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Human microbiome studies are increasingly incorporating macroecological approaches, such as community assembly, network analysis and functional redundancy to more fully characterize the microbiome. Such analyses have not been applied to ancient human microbiomes, preventing insights into human microbiome evolution. We address this issue by analysing published ancient microbiome datasets: coprolites from Rio Zape ( n = 7; 700 CE Mexico) and historic dental calculus ( n = 44; 1770–1855 CE, UK), as well as two novel dental calculus datasets: Maya ( n = 7; 170 BCE-885 CE, Belize) and Nuragic Sardinians ( n = 11; 1400–850 BCE, Italy). Periodontitis-associated bacteria ( Treponema denticola , Fusobacterium nucleatum and Eubacterium saphenum ) were identified as keystone taxa in the dental calculus datasets. Coprolite keystone taxa included known short-chain fatty acid producers ( Eubacterium biforme, Phascolarctobacterium succinatutens ) and potentially disease-associated bacteria ( Escherichia , Brachyspira) . Overlap in ecological profiles between ancient and modern microbiomes was indicated by similarity in functional response diversity profiles between contemporary hunter–gatherers and ancient coprolites, as well as parallels between ancient Maya, historic UK, and modern Spanish dental calculus; however, the ancient Nuragic dental calculus shows a distinct ecological structure. We detected key ecological signatures from ancient microbiome data, paving the way to expand understanding of human microbiome evolution. This article is part of the theme issue ‘Insights into health and disease from ancient biomolecules’.

The early village at Çatalhöyük (7100–6150 BC) provides important evidence for the Neolithic and Chalcolithic people of central Anatolia. This article reports on the use of lipid biomarker analysis to identify human coprolites from midden deposits, and microscopy to analyse these coprolites and soil samples from human burials. Whipworm (Trichuris trichiura) eggs are identified in two coprolites, but the pelvic soil samples are negative for parasites. Çatalhöyük is one of the earliest Eurasian sites to undergo palaeoparasitological analysis to date. The results inform how intestinal parasitic infection changed as humans modified their subsistence strategies from hunting and gathering to settled farming.

Over the past 50,000 y, biotic extinctions and declines have left a legacy of vacant niches and broken ecological interactions across global terrestrial ecosystems. Reconstructing the natural, unmodified ecosystems that preceded these events relies on high-resolution analyses of paleoecological deposits. Coprolites are a source of uniquely detailed information about trophic interactions and the behaviors, gut parasite communities, and microbiotas of prehistoric animal species. Such insights are critical for understanding the legacy effects of extinctions on ecosystems, and can help guide contemporary conservation and ecosystem restoration efforts. Here we use high-throughput sequencing (HTS) of ancient eukaryotic DNA from coprolites to reconstruct aspects of the biology and ecology of four species of extinct moa and the critically endangered kakapo parrot from New Zealand (NZ). Importantly, we provide evidence that moa and prehistoric kakapo consumed ectomycorrhizal fungi, suggesting these birds played a role in dispersing fungi that are key to NZ’s natural forest ecosystems. We also provide the first DNA-based evidence that moa frequently supplemented their broad diets with ferns and mosses. Finally, we also find parasite taxa that provide insight into moa behavior, and present data supporting the hypothesis of coextinction between moa and several parasite species. Our study demonstrates that HTS sequencing of coprolites provides a powerful tool for resolving key aspects of ancient ecosystems and may rapidly provide information not obtainable by conventional paleoecological techniques, such as fossil analyses.

In the field of archaeological parasitology, researchers have long documented the distribution of parasites in archaeological time and space through the analysis of coprolites and human remains. This area of research defined the origin and migration of parasites through presence/absence studies. By the end of the 20th century, the field of pathoecology had emerged as researchers developed an interest in the ancient ecology of parasite transmission. Supporting studies were conducted to establish the relationships between parasites and humans, including cultural, subsistence, and ecological reconstructions. Parasite prevalence data were collected to infer the impact of parasitism on human health. In the last few decades, a paleoepidemiological approach has emerged with a focus on applying statistical techniques for quantification. The application of egg per gram (EPG) quantification methods provide data about parasites’ prevalence in ancient populations and also identify the pathological potential that parasitism presented in different time periods and geographic places. Herein, we compare the methods used in several laboratories for reporting parasite prevalence and EPG quantification. We present newer quantification methods to explore patterns of parasite overdispersion among ancient people. These new methods will be able to produce more realistic measures of parasite infections among people of the past. These measures allow researchers to compare epidemiological patterns in both ancient and modern populations.

Coprolites offer rich potential for palaeodietary studies as snapshots of past dietary behaviour and environment. They require adapted laboratory methods to retrieve the DNA of the depositor, its microbiome, diet and environmental taxa. Here we compare the performance of three common ancient DNA (aDNA) extraction methods to recover metagenomes from coprolites of Darwin's ground sloth from Cueva del Milodón (Chile). The Qiagen PowerSoil Kit outperformed the other two methods in terms of DNA recovery and library complexity, but the communities inferred from the DNA extracted by the three methods were similar. We were able to recover signatures of local Patagonian flora, as well as sloth mitochondrial genomes, confirming the taxonomic identity of the coprolite depositors.

