Abstract The environmental changes (vegetation history, human impact and land use, influence of the Black Sea) in the area of Lake Durankulak, northeastern Bulgaria, were reconstructed and synthesized for the last ca. 8000 years. The palaeoecological information derived from various proxies (pollen, plant macrofossils, molluscs, sediments) was compared on a regional scale with the evidence from the nearby coastal lakes Shabla–Ezeretz and Bolata. The Early Holocene xerothermic steppe vegetation, dominated by Chenopodiaceae, Artemisia and Poaceae species, and accompanied by stands of trees in moister habitats, was transformed after 6000 cal. BP into a forest-steppe, comprising oak woods with Carpinus betulus, Ulmus, Tilia, Acer. This vegetation pattern has been periodically modified, depleted and replaced by arable land or xerothermic herbaceous communities enriched with anthropophytes and ruderals, particularly after the intensification of human activities since 3300 cal. BP. The archaeobotanical evidence from the region has provided valuable information about the occupation phases and subsistence strategy of the local people since the Late Neolithic (5300 cal. BC/7250 cal. BP). Periods with cultivation of cereals (Triticum, Hordeum) and/or stock-breeding activity were interrupted by abandonment of the settlements and the arable land due to unfavourable environmental changes. The periodical connection/isolation of Lake Durankulak with the Black Sea and the periods of marine influence were recorded by changes in the composition of the fossil molluscan fauna and the lithology of the sediments, and chronologically confirmed by radiocarbon dates. The development of the coastal lakes throughout the largest part of the Holocene has been also considerably influenced by the fluctuations of the Black Sea level.
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Dental calculus metagenomics has emerged as a valuable tool for studying the oral microbiomes of humans and a few select mammals. With increasing interest in wild animal microbiomes, it is important to understand how widely this material can be used across the mammalian tree of life, refine the related protocols and understand the expected outcomes and potential challenges of dental calculus sample processing. In this study, we significantly expand the breadth of studied host species, analysing laboratory and bioinformatics metadata of dental calculus samples from 32 ecologically and phylogenetically diverse mammals. Although we confirm the presence of an oral microbiome signature in the metagenomes of all studied mammals, the fraction recognised as oral varies between host species, possibly because of both biological differences and methodological biases. The overall success rate of dental calculus processing, from extractions to sequencing, was 74\%. Although input sample weight was positively associated with the number of produced library molecules, we identify a negative impact of enzymatic inhibition on the library preparation protocol. The inhibition was most prevalent in herbivores and frugivores and is likely diet-derived. In contrast, hosts with an animalivore diet posed fewer challenges during laboratory processing and yielded more DNA relative to sample weight. Our results translate into recommendations for future studies of dental calculus metagenomics from a variety of host species, identifying required sample amounts, and emphasising the utility of dental calculus in exploring the oral microbiome in relation to broader ecological and evolutionary questions.
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RBINS Staff Publications 2025
