Crop manuring and intensive land management by Europe's first farmers.

The spread of farming from western Asia to Europe had profound long-term social and ecological impacts, but identification of the specific nature of Neolithic land management practices and the dietary contribution of early crops has been problematic. Here, we present previously undescribed stable isotope determinations of charred cereals and pulses from 13 Neolithic sites across Europe (dating ca. 5900-2400 cal B.C.), which show that early farmers used livestock manure and water management to enhance crop yields. Intensive manuring inextricably linked plant cultivation and animal herding and contributed to the remarkable resilience of these combined practices across diverse climatic zones. Critically, our findings suggest that commonly applied paleodietary interpretations of human and herbivore δ(15)N values have systematically underestimated the contribution of crop-derived protein to early farmer diets.

Agropastoral and dietary practices of the northern Levant facing Late Holocene climate and environmental change: Isotopic analysis of plants, animals and humans from Bronze to Iron Age Tell Tweini.

One of the largest isotopic datasets of the ancient Eastern Mediterranean region is evaluated, based on plants (n = 410), animals (n = 210) and humans (n = 16) from Tell Tweini (Syria). Diachronic analysis of plant and faunal specimens from four main periods of occupation: Early Bronze Age (2600-2000 BC), Middle Bronze Age (2000-1600 BC), Late Bronze Age (1600-1200 BC) and Iron Age (1200-333 BC) were investigated. Mean Δ13C results from seven plant species reveal emmer and free threshing wheat, olives, bitter vetch, rye grass and barley were adequately or well-watered during all periods of occupation. The grape Δ13C results suggest excellent growing conditions and particular care for its cultivation. The δ15N results indicate that especially the emmer and free threshing wheats received some manure inputs throughout the occupation sequence, while these were likely further increased during the Iron Age, encompassing also the olive groves and grape vineyards. Generally, domestic animals (cattle, sheep, goats) had C3 terrestrial diets and were kept together in similar environments. However, some animals consumed significant amounts of marine or C4 plants, possibly from disturbed habitats due to land use pressure or salt tolerant grasses and shrubs from wetland environments, which were recorded in the direct vicinity of the site. Middle Bronze Age humans consumed a C3 terrestrial diet with no measurable input from C4, freshwater or marine protein sources. Interestingly, the human diet was relatively low in animal protein and appears comparable to what is considered today a typical Mediterranean diet consisting of bread (wheat/barley), olives, grapes, pulses, dairy products and small amounts of meat. The combined isotopic analysis of plants, animals and humans from Tell Tweini represents unbroken links in the food chain which create unparalleled opportunities to enhance our current understanding of environmental conditions, climate change and lifeways in past populations from the Eastern Mediterranean.

Melatonin Receptor 1B and Corticosteroid Receptor Polymorphisms in Infertile Women with Implantation Failure and Miscarriages.

BACKGROUND: The development of assisted reproductive techniques has significantly improved fertility chances in many women, but recurrent implantation failure (RIF) and miscarriages (RM) might preclude successful pregnancy. Alterations in the intrinsic secretory patterns of melatonin and cortisol influence reproduction in humans, and imperfection of receptor - dependent signaling may additionally compromise the hormonal effects. Therefore, the present study aims to investigate the influence of certain melatonin and cortisol receptor polymorphisms in infertile women. METHODS: A total of 111 female infertile patients suffering from implantation failure and/or miscarriages were genotyped for MTNR1B rs1562444, MTNR1Brs10830962, GCCR rs41423247, and GCCR ER22/23EK variants. Additionally, 106 female volunteers were genotyped for the same polymorphisms. RESULTS: The allele and genotype distribution of the investigated polymorphisms did not differ between infertile women and the control group. Significantly more women with history of RIF have MTNR1B rs1562444 G-allele-containing genotypes in comparison to AA carriers (19.3% vs. 3.6%, p = 0.004). The minor allele of the ER22/23EK variant was more frequent in infertile patients with three or more unsuccessful implantation attempts than in other women (12.5% vs. 2.4%, p = 0.025). CONCLUSIONS: Melatonin receptor 1B polymorphisms might affect embryo implantation and early pregnancy loss, while their influence on late pregnancy complications needs further evaluation. The possible association between the cortisol receptor ER22/23EK variant and recurrent implantation failure might help to differentiate women who could benefit from corticosteroid treatment.