In this study, I use microbiome datasets from global soil samples and diverse hosts to learn whether soil microbial taxa are found in host microbiomes, and whether these observations fit the narrative that environmental interaction influences human microbiomes. A major motivation for conducting host-associated microbiome research is to contribute towards understanding how the environment may influence host physiology. The microbial molecular network is considered a key vector by which environmental traits may be transmitted to the host. Research on human evolution seeks evidence that can inform about the living experiences of human ancestors. This objective is substantially enhanced by recent work on ancient biomolecules from preserved microbial tissues, such as dental calculus, faecal sediments and whole coprolites. A challenge yet is to distinguish authentic biomolecules from environmental contaminants deposited contemporaneously, primarily from soil. However, we do not have sound expectations about the soil microbial elements arriving to host-associated microbiomes in a modern context. One assumption in human microbiome research is that proximity to the natural environment should affect biodiversity or impart genetic elements. I present evidence supporting the assumption that environmental soil taxa are found among host-associated gut taxa, which can recapitulate the surrounding host habitat ecotype. Soil taxa found in gut microbiomes relate to a set of universal ‘core’ taxa for all soil ecotypes, demonstrating that widespread host organisms may experience a consistent pattern of external environmental cues, perhaps critical for development. Observed differentiation of soil feature diversity, abundance and composition among human communities, great apes and invertebrate hosts also indicates that lifestyle patterns are inferable from an environmental signal that is retrievable from gut microbiome amplicon data. This article is part of the theme issue ‘Insights into health and disease from ancient biomolecules’.

Coprolites contain various kinds of ancient DNAs derived from gut micro-organisms, viruses, and foods, which can help to determine the gut environment of ancient peoples. Their genomic information should be helpful in elucidating the interaction between hosts and microbes for thousands of years, as well as characterizing the dietary behaviors of ancient people. We performed shotgun metagenomic sequencing on four coprolites excavated from the Torihama shell-mound site in the Japanese archipelago. The coprolites were found in the layers of the Early Jomon period, corresponding stratigraphically to 7000 to 5500 years ago. After shotgun sequencing, we found that a significant number of reads showed homology with known gut microbe, viruses, and food genomes typically found in the feces of modern humans. We detected reads derived from several types of phages and their host bacteria simultaneously, suggesting the coexistence of viruses and their hosts. The food genomes provide biological evidence for the dietary behavior of the Jomon people, consistent with previous archaeological findings. These results indicate that ancient genomic analysis of coprolites is useful for understanding the gut environment and lifestyle of ancient peoples.

Fungal spores that grew on the faeces of herbivores in the past can be extracted from sediments and used to identify the presence of herbivores in former ecosystems. This review: (i) examines the factors that should be considered when interpreting these fungal spores, (ii) assesses the degree to which they can be used to estimate past herbivore populations and biomass density change, and (iii) identifies gaps in our current understanding that limit, or confound, the information that can be extracted from the fungal spore record. We focus on the life cycles of coprophilous fungi and highlight the importance of understanding spore dispersal mechanisms to ensure robust palaeoecological interpretation. We then discuss how variation in methodological approaches across studies and modifications can influence comparability between studies. The key recommendations that emerge relate to: (i) improving our understanding of the relationship between spores of coprophilous fungi (SCF) and herbivores through the study of the coprophilous fungi succession; (ii) refining our understanding of how climate and environment parameters effect fungal spore abundance, with particular reference to estimating past herbivore biomass density; and (iii) enhancing sedimentary DNA (SedaDNA) analysis to identify SCF that do not allow preservation in a way that allows visual identification. To further this field of study and provide more robust insights into herbivores in the past, we suggest that additional research is required to help to reduce bias during the preparation process, that concertation metrics are used for the quantification of SCF, and that multiple cores should be taken in each site and multiproxy analysis should be utilised.

The aim of this research was to analyse the variability of parasite assemblages on a small spatial scale, by examining carnivore coprolites from the archaeological site Campo Moncada 2 (CM2), Piedra Parada area, Chubut province, Argentina, and comparing the results with those previously obtained from the archaeological site Campo Cerda 1 (CCe1), located in the same area. Six carnivore coprolites from CM2 were analysed: 4 obtained in sub-level 2a and 2 obtained in sub-level 2a/b. Two radiocarbon dates associated with the coprolites placed the samples chronologically between 780 ± 80 and 860 ± 80 years before present. The rehydrated sediments were sieved and then allowed to sediment spontaneously. The sediment was used for parasitological examination under light microscopy. Conservative estimation of total parasite richness resulted in 21 parasitic taxa. The taxa with the highest fecal prevalence (>50%) corresponded to parasites prevalent in modern carnivores (Alaria sp., Toxocara cf. canis, Toxascaris sp., Eucoleus cf. aerophila, Trichuris sp. and Ancylostomatidae gen. sp.). Assuming that the fox coprolites are contemporaneous, the total fecal parasite richness estimated for CM2 and previously for CCe1 was similar. The high total parasite richness found suggests a network of host–parasite relationships that could include regional hunter-gatherers. The results obtained in carnivore coprolites allow us to infer a very diverse biological community in Piedra Parada area, so the regional caves and rockshelters could have a proportional epidemiological importance as parasite exchange nodes.