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Reconstruction of ancient epigenomes by DNA methylation (DNAm) can shed light into the composition of cell types, disease states, and age at death. However, such analysis is hampered by impaired DNA quality and little is known how decomposition affects DNAm. In this study, we determined if EPIC Illumina BeadChip technology is applicable for specimens from mummies of the eighteenth century CE. Overall, the signal intensity on the microarray was extremely low, but for one of two samples we were able to detect characteristic DNAm signals in a subset of CG dinucleotides (CpGs), which were selected with a stringent processing pipeline. Using only these CpGs we could train epigenetic signatures with reference DNAm profiles of multiple tissues and our predictions matched the fact that the specimen was lung tissue from a 28-year-old woman. Thus, we provide proof of principle that Illumina BeadChips are applicable for DNAm profiling in ancient samples.

Our previous research on coprolite specimens from the mummies of Joseon Dynasty (1392-1910 CE) has revealed various species of parasite eggs. Herein, we added 2 new helminthic cases of human remains from Joseon-period graves in the Republic of Korea (Korea). The organic materials precipitated on the hip bones of 2 half-mummied cases (Goryeong and Gwangmyeong cases) were collected, rehydrated, and examined by a microscope. In the sample from Goryeong-gun (gun=County), ova of Trichuris trichiura, Clonorchis sinensis, and Metagonimus spp. were detected, and eggs of T. trichiura and A. lumbricoides were found from the sample of Gwangmyeong-si (si=City). By adding this outcome to the existing data pool, we confirm our previous estimates of Joseon-period parasite infection rates. The overall rates of A. lumbricoides, T. trichiura, and C. sinensis decreased dramatically from Joseon to the modern period. In Goryeong mummy specimen, we also found Metagonimus spp. eggs that has rarely been detected in archaeological samples so far.

The molecular Egyptology field started in the mid-eighties with the first publication on the ancient DNA (aDNA) analysis of an Egyptian mummy. Egypt has been a major interest for historians, archeologists, laymen as well as scientists. The aDNA research on Egyptian biological remains has been fueled by their abundance and relatively well-preserved states through artificial mummification and by the advanced analytical techniques. Early doubts of aDNA integrity within the Egyptian mummies and data authenticity were later abated with studies proving successfully authenticated aDNA retrieval. The current review tries to recapitulate the published studies presenting paleogenomic evidence of disease diagnosis and kinship establishment for the Egyptian human remains. Regarding disease diagnosis, the prevailing literature was on paleogenomic evidence of infectious diseases in the human remains. A series of reports presented evidence for the presence of tuberculosis and/or malaria. In addition, there were solitary reports of the presence of leprosy, diphtheria, bacteremia, toxoplasmosis, schistosomiasis and leishmaniasis. On the contrary, paleogenomic evidence of the presence of rare diseases was quite scarce and mentioned only in two articles. On the other hand, kinship analysis of Egyptian human remains, including that of Tutankhamen, was done using both mitochondrial DNA sequences and nuclear DNA markers, to establish family relationships in four studies. It is clear that the field of molecular Egyptology is still a largely unexplored territory. Nevertheless, the paleogenomic investigation of Egyptian remains could make significant contributions to biomedical sciences (e.g. elucidation of coevolution of human host–microbe interrelationship) as well as to evidence-based archeology.

In an examination of three unwrapped mummified children from the Graeco-Roman Period of ancient Egypt there was an unexpected finding of fair hair. In the majority of unwrapped ancient Egyptian mummified bodies the hair was not fair but rather dark brown or black. To determine if exposure to natron during the mummification process was responsible for the fair hair color an experiment was carried out to partially replicate the environment in which bodies were desiccated. Fourteen samples of modern hair from various age groups, sex and ethnicity were subjected to synthetic natron for a period of 40 days to replicate the time taken to mummify a body. Macroscopic and microscopic examinations of samples were employed to ascertain any significant changes in hair color after treatment. Ancient wigs were studied for evidence of post mortem changes to hair color since construction over 2,000 years ago. Results of the study showed no significant lightening of hair color and in several samples the hair significantly darkened as the result of exposure to the natron. There was not any evidence that hair lightened as the result of natural post mortem changes and this was confirmed by the study of the natural hair wigs that had not changed color post mortem. This study concluded that the fair hair observed in the three child mummies was not the result of exposure to natron or post mortem changes but rather it was probably due to ancestry because of the presence of diverse genomes that were introduced into ancient Egypt during the Greco-Roman Period.