Isotopic reconstruction of human diet and animal husbandry practices during the Classical-Hellenistic, imperial, and Byzantine periods at Sagalassos, Turkey.

An isotopic reconstruction of human dietary patterns and livestock management practices (herding, grazing, foddering, etc.) is presented here from the sites of Düzen Tepe and Sagalassos in southwestern Turkey. Carbon and nitrogen stable isotope ratios were determined from bone collagen extracted from humans (n = 49) and animals (n = 454) from five distinct time periods: Classical-Hellenistic (400-200 BC), Early to Middle Imperial (25 BC-300 AD), Late Imperial (300-450 AD), Early Byzantine (450-600 AD), and Middle Byzantine (800-1200 AD). The humans had protein sources that were based on C(3) plants and terrestrial animals. During the Classical-Hellenistic period, all of the domestic animals had δ(13) C and δ(15) N signatures that clustered together; evidence that the animals were herded in the same area or kept in enclosures and fed on similar foods. The diachronic analysis of the isotopic trends in the dogs, cattle, pigs, sheep, and goats highlighted subtle but distinct variations in these animals. The δ(13) C values of the dogs and cattle increased (reflecting C(4) plant consumption) during the Imperial and Byzantine periods, but the pigs and the goats displayed little change and a constant C(3) plant-based diet. The sheep had a variable δ(13) C pattern reflecting periods of greater and lesser consumption of C(4) plants in the diet. In addition, the δ(15) N values of the dogs, pigs, cattle, and sheep increase substantially from the Classical-Hellenistic to the Imperial periods reflecting a possible increase in protein consumption, but the goats showed a decrease. Finally, these isotopic results are discussed in the context of zooarcheological, archeobotanical, and trace element evidence.

A new method based on surface-sample pollen data for reconstructing palaeovegetation patterns.

Aim: Biomisation has been the most widely used technique to reconstruct past regional vegetation patterns because it does not require an extensive modern pollen dataset. However, it has well-known limitations including its dependence on expert judgement for the assignment of pollen taxa to plant functional types (PFTs) and PFTs to biomes. Here we present a new method that combines the strengths of biomisation with those of the alternative dissimilarity-based techniques. Location: The Eastern Mediterranean-Black Sea Caspian Corridor (EMBSeCBIO). Taxon: Plants. Methods: Modern pollen samples, assigned to biomes based on potential natural vegetation data, are used to characterize the within-biome means and standard deviations of the abundances of each taxon. These values are used to calculate a dissimilarity index between any pollen sample and every biome, and thus assign the sample to the most likely biome. We calculate a threshold value for each modern biome; fossil samples with scores below the threshold for all modern biomes are thus identified as non-analogue vegetation. We applied the new method to the EMBSeCBIO region to compare its performance with existing reconstructions. Results: The method captured changes in the importance of individual taxa along environmental gradients. The balanced accuracy obtained for the EMBSeCBIO region using the new method was better than obtained using biomisation (77% vs. 65%). When the method was applied to high-resolution fossil records, 70% of the entities showed more temporally stable biome assignments than obtained using biomisation. The technique also identified likely non-analogue assemblages in a synthetic modern dataset and in fossil records. Main conclusions: The new method yields more accurate and stable reconstructions of vegetation than biomisation. It requires an extensive modern pollen dataset, but is conceptually simple, and avoids subjective choices about taxon allocations to PFTs and PFTs to biomes.

Mashes to Mashes, Crust to Crust. Presenting a novel microstructural marker for malting in the archaeological record.

