaweed genus Sargassum. Eukaryotic DNA originating from other planktonic fauna and land plants was also detected. Diatom sequences closely related to Thalassiosira spp., indicative of cold climates, were obtained from sediments deposited during the last glacial period (MIS‐2). Plant sequences of the genera Alnus, Micromonas, and Ulmus were found in sediments deposited during the warm interstadial period (MIS‐3). These results suggest the long‐term persistence of eukaryotic DNA from terrestrial and aquatic sources in marine sediments associated with cold seeps, and that the genetic information from eukaryotic DNA from deeply buried marine sediments associated with cold seeps can be used to reconstruct environments and ecosystems from the past.     

Preservation, origin and genetic imprint of extracellular DNA in permanently anoxic deep-sea sediments

Molecular approaches that target the total DNA pool recovered from permanently anoxic marine ecosystems have revealed an extraordinary diversity of prokaryotes and unicellular eukaryotes. However, the presence of gene sequences contained within the extracellular DNA pool is still largely neglected. We have investigated the preservation, origin and genetic imprint of extracellular DNA recovered from permanently anoxic deep-sea sediments of the Black Sea. Despite high DNase activities, huge amounts of total extracellular DNA were found in both the surface and subsurface sediment layers, suggesting reduced availability of the extracellular DNA pool to nuclease degradation. The reduced degradation of the total extracellular DNA was confirmed by its low decay rate and the high accumulation in the deeper sediment layers. The copy numbers of 16S and 18S rDNA contained within the extracellular DNA pool in both the surface and subsurface sediment layers was very high, indicating that permanently anoxic sediments of the deep Black Sea are hot spots of preserved extracellular gene sequences. The extracellular DNA recovered from these sediment layers also contained highly diversified 18S rDNA sequences. These were not only representative of the major protistan lineages, but also of new very divergent lineages, branching as independent clades at the base of the tree. Our findings indicate that the extracellular DNA pool is a major archive of present/past eukaryotic gene sequences, and they highlight the importance of integrating molecular cell-oriented approaches with molecular analyses of the extracellular DNA pool, for a better assessment of microbial diversity and temporal changes in marine benthic ecosystems.     

Functional exoenzymes as indicators of metabolically active bacteria in 124,000-year-Old sapropel layers of the eastern Mediterranean Sea

Hydrolytic exoenzymes as indicators of metabolically active bacteria were investigated in four consecutive sapropel layers collected from bathyal sediments of the eastern Mediterranean Sea. For comparison, the organic carbon-poor layers between the sapropels, sediment from the anoxic Urania basin, and sediments of intertidal mud flats of the German Wadden Sea were also analyzed. The sapropel layers contained up to 1.5. 10(8) bacterial cells cm(-3), whereas cell numbers in the intermediate layers were lower by a factor of 10. In sapropels, the determination of exoenzyme activity with fluorescently labeled substrate analogues was impaired by the strong adsorption of up to 97% of the enzymatically liberated fluorophores (4-methylumbelliferone [MUF] and 7-amino-4-methylcoumarin [MCA]) to the sediment particles. Because all established methods for the extraction of adsorbed fluorophores proved to be inadequate for sapropel sediments, we introduce a correction method which is based on the measurement of equilibrium adsorption isotherms for both compounds. Using this new approach, high activities of aminopeptidase and alkaline phosphatase were detected even in a 124,000-year-old sapropel layer, whereas the activity of beta-glucosidase was low in all layers. So far, it had been assumed that the organic matter which constitutes the sapropels is highly refractory. The high potential activities of bacterial exoenzymes indicate that bacteria in Mediterranean sapropels are metabolically active and utilize part of the subfossil kerogen. Since a high adsorption capacity was determined not only for the low-molecular-weight compounds MUF and MCA but also for DNA, the extraordinarily strong adsorption of structurally different substrates to the sapropel matrix appears to be the major reason for the long-term preservation of biodegradable carbon in this environment.     