The identity of the mummified Lady from the Barfusser Church in Basel, Switzerland has been unsolved for decades, despite the prominent location of the burial place in front of the choir screen. A recent multidisciplinary research approach came up with a possible candidate, Anna Catharina Bischoff who died in Basel in 1787 with an age of 69 years (1719–1787). To verify the identity of the mummy, genealogists of the Citizen Science Basel discovered three living individuals of the maternal lineage of two different family branches, separated from Anna Catharina Bischoff by up to 22 generations. In this study we compare the ancient mitochondrial DNA of the mummy recovered from a premolar to the mitochondrial DNA of these three candidates. Initially the mitochondrial hypervariable regions I and II of the living individuals were screened using the Sanger sequencing method. This was followed by a mitochondrial capture approach and next generation sequencing to enrich for the whole mitochondrial genome of the mummy and one living person. A full mitochondrial genome has been recovered of both individuals sharing an identical haplotype. The sequence was assigned to the mitochondrial haplogroup U5a1+!16192 including two private mutations 10006G and 16293C. Only by using an interdisciplinary approach combining ancient DNA analysis and genealogy a maternal lineage of a non-noble family spanning 22 generations could be confirmed.

Artificial mummification has been used since antiquity and is best known from ancient Egypt. Despite ancient Egyptian mummies being studied for several decades, the mummification techniques of that time are not well understood. Modern mummification experiments involving animal and human tissues have contributed additional insights relevant to a broad field of research. In the current study, we present follow‐up results of an experiment on artificial mummification, which began in 2009. A human leg was artificially mummified and monitored during almost a year with histological, molecular, and radiological techniques. Since then, it has remained in a dry, natron salt blend for nine years. The current analyses show further progression of dehydration and tissue alterations, as well as DNA degradation, suggesting an ongoing process. Our results add new insights into the mechanisms of tissue mummification. Taking into account that the process is still ongoing, further research is required, including a re‐evaluation of the human leg in the future.

This paper integrates our knowledge from traditional Chinese medical texts and archeological findings to discuss parasitic loads in early China. Many studies have documented that several different species of eukaryotic endoparasites were present in early human populations throughout China. Nevertheless, comprehensive paleoparasitological records from China are patchy, largely due to taphonomic and environmental factors. An examination of early Chinese medical texts allows us to fill in some of the gaps and counteract apparent biases in the current archeoparasitological records. By integrating the findings of paleoparasitology with historic textual sources, we show that parasites have been affecting the lives of humans in China since ancient times. We discuss the presence and prevalence of three groups of parasites in ancient China: roundworm (Ascaris lumbricoides), Asian schistosoma (Schistosoma japonicum), and tapeworm (Taenia sp.). We also examine possible factors that favored the spread of these endoparasites among early humans. Therefore, this paper not only aims to reveal how humans have been affected by endoparasites, but also addresses how early medical knowledge developed to cope with the parasitic diseases. Nanyang Technological University Published version This research was fully supported by NAP Start-Up Grant from Nanyang Technological University. We thank our assistant, Zhu Chuan, from the School of Art, Design and Media who provided expertise on illustrating images that greatly assisted the research. We also truly thank the editors from International Journal of Paleopathology for their great works and assistance.