The detection of direct archaeological remains of alcoholic beverages and their production is still a challenge to archaeological science, as most of the markers known up to now are either not durable or diagnostic enough to be used as secure proof. The current study addresses this question by experimental work reproducing the malting processes and subsequent charring of the resulting products under laboratory conditions in order to simulate their preservation (by charring) in archaeological contexts and to explore the preservation of microstructural alterations of the cereal grains. The experimentally germinated and charred grains showed clearly degraded (thinned) aleurone cell walls. The histological alterations of the cereal grains were observed and quantified using reflected light and scanning electron microscopy and supported using morphometric and statistical analyses. In order to verify the experimental observations of histological alterations, amorphous charred objects (ACO) containing cereal remains originating from five archaeological sites dating to the 4th millennium BCE were considered: two sites were archaeologically recognisable brewing installations from Predynastic Egypt, while the three broadly contemporary central European lakeshore settlements lack specific contexts for their cereal-based food remains. The aleurone cell wall thinning known from food technological research and observed in our own experimental material was indeed also recorded in the archaeological finds. The Egyptian materials derive from beer production with certainty, supported by ample contextual and artefactual data. The Neolithic lakeshore settlement finds currently represent the oldest traces of malting in central Europe, while a bowl-shaped bread-like object from Hornstaad-Hörnle possibly even points towards early beer production in central Europe. One major further implication of our study is that the cell wall breakdown in the grain's aleurone layer can be used as a general marker for malting processes with relevance to a wide range of charred archaeological finds of cereal products.

New AMS 14C dates track the arrival and spread of broomcorn millet cultivation and agricultural change in prehistoric Europe.

Broomcorn millet (Panicum miliaceum L.) is not one of the founder crops domesticated in Southwest Asia in the early Holocene, but was domesticated in northeast China by 6000 BC. In Europe, millet was reported in Early Neolithic contexts formed by 6000 BC, but recent radiocarbon dating of a dozen 'early' grains cast doubt on these claims. Archaeobotanical evidence reveals that millet was common in Europe from the 2nd millennium BC, when major societal and economic transformations took place in the Bronze Age. We conducted an extensive programme of AMS-dating of charred broomcorn millet grains from 75 prehistoric sites in Europe. Our Bayesian model reveals that millet cultivation began in Europe at the earliest during the sixteenth century BC, and spread rapidly during the fifteenth/fourteenth centuries BC. Broomcorn millet succeeds in exceptionally wide range of growing conditions and completes its lifecycle in less than three summer months. Offering an additional harvest and thus surplus food/fodder, it likely was a transformative innovation in European prehistoric agriculture previously based mainly on (winter) cropping of wheat and barley. We provide a new, high-resolution chronological framework for this key agricultural development that likely contributed to far-reaching changes in lifestyle in late 2nd millennium BC Europe.

The Hoard of the Rings. "Odd" annular bread-like objects as a case study for cereal-product diversity at the Late Bronze Age hillfort site of Stillfried (Lower Austria).

Cereals, in addition to being a major ingredient in daily meals, also play a role in the preparation of foodstuffs for ritual purposes. This paper deals with finds that may correspond to such ritual preparations retrieved from the hillfort site of Stillfried an der March. The site, spreading across an area of ca. 23 ha, held a very important position among settlements of Late Urnfield period (particularly during the 10th- 9th c. BCE), acting as a central place where large scale storage of grain as well as textile and metal production took place under the control of local elites. Three incomplete ring-shaped charred organic objects, found together with 14 rings and ring fragments made of clay were discovered in a secondary filled silo pit, excavated among a total of about 100 pits of this kind at the site. The overall good state of preservation of the organic ring fragments suggests that they were deposited intact on the bottom of the pit and covered well so that no re-deposition or damage occurred. This could be indicate their intentional placement in this position. Light and scanning electron microscopy revealed that the charred organic rings are cereal products containing hulled barley and a wheat species. Indications that the objects were shaped from a wet cereal mixture and had been subsequently dried without baking are discussed, as well as the possible significance of the find assemblage. The annular objects are put in context with the contemporary cereal spectrum as well as other cereal preparations from Stillfried, outlining their different chaînes opératoires for handling cereal food.

Brassicaceae seed oil identified as illuminant in Nilotic shells from a first millennium AD Coptic church in Bawit, Egypt.