Widespread distribution and high abundance of Rhizobium radiobacter within Mediterranean subsurface sediments

Eastern Mediterranean sediments are characterized by the occurrence of distinct, organic-rich layers, called sapropels. These harbour elevated microbial numbers in comparison with adjacent carbon-lean intermediate layers. A recently obtained culture collection from these sediments was composed of 20% of strains closely related to Rhizobium radiobacter, formerly classified as Agrobacterium tumefaciens. To prove and quantify the in situ abundance of R. radiobacter, a highly specific quantitative polymerase chain reaction (PCR) protocol was developed. To convert quantification results into cell numbers, the copy number of rrn operons per genome was determined. Southern hybridization showed that our isolates contained four operons. Finally, quantitative PCR was applied to 45 sediment samples obtained across the eastern Mediterranean. Rhizobium radiobacter was present in 38 of 45 samples indicating an almost ubiquitous distribution. In total, 25-40 000 cells per gram of sediment were detected, corresponding to 0.001-5.1% of the bacterial cells. In general, the relative and absolute abundance of R. radiobacter increased with depth and was higher in sapropels than in intermediate layers. This indicates that R. radiobacter forms an active population in up to 200 000 years old sapropels. The present study shows for the first time that a cultivated subsurface bacterium is highly abundant in this environment.     

Functional Exoenzymes as Indicators of Metabolically Active Bacteria in 124,000-Year-Old Sapropel Layers of the Eastern Mediterranean Sea

Hydrolytic exoenzymes as indicators of metabolically active bacteria were investigated in four consecutive sapropel layers collected from bathyal sediments of the eastern Mediterranean Sea. For comparison, the organic carbon-poor layers between the sapropels, sediment from the anoxic Urania basin, and sediments of intertidal mud flats of the German Wadden Sea were also analyzed. The sapropel layers contained up to 1.5 · 10⁸ bacterial cells cm⁻³, whereas cell numbers in the intermediate layers were lower by a factor of 10. In sapropels, the determination of exoenzyme activity with fluorescently labeled substrate analogues was impaired by the strong adsorption of up to 97% of the enzymatically liberated fluorophores (4-methylumbelliferone [MUF] and 7-amino-4-methylcoumarin [MCA]) to the sediment particles. Because all established methods for the extraction of adsorbed fluorophores proved to be inadequate for sapropel sediments, we introduce a correction method which is based on the measurement of equilibrium adsorption isotherms for both compounds. Using this new approach, high activities of aminopeptidase and alkaline phosphatase were detected even in a 124,000-year-old sapropel layer, whereas the activity of β-glucosidase was low in all layers. So far, it had been assumed that the organic matter which constitutes the sapropels is highly refractory. The high potential activities of bacterial exoenzymes indicate that bacteria in Mediterranean sapropels are metabolically active and utilize part of the subfossil kerogen. Since a high adsorption capacity was determined not only for the low-molecular-weight compounds MUF and MCA but also for DNA, the extraordinarily strong adsorption of structurally different substrates to the sapropel matrix appears to be the major reason for the long-term preservation of biodegradable carbon in this environment.     

Spatial productivity variations during formation of sapropels S5 and S6 in the Mediterranean Sea: evidence from Ba contents

We investigated five time-equivalent core sections (180-110 kyr BP) from the Balearic Sea (Menorca Rise), the easternmost Levantine Basin and southwest, south, and southeast of Crete to reconstruct spatial patterns of productivity during deposition of sapropels S5 and S6 in the Mediterranean Sea. Our indicators are Ba, total organic carbon and carbonate contents. We found no indications of Ba remobilization within the investigated core intervals, and used the accumulation rate of biogenic Ba to compute paleoproductivity. Maximum surface water productivity (up to 350 g C/m²/yr) was found during deposition of S5 (isotope stage 5e) but pronounced spatial variability is evident. Coeval sediment intervals in the Balearic Sea show very little productivity change, suggesting that chemical and biological environments in the eastern and western Mediterranean basins were decoupled in this interval. We interpret the spatial variability as the result of two different modes of nutrient delivery to the photic zone: river-derived nutrient input and shoaling of the pycnocline/nutricline to the photic zone. The productivity increase during the formation of S6 was moderate compared to S5 and had a less marked spatial variability within the study area of the eastern Mediterranean Sea. Given that S6 formed during a glacial interval, glacial boundary conditions such as high wind stress and/or cooler surface water temperatures apparently favored lateral and vertical mixing and prevented the development of the spatial gradients within the Eastern Mediterranean Sea (EMS) observed for S5. A non-sapropel sediment interval with elevated Ba content and depleted ¹⁸O/¹⁶O ratios in planktonic foraminifer calcite was detected between S6 and S5 that corresponds to the weak northern hemisphere insolation maximum at 150 kyr. At this time, productivity apparently increased up to five times over surrounding intervals, but abundant benthic fauna show that the deep water remained oxic. Following our interpretation, the interval denotes a failed sapropel, when a weaker monsoon did not force the EMS into permanent stratification. The comparison of interglacial and glacial sapropels illustrates the relevance of climatic boundary conditions in the northern catchment in determining the facies and spatial variability of sapropels within the EMS.     

Ancient DNA sheds new light on the Svalbard foraminiferal fossil record of the last millennium

Recent palaeogenetic studies have demonstrated the occurrence of preserved ancient DNA (aDNA) in various types of fossilised material. Environmental aDNA sequences assigned to modern species have been recovered from marine sediments dating to the Pleistocene. However, the match between the aDNA and the fossil record still needs to be evaluated for the environmental DNA approaches to be fully exploited. Here, we focus on foraminifera in sediments up to one thousand years old retrieved from the Hornsund fjord (Svalbard). We compared the diversity of foraminiferal microfossil assemblages with the diversity of aDNA sequenced from subsurface sediment samples using both cloning and high‐throughput sequencing (HTS). Our study shows that 57% of the species archived in the fossil record were also detected in the aDNA data. However, the relative abundance of aDNA sequence reads and fossil specimens differed considerably. We also found a limited match between the stratigraphic occurrence of some fossil species and their aDNA sequences, especially in the case of rare taxa. The aDNA data comprised a high proportion of non‐fossilised monothalamous species, which are known to dominate in modern foraminiferal communities of the Svalbard region. Our results confirm the relevance of HTS for studying past micro‐eukaryotic diversity and provide insight into its ability to reflect fossil assemblages. Palaeogenetic studies including aDNA analyses of non‐fossilised groups expand the range of palaeoceanographical proxies and therefore may increase the accuracy of palaeoenvironmental reconstructions.     

山东半岛南部近岸海域晚第四纪以来有孔虫和介形类化石群落分布特征及古环境演化

在山东半岛南部近岸海域QDZ03孔16.2m的沉积记录中,通过有孔虫和介形类化石群落的定量分析,探讨了研究区晚第四纪氧同位素(MIS)4期以来古沉积环境演变。在距今6万年前后开始的黄海海退期(相当于MIS4期),发育了有多次沉积间断的第1层(孔深16.2~12.0m)陆相地层。进入玉木亚间冰期(相当于MIS3)后,由于海平面的震荡上升,发育了第2层—第5层的滨岸弱海相—陆相沉积层,其中第5层上部可能包含部分MIS2期的产物,而MIS2期绝大部分沉积地层缺失。在距今12 000—11 000年前,海水逐渐由黄海入侵到青岛地区,发育了第6层下段(孔深9.1—8.6m)的海陆过渡相沉积层;在全新世最大洪泛面MFS来临之前(距今约11.0—9.0ka期间),海平面进一步上升,钻孔中发育了第6层中段(孔深8.6—7.7m)的滨岸相沉积层。随着距今6 000年前南黄海最大海泛面出现,后降至现今水平,并基本保持稳定状态,在黄河、近岸短源河流以及海岸带基岩侵蚀物等物质联合贡献下完成了南黄海中部泥质沉积体在近岸的延伸(7.7m以上),其中顶部约4.8m以上粒度粗化及与之相关的较多含量胶结壳有孔虫化石的出现,可能与约3 000—2 000年以来海岸带地区人类活动的加剧有关。     

黄海海域海洋沉积物细菌多样性分析

【背景】海洋独特的环境造就了海洋生物的多样性,海洋沉积物中细菌对海洋环境具有至关重要的作用。【目的】研究陆地土壤和海洋沉积物间细菌群落相似性和差异性,以便更好地认识海洋细菌多样性,深入了解沉积物细菌在海洋环境中的潜在作用。【方法】从中国黄海海域及大连市大黑山脚下分别采集样品,以陆地土壤为对照,采用16SrRNA基因高通量测序技术分析海洋沉积物的细菌群落结构。【结果】海洋沉积物样品中芽孢杆菌纲(Bacilli)、鞘氨醇单胞菌属(Sphingomonas)和芽孢杆菌属(Bacillus)丰度高于陆地土壤样品;海洋沉积物中亚硝化单胞菌(unculturedbacterium f. Nitrosomonadaceae)和厌氧绳菌(uncultured bacterium f. Anaerolineaceae)丰度虽低于陆地土壤,但丰度值也均高于1%;样品分类学统计显示酸杆菌门(Acidobacteria)在海洋沉积物和陆地土壤样品中的序列丰度比例都较大,鞘氨醇单胞菌属(Sphingomonas)在海洋沉积物样品中的序列丰度大于陆地土壤样品。【结论】海洋沉积物细菌多样性可作为海洋环境恢复情况的重...     

The influence of climate and sea-level change on the Holocene evolution of a Mediterranean coastal lagoon: Evidence from ostracod palaeoecology and geochemistry

Coastal lagoons provide an excellent basis for the study of processes controlling the evolution of a coastal zone. We examine the relative importance of these processes during the middle to late Holocene through a study of an 8.5 meter-long sediment record from the Albufera de Valencia (Spain). We combine sedimentological analyses with investigations into the palaeoecology, taphonomy and geochemistry (Mg/Ca, Sr/Ca, δ¹⁸O and δ¹³C) of ostracod valves in order to assess the effects of sea-level changes, storm events and effective moisture on the evolution of a Western Mediterranean coastal wetland. The late Pleistocene sediments represent a subaerial environment, which was followed by a hiatus in deposition. The first Holocene unit (～8700–7500 calendar yr. BP) is composed of typical lagoon-barrier and backshore sediments, deposited when seawater intruded into the lake and the climate was arid. The upper part of the sequence (between 7500 and 3400 yr.) is characterized by two sedimentary units, which correspond to Holocene progradation phases and humid climate associated with an increased freshwater influx to the lake accompanied by several high-energy events (palaeostorms). Overall, the record shows that an arid climate prevailed in the western Mediterranean area between 8400 and 7600 yr. The main marine transgression and accompanying progradational phases occurred between 7000 and 3400 yr., which is confirmed by other studies of coastal evolution along the Mediterranean coast. The multiproxy reconstructions demonstrate that controls on sedimentation and palaeoecology in this Mediterranean coastal lagoon were complex.     

Quantification of microbial communities in near-surface and deeply buried marine sediments on the Peru continental margin using real-time PCR

Deeply buried marine sediments harbour a large fraction of all prokaryotes on Earth but it is still unknown which phylogenetic and physiological microbial groups dominate the deep biosphere. In this study real-time PCR allowed a comparative quantitative microbial community analysis in near-surface and deeply buried marine sediments from the Peru continental margin. The 16S rRNA gene copy numbers of prokaryotes and Bacteria were almost identical with a maximum of 10(8)-10(10) copies cm(-3) in the near-surface sediments. Archaea exhibited one to three orders of magnitude lower 16S rRNA gene copy numbers. The 18S rRNA gene of Eukarya was always at least three orders of magnitude less abundant than the 16S rRNA gene of prokaryotes. The 16S rRNA gene of the Fe(III)- and Mn(IV)-reducing bacterial family Geobacteraceae and the dissimilatory (bi)sulfite reductase gene (dsrA) of sulfate-reducing prokaryotes were abundant with 10(6)-10(8) copies cm(-3) in near-surface sediments but showed lower numbers and an irregular distribution in the deep sediments. The copy numbers of all genes decreased with sediment depth exponentially. The depth gradients were steeper for the gene copy numbers than for numbers of total prokaryotes (acridine orange direct counts), which reflects the ongoing degradation of the high-molecular-weight DNA with sediment age and depth. The occurrence of eukaryotic DNA also suggests DNA preservation in the deeply buried sediments.     

The paleoclimate of the Eastern Mediterranean during the transition from early to mid Pleistocene (900 to 700 ka) based on marine and non-marine records: An integrated overview

Climate change is frequently considered an important driver of hominin evolution and dispersal patterns. The role of climate change in the last phase (900-700 ka) of the Middle Pleistocene Transition (MPT) in the Levant and northeast Africa was examined, using marine and non-marine records. During the MPT the global climate system shifted from a linear 41 k.yr. into a highly non-linear 100 k.yr. system, considerably changing its global modulation. Northeast Africa aridity further intensified around 950 ka, as indicated by a sharp increase in dust flux, and a jump to overall higher levels thereafter, coinciding with a lack of sapropels in the deep eastern Mediterranean (930-690 ka). The increased dust flux centering at ∼800 ka corresponds to the minima in 400 k.yr. eccentricity, a minima in 65 °N solar forcing and in the weakest African monsoon precession periodicity. This resulted in expansion of hyper-arid conditions across North Africa, the lowest lake levels in eastern Africa and the lowest rainfall in the Nile River headwaters. In the eastern Mediterranean an increasing continental signature is seen in glacial stages 22 (∼880 ka) and 20 (∼800 ka). Lower arboreal pollen values also indicate arid conditions during these glacial stages. The southern and eastern parts of the Negev Desert, unlike its northern part, were hyper-arid during the MPT, making them highly unsustainable. The fluctuations in the stands of Lake Amora follow global climate variability but were more moderate than those of its last glacial Lake Lisan successor. In the northern Jordan-Valley Hula Lake, frequent fluctuations in lake level coincide with both global climate changes and minor changes in water salinity varying from fresh to oligohaline. It appears therefore that the most pronounced and widespread deterioration in climate occurred in northeast Africa from 900 to 700 ka, whereas in the Levant the corresponding climatic changes were more moderate.     

The palaeoenvironments of coastal lagoons in the southern Baltic Sea,II. δ13C and δ15N ratios of organic matter — sources and sediments

Organic carbon and nitrogen isotope values (δ¹³C, δ¹⁵N) and C/N ratios of six sediment cores from six coastal lagoons (including the Oder Estuary) were measured to chart the coastal development and to reconstruct the local palaeoenvironments of the southern Baltic Sea region during the Holocene. In addition, δ¹³C, δ¹⁵N and C/N values of major organic matter sources in the coastal lagoons and their drainage areas are investigated to determine the origin of organic matter (i.e. terrigenous or marine) in the sediments: plankton, aquatic macrophytes, typical C3 shore plants and peat. The δ¹³C, δ¹⁵N and C/N values of the samples collected show the clearly identifiable stages in the development of the water bodies: post-glacial lake stages with sandy sedimentation, lacustrine phases with high autochthonous productivity, terrestrial stages with peat formation, sedimentation as a result of marine transgression, and brackish sedimentation after the formation of sand spits and barrier islands. These stages are the results of sea level changes in the region. The values allow derivation of differences in the palaeoenvironments of the lagoons in the study area. A distinct terrestrial input is evident in the sediments of the lagoonal Oder Estuary, which can be attributed to the direct inflow of the Oder River into the lagoon. The isotope and C/N values also suggest a contribution of C4 plant detritus for the water bodies in the northeastern part of the study area (Barther Bodden, Grabow). The burial of autochthonous organic matter (i.e. plankton, aquatic macrophytes) in the sediment could be derived for all lagoons in this investigation.     

Stratigraphy and tephra of the Kibish Formation, southwestern Ethiopia

The Kibish Formation in southwestern Ethiopia, with an aggregate thickness of ~105m, consists of lacustrine, marginal lacustrine, and deltaic deposits. It is divided into four members numbered I to IV on the basis of erosion surfaces (disconformities) between the strata of each member. It overlies the Mursi and Nkalabong formations, the latter of which is here shown to correlate with the Shungura Formation. Tephra layers in each member allow for secure correlation between geographically separated sections on the basis of the composition of their volcanic glass. Members I, III, and IV of the Kibish Formation appear to have been deposited at the same times as sapropels S7 (197ka), S4 (104ka), and S1 (8ka) in the eastern Mediterranean Sea, respectively. We correlate the KHS Tuff of the Kibish Formation with a >154-kyr-old unnamed tuff in the Konso Formation. Tephra in Member IV may derive from Mount Wenchi, a volcano situated on the divide between the Omo and Blue Nile drainage basins. Thin-bedded sedimentary layers probably represent annual deposition reflecting rapid sedimentation (~30m/kyr) of parts of the formation. This conclusion is supported by variation in paleomagnetic inclination through a sequence of these layers at KHS. Two fossils of early Homo sapiens (Omo I and Omo II) derive from Member I. Their stratigraphic placement is confirmed by analysis of the KHS Tuff in the lower part of Member II at both fossil sites. The KHS Tuff lies above a disconformity, which itself lies above the fossils at both sites. (40)Ar/(39)Ar dates provide an estimated age of ~195kyr for these fossils. Omo III, a third fossil H. sapiens, probably also derives from Member I of the Kibish Formation and is of similar age. Hominin fossils from AHS, a new site, also derive from Member I. Hominin fossils from CHS can only be placed between 104ka and 10ka, the H. sapiens specimen from JHS is most likely 9-13kyr in age, and a partial skeleton of H. sapiens from Pelvic Corner is most likely ~6.6kyr in age.     

Sources for sedimentary bacteriohopanepolyols as revealed by 16S rDNA stratigraphy

Bacteriohopanoids are widespread lipid biomarkers in the sedimentary record. Many aerobic and anaerobic bacteria are potential sources of these lipids which sometimes complicates the use of these biomarkers as proxies for ecological and environmental changes. Therefore, we applied preserved 16S ribosomal RNA genes to identify likely Holocene biological sources of bacteriohopanepolyols (BHPs) in the sulfidic sediments of the permanently stratified postglacial Ace Lake, Antarctica. A suite of intact BHPs were identified, which revealed a variety of structural forms whose composition differed through the sediment core reflecting changes in bacterial populations induced by large changes in lake salinity. Stable isotopic compositions of the hopanols formed from periodic acid-cleaved BHPs, showed that some were substantially depleted in ¹³C, indicative of their methanotrophic origin. Using sensitive molecular tools, we found that Type I and II methanotrophic bacteria (respectively Methylomonas and Methylocystis ) were unique to the oldest lacustrine sediments (> 9400 years BP), but quantification of fossil DNA revealed that the Type I methanotrophs, including methanotrophs related to methanotrophic gill symbionts of deep-sea cold-seep mussels, were the main precursors of the 35-amino BHPs (i.e. aminopentol, -tetrol and -triols). After isolation of the lake ∼3000 years ago, one Type I methanotroph of the 'methanotrophic gill symbionts cluster' remained the most obvious source of aminotetrol and -triol. We, furthermore, identified a Synechococcus phylotype related to pelagic freshwater strains in the oldest lacustrine sediments as a putative source of 2-methylbacteriohopanetetrol (2-Me BHT). This combined application of advanced geochemical and paleogenomical tools further refined our knowledge about Holocene biogeochemical processes in Ace Lake.     

Land Snails as a Diet Diversification Proxy during the Early Upper Palaeolithic in Europe

Despite the ubiquity of terrestrial gastropods in the Late Pleistocene and Holocene archaeological record, it is still unknown when and how this type of invertebrate resource was incorporated into human diets. In this paper, we report the oldest evidence of land snail exploitation as a food resource in Europe dated to 31.3-26.9 ka yr cal BP from the recently discovered site of Cova de la Barriada (eastern Iberian Peninsula). Mono-specific accumulations of large Iberus alonensis land snails (Ferussac 1821) were found in three different archaeological levels in association with combustion structures, along with lithic and faunal assemblages. Using a new analytical protocol based on taphonomic, microX-Ray Diffractometer (DXR) and biometric analyses, we investigated the patterns of selection, consumption and accumulation of land snails at the site. The results display a strong mono-specific gathering of adult individuals, most of them older than 55 weeks, which were roasted in ambers of pine and juniper under 375°C. This case study uncovers new patterns of invertebrate exploitation during the Gravettian in southwestern Europe without known precedents in the Middle Palaeolithic nor the Aurignacian. In the Mediterranean context, such an early occurrence contrasts with the neighbouring areas of Morocco, France, Italy and the Balkans, where the systematic nutritional use of land snails appears approximately 10,000 years later during the Iberomaurisian and the Late Epigravettian. The appearance of this new subsistence activity in the eastern and southern regions of Spain was coeval to other demographically driven transformations in the archaeological record, suggesting different chronological patterns of resource intensification and diet broadening along the Upper Palaeolithic in the Mediterranean basin.     

The mid‐Holocene vegetation of the Mediterranean region and southern Europe,and comparison with the present day

Aim To contribute to the intense debate surrounding the relative influence of climate and humans on Mediterranean‐region land cover over the past 6000 years, we assess the Holocene biogeography and vegetation history of southern Europe by means of an extensive pollen record dataset. Location The Mediterranean biogeographical zone and neighbouring parts of Iberia, the Alps and Anatolia, between 30° N, 48° N, 10° W and 45° E. Methods We compiled a southern European pollen record dataset using available pollen databases (124 sites) and other sources (74 sites), with improved spatial coverage and dating control compared with earlier studies. We used only those sites that had pollen data for both 0 ka and 6 ka. We reconstructed mid‐Holocene and present‐day biomes, arboreal pollen percentages and distribution and relative abundance of 11 key woody taxa, with anomaly maps. Results Northern temperate forest biomes extended further south at the mid‐Holocene than at present, but not as far as earlier studies suggested. Sclerophyllous vegetation occurred along the Mediterranean coast throughout the region at 6 ka. Arboreal pollen percentages were up to 50% higher than at present. At 6 ka, Olea, Fagus and Juniperus had smaller distributions and/or abundances; Abies, Cedrus and both deciduous and evergreen Quercus had larger distributions and/or abundances; Phillyrea, Pistacia and Cistus showed minimal difference; and Pinus showed a cosmopolitan distribution with variable abundance. Main conclusions Temporal difference analysis is more meaningful when only sites containing samples for all time slices are analysed. During the mid‐Holocene, southern Europe was more heavily forested with temperate vegetation than it is at present, but drought‐tolerant xeric vegetation was still widespread along the southern margins of the region. Although human land use may have caused the degradation of land between the mid‐Holocene and the present, the mere presence of xeric vegetation in the Mediterranean region does not require human impact. This challenges the commonly held belief that modern Mediterranean vegetation represents a ‘degraded’ state.     

Bacterial and Archaeal 16S rRNA Genes in Late Pleistocene to Holocene Muddy Sediments from the Kanto Plain of Japan

Microbial communities in ancient marine sediments composed of clay and silt obtained from the terrestrial subsurface were phylogenetically analyzed based on their 16S rRNA gene sequences. Chloroflexi and Miscellaneous Crenarchaeotic Group were predominant in bacterial and archaeal clone libraries, respectively. Of 44 operational taxonomic units (OTUs) that had close relatives in the database, 30 were close to sequences obtained from marine environments. Some sequences belonged to the candidate groups JS1, ANME-I, and Marine Benthic Group-C, which are typically found in marine sediments. Low chloride concentrations in the sediments suggest that these marine-affiliated sequences may not reflect currently active microbial communities. Our results indicate the existence of long-term preserved DNA or descendants of ancient oceanic microbial components in subsurface muddy sediments in a temperate region, which may reflect indigenous population of paleoenvironments.