The Skiles Mummy (SMM), a naturally mummified adult male from the late archaic period of Lower Pecos Canyonlands of South Texas, represents a unique case of care. SMM is an exceptional mummy within this region due to both the retention of a full head of hair, and having a diagnosed case of megacolon, a complication commonly associated with Chagas disease caused by Trypanosoma cruzi. Stable isotopic analysis of his hair is consistent with a diet incorporating of C4/CAM plants with some C3 plants, freshwater resources, and higher trophic level animals. However, the segments of hair most proximal to the scalp exhibited elevated δ15N values. Data from previous research indicate starvation and malnutrition can cause δ15N values to rise. The presence of large fecal boluses in the digestive tract suggest peristalsis ceased in the last four to five months of life, and this, together with results from coprolite analysis, indicate he would not have been able to adequately absorb protein and nutrients during this time. His condition would have rendered him immobile. Following Tilley's index of care, someone would have had to bring him food resources, as well as attending to his daily needs.

International audience; Mummified remains have long attracted interest as a potential source of ancient DNA. However, mummification is a rare process that requires an anhydrous environment to rapidly dehydrate and preserve tissue before complete decomposition occurs. We present the whole-genome sequences (3.94 X) of an approximately 1600-year-old naturally mummified sheep recovered from Chehrābād, a salt mine in northwestern Iran. Comparative analyses of published ancient sequences revealed the remarkable DNA integrity of this mummy. Hallmarks of postmortem damage, fragmentation and hydrolytic deamination are substantially reduced, likely owing to the high salinity of this taphonomic environment. Metagenomic analyses reflect the profound influence of high-salt content on decomposition; its microbial profile is predominated by halophilic archaea and bacteria, possibly contributing to the remarkable preservation of the sample. Applying population genomic analyses, we find clustering of this sheep with Southwest Asian modern breeds, suggesting ancestry continuity. Genotyping of a locus influencing the woolly phenotype showed the presence of an ancestral ‘hairy’ allele, consistent with hair fibre imaging. This, along with derived alleles associated with the fat-tail phenotype, provides genetic evidence that Sasanian-period Iranians maintained specialized sheep flocks for different uses, with the ‘hairy’, ‘fat-tailed’-genotyped sheep likely kept by the rural community of Chehrābād's miners.

Mummified remains of relevant historical figures are nowadays an important source of information to retrace data concerning their private life and health, especially when historical archives are not available. Next-generation-sequencing was proved to be a valuable tool to unravel the characteristics of these individuals through their genetic heritage. Using the strictest criteria currently available for the validation of ancient DNA sequences, whole-genome and whole-exome sequencing were generated from the mummy remains of an Italian nobleman died almost 700 years ago, Cangrande della Scala. While its genome sequencing could not yield sufficient coverage for in depth investigation, exome sequencing could overcome the limitations of this approach to achieve significantly high coverage on coding regions, thus allowing to perform the first extensive exome analysis of a mummy genome. Similar to a standard “clinical exome analysis” conducted on modern DNA, an in-depth variant annotation, high-quality filtering and interpretation was performed, leading to the identification of a genotype associated with late-onset Pompe disease (glycogen storage disease type II). This genetic diagnosis was concordant with the limited clinical history available for Cangrande della Scala, who likely represents the earliest known case of this autosomal recessive metabolic disorder.

Parasitism is inherent to life and observed in all species. Extinct animals have been studied to understand what they looked like, where and how they lived, what they fed on, and the reasons they became extinct. Paleoparasitology helps to clarify these questions based on the study of the parasites and microorganisms that infected those animals, using as a source material coprolites, fossils in rock, tissue, bone, mummy, and amber, analyses of ancient DNA, immunodiagnosis, and microscopy.

Summary Paleopathology, the science that studies the diseases of the past, has always been addressed to the future in the use of new diagnostic methods. One of its relatively recent branches is paleogenetics, which is the study of genetic material from the past. Nuclear and mitochondrial DNA recovered from archaeological and paleontological specimens is called ancient DNA (aDNA), which can be extracted from a large variety of biological materials, of different origin, state of preservation and age, such as bones, teeth, coprolites, mummified tissues and hairs. There are many applications for ancient DNA research in the field of archaeology and paleopathology: population demography, genealogy, disease studies, archaeological reconstruction of plant vegetation, calibration of the molecular clock, phylogenetic relationship between different mammals and interpretation of the paleoclimate. However, the study of ancient genetic material is extremely difficult due to its poor quality and quantity, as well possible contamination with modern DNA. New advanced methods will allow extracting DNA from a greater variety of materials, and improvements in sequencing techniques will unveil data that are currently concealed. The aim of this paper is to provide initial insights into paleogenetics and ancient DNA study and to illustrate the limits, risks and potentiality of the research on the genetic material of ancient specimens, whose results have a strong impact on the present and future medicine.