Burned greasy deposits were found inside shells of the large Nile bivalve Chambardia rubens, excavated in an eight- to tenth- century AD church of the Coptic monastery of Bawit, Egypt, and supposedly used as oil lamps. The residues were subjected to a combination of chromatographic residue analysis techniques. The rather high concentrations of unsaturated fatty acids, as analysed by gas chromatography (GC) in the methylated extract, suggest the presence of a vegetal oil. Analysis of the stable carbon isotopes (delta 13C values) of the methyl esters also favoured plants over animals as the lipid source. In the search for biomarkers by GC coupled to mass spectrometry on a silylated extract, a range of diacids together with high concentrations of 13,14-dihydroxydocosanoate and 11,12-dihydroxyeicosanoate were found. These compounds are oxidation products of erucic acid and gondoic acid, which are abundantly present in seeds of Brassicaceae plants. Liquid chromatography coupled to mass spectrometry analysis showed low concentrations of unaltered triglycerides, but revealed sizeable amounts of triglycerides with at least one dihydroxylated acyl chain. The unusual preservation of dihydroxylated triglycerides and alpha,omega-dicarboxylic acids can be related to the dry preservation conditions. Analysis of the stereoisomers of the dihydroxylated fatty acids allows one to determine whether oxidation took place during burning of the fuel or afterwards. The results prove that the oil of rapeseed (Brassica napus L.) or radish (Raphanus sativus L.) was used as illuminant in early Islamic Egypt, and that not only ceramic lamps but also mollusk shells were used as fuel containers.

Correction: Pioneer farming in southeast Europe during the early sixth millennium BC: Climate-related adaptations in the exploitation of plants and animals.

[This corrects the article DOI: 10.1371/journal.pone.0197225.].

Pioneer farming in southeast Europe during the early sixth millennium BC: Climate-related adaptations in the exploitation of plants and animals.

The Old World farming system arose in the semi-arid Mediterranean environments of southwest Asia. Pioneer farmers settling the interior of the Balkans by the early sixth millennium BC were among the first to introduce southwest Asian-style cultivation and herding into areas with increasingly continental temperate conditions. Previous research has shown that the bioarchaeological assemblages from early farming sites in southeast Europe vary in their proportions of plant and animal taxa, but the relationship between taxonomic variation and climate has remained poorly understood. To uncover associations between multiple species and environmental factors simultaneously, we explored a dataset including altitude, five bioclimatic and 30 bioarchaeological variables (plant and animal taxa) for 57 of the earliest farming sites in southeast Europe using Canonical Correspondence Analysis (CCA). An extension of correspondence analysis, CCA is widely used in applied ecology to answer similar questions of species-environment relationships, but has not been previously applied in prehistoric archaeology to explore taxonomic and climatic variables in conjunction. The analyses reveal that the changes in plant and animal exploitation which occurred with the northward dispersal of farmers, crops and livestock correlate with south-north climate gradients, and emphasize the importance of adaptations in the animal domain for the initial establishment of farming beyond the Mediterranean areas.

State of the (t)art. Analytical approaches in the investigation of components and production traits of archaeological bread-like objects, applied to two finds from the Neolithic lakeshore settlement Parkhaus Opéra (Zürich, Switzerland).

The site of Parkhaus Opéra is located on the north-eastern shore of Lake Zürich (Switzerland) and was documented during a rescue excavation in 2010 and 2011 by the Office for Urbanism, City of Zürich. Two charred bread-like objects were found in late Neolithic Layer 13 of the pile-dwelling, and are investigated using a novel set of analyses for cereal-based foodstuffs. Tissue remains of barley and wheat were identified, as well as a schizocarp of celery (cf. Apium graveolens), providing the first evidence for the use of bread condiments in the Neolithic. Cereal particle sizes were recorded and used to draw conclusions regarding milling and sieving of the raw material. Gas bubbles in the charred objects were measured in order to evaluate possible leavening of the dough. The outcomes of this research significantly advance the understanding of the production traits of cereal-based food during the Neolithic. The analytical techniques proposed by this study open up new possibilities for systematic and consistent investigations of cereal-based archaeological foodstuffs.