Metagonimiasis is foodborne intestinal parasitism occurring by the definitive hosts' ingestion of raw or undercooked fish, mostly commonly sweetfish. Most Metagonimus infection is caused by Metagonimus yokogawai but also rarely by Metagonimus takahashii as well as Metagonimus miyatai. Despite recent molecular work on Metagonimus spp., there are still insufficient data to reveal the genetic characteristics of ancient M. yokogawai in a wide geo-historical scope. In this study, we were successful in the analysis of M. yokogawai ancient DNA (aDNA) using coprolite samples retrieved from 16th- to 17th-century Korean mummies. In BLAST and phylogenetic analyses, M. yokogawai 28S rDNA of Korean mummies were clustered along with the 28S rDNA taxa of M. takahashii and M. miyatai in GenBank. Conversely, the cytochrome c oxidase subunit I (COI) of M. yokogawai aDNA from Korean mummies was distinctly clustered apart from M. takahashii and M. miyatai sequences. This study is the first report of its kind to identify M. yokogawai aDNA retrieved from the archaeological specimens and confirms the usefulness of COI in molecular diagnosis of M. yokogawai. Considering the rarity of reports on the genetics of genus Metagonimus spp., our study will be fundamental for the future study of M. yokogawai paleogenetics.

BACKGROUND: Recent advances in sequencing have facilitated large-scale analyses of the metagenomic composition of different samples, including the environmental microbiome of air, water, and soil, as well as the microbiome of living humans and other animals. Analyses of the microbiome of ancient human samples may provide insights into human health and disease, as well as pathogen evolution, but the field is still in its very early stages and considered highly challenging. - RESULTS: The metagenomic and pathogen content of Egyptian mummified individuals from different time periods was investigated via genetic analysis of the microbial composition of various tissues. The analysis of the dental calculus’ microbiome identified Red Complex bacteria, which are correlated with periodontal diseases. From bone and soft tissue, genomes of two ancient pathogens, a 2200-year-old Mycobacterium leprae strain and a 2000-year-old human hepatitis B virus, were successfully reconstructed. - CONCLUSIONS: The results show the reliability of metagenomic studies on Egyptian mummified individuals and the potential to use them as a source for the extraction of ancient pathogen DNA. Background Results - Sample information and dating - General metagenomic assessment - Mycobacterium leprae (individual Abusir1630) - Hepatitis B virus (individual Abusir1543) - Oral microbiome assessment Discussion Conclusions Methods - Sample extraction and radiocarbon dating - Sample extraction and library preparation - Metagenomic screening - Authentication of ancient DNA - Content of endogenous DNA (SourceTracker2) - Data processing of sample Abusir1630b (M. leprae) -- Read processing, mapping, and variant calling -- SNP typing -- Anthropological analysis -- Phylogeny -- Beast analysis -- Temporal signal - Data processing individual Abusir1543 (hepatitis B virus) -- Read processing, mapping, and variant calling -- Phylogeny -- Recombination analysis -- Beast analysis -- Temporal signal

This study focuses on the multidisciplinary investigation of three stucco-shrouded mummies with mummy portrait from Egypt dating from the late 3rd to the middle of the 4th century AD, corresponding to the late Roman Period. These three mummies were excavated in the early 17th and late 19th centuries in the Saqqara necropolis near the ancient Egyptian capital of Memphis. Two of them experienced an interesting collection history, when they became part of the collection of the Elector of Saxony and King of Poland August II in Dresden, Germany, in 1728. The investigation includes information about the mummies’ discovery, collection history and shroud decoration obtained through Egyptological expertise. In addition, information on the state of preservation, technique of artificial mummification, age at death, sex, body height and health of the deceased was achieved through computed tomography (CT) analysis. Research yielded an adult male, a middle-aged female and a young female. Due to the rather poorly preserved bodies of the male and middle-aged female, a specific technique of artificial mummification could not be ascertained. Brain and several internal organs of the well-preserved young female were identified. Wooden boards, beads of necklaces, a hairpin, and metal dense items, such as lead seals, nails and two coins or medallions were discovered. Paleopathological findings included carious lesions, Schmorl’s nodes, evidence of arthritis and a vertebral hemangioma. The study revealed insights on the decoration and burial preparation of individuals of upper socioeconomic status living in the late Roman Period, as well as comprehensive bioanthropological information of the deceased.

Objective This study was designed to search for ancient parasites in abdominal content and coprolites from Bolivian mummies. Materials Twelve mummified individuals from the Andean highlands, housed at the National Museum of Archaeology (MUNARQ) in La Paz, Bolivia. Methods Microscopic analysis of rehydrated samples (coprolites and abdominal content), following Lutz's spontaneous sedimentation technique. Results Eggs of Enterobius vermicularis were identified in coprolites from one mummy, and capillariid eggs in the organic abdominal content from another individual. Conclusions This is the first evidence of ancient intestinal parasites in Bolivian mummies. Significance This pioneering study focused on the search of ancient intestinal parasites in human remains of the Bolivian Andes and contributes to greater knowledge of paleoparasitology in South America. Limitations All mummies in the MUNARQ belonged to the Andean Bolivian highlands (post-Tiwanaku era or Late Intermediate Period), although the exact provenance of the material and the associated contexts are not well recorded. Suggestions for further research Considering the great number of well-known archaeological sites and other unexplored sites in Bolivia, in addition to large collections in museums, further paleopathological and paleoparasitological molecular studies in mummies and skeletons are called for.

We report, for the first time, a case of nesting by Apis mellifera mellifera (L., 1758) (Insecta: Hymenoptera: Apidae), Vespula vulgaris (L., 1758) (Insecta: Hymenoptera: Vespidae), and Sciurus vulgaris L., 1758 (Mammalia: Rodentia: Sciuridae) inside a mummified human corpse in natural conditions (Lower Silesia province, south-western Poland). A case history, autopsy findings, and biological observations are provided. A list of the insect species found in the corpse is provided and compared with available data. Other zoological findings are mentioned and briefly discussed.

During the restoration of the Saint Antonio Abate Cathedral in Castelsardo, Sardinia, Italy, numerous human remains were found in a crypt. The burial site contained upwards of 120 individuals organized in successive layers from the bottom of the crypt; of these, 18 partially mummified individuals have been identified, including the last 2 individuals buried in the crypt. In the present study, we focused on these 2 individuals by initially adopting a morphologic and anthropological approach. The anthropological analysis of the remains showed that the 2 bodies were partially mummified and that most of the organs were not available; for this reason, the sex was determined by secondary sexual characteristics of the skulls and the long bones. The aim of this research was to describe the general state of the mummified bodies and tissues by morphologic and ultrastructural analysis using light and electron microscopy techniques. To ensure the preservation of specific tissue proteins, immunohistochemical fluorescence analysis was used. Limited information is available regarding the preservation of mummified tissues. Thus, this study demonstrated the presence of muscle and skin tissue markers in a good state of preservation, even though the tissues had undergone a slow mummification process. Our results demonstrate that several types of tissues and cell proteins may survive over a prolonged period and that these materials survive the postmortem processes.

Funerary archaeoentomology entails the study of insects from archaeological contexts, in order to examine funerary practices, thanatology and hygiene/sanitation in ancient populations. However, while insects from human mummies have been widely studied, there is a limited literature dealing with archaeoentomology of animal sacrifices. Andean camelid sacrifices are common in ritual contexts, as funerary or foundation offerings. The current paper addresses camelid remains recovered from the archaeological site of Pachacamac, during the 2016 excavation season. The insect fauna was assessed in order to determine the social context of the remains, and the manner in which the camelids were utilised. The carcasses yielded remains pertaining to Diptera and Coleoptera. The presence of Cochliomyia macellaria (Diptera, Calliphoridae), Sarcophaga sp. (Diptera, Sarcophagidae) and Synthesiomyia nudiseta (Diptera, Muscidae) suggest an initial colonisation in the open, while other species typical of later phases of the colonisation – including Hydrotaea aenescens (Diptera, Muscidae) and members of the family Phoridae – suggest that the carcasses were subsequently buried. Despite the evident importance of camelids to Andean populations, both historically and archaeologically, this is the first time that entomology has been used to examine animal sacrifice methods in this area, and comprises a watershed in the development of multidisciplinary approaches to sacrificial rites in ancient Peru., SCOPUS: ar.j, info:eu-repo/semantics/published

Background The mummified body of a small child was found in a sealed Barstow cast iron casket during construction activity in San Francisco in 2016. Using historical records and ancient DNA the child was determined to be Edith H. Cook. She was born 28 November, 1873 in the city of San Francisco, and died of "marasmus" on 13 October, 1876 also in San Francisco. Aims Currently, there are few techniques for estimating human season of death in archaeological cases. Hydrogen isotope ratios (δ2H) in hair keratin is known to strongly correlate with drinking water. We explore δ2H in serial hair samples as a potential technique to estimate season of death by comparing the δ2H record from hair to the known date of death. Materials & methods Approximately 50 hairs were removed from the scalp, aligned from the root, and cut into 5cm serial sections, each representing approximately 2 weeks of growth, and the total sequence a total of 1 year of growth. δ2H was measured on each 5cm segment and compared to previously-reported δ13C, δ15N, and δ34S values. Results δ2H in the serial hair samples ranged between -56‰ and -48‰, consistent with her water values recorded in surface waters from San Francisco, and follow a sinusoidal pattern. Decreasing δ2H in terminal samples before death suggest Fall as the season of death, consistent with the known date of death. Discussion This test case shows that archaeological hair preserves a seasonal signature in the form of changing keratin δ2H values that correlate to changing δ2H in surface drinking water. Terminal values in root record water ingested during the final week(s) before death. Conclusions We argue that this technique can be used to estimate season of death in future archaeological or forensic cases where hair has been preserved but date of death is unknown.

Indirect measures of cognition have become an important tool in research on addiction. To date, however, no research has examined whether indirect measures of parent attachment relate to substance use. To examine this issue, a sample of college students (N = 121) was asked to complete two measures of explicit attachment (the Relationship Questionnaire; Bartholomew & Horowitz, 1991, and the Adult Attachment Styles; Collins & Read, 1990), and a measure of implicit attachment (the Single Category Implicit Association Test, Karpinski & Steinman, 2006). The indirect attachment measure assessed the strength of automatic mental association between the concepts parents and secure. Participants also completed different measures of tobacco, cannabis, and alcohol use. Results showed that, for most of the participants, the parents were considered a source of security at both the explicit and implicit levels. Direct and indirect attachment measures were not related to each other. Overall, explicit attachment was not related to substance use. However, implicit attachment was significantly associated with the use of licit (tobacco) and illicit (cannabis) drugs. We also found some evidence that polydrug use is especially common among students with an insecure implicit attachment. This is the first study to examine how implicit attachment processes relate to addictive behaviors. The results suggest that implicit attachment, thought to reflect unconscious traces of past experiences, is a better predictor of substance use in college students than direct, self-reported measures of attachment. Further studies should examine whether implicit attachment is associated with severe substance use disorders in clinical populations. (PsycINFO Database Record (c) 2019 APA, all rights reserved).

Intraguild predation (IGP) has been commonly reported between predators and parasitoids used as biological control agents as predators consuming parasitoids within their hosts. However, the effect of parasitoid-mummy consumption on the fitness of the predator and subsequent oviposition site selection have not been well studied. In our study, we conducted two laboratory experiments to examine the influence of Aphidius gifuensis Ashmead (Hymenoptera: Braconidae) mummies as prey on fitness and subsequently oviposition site selection of Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae). Results indicate that when H. axyridis was reared on A. gifuensis mummies only, its larval development was prolonged, and body weight of the 4th instar larvae and newly emerged adults, and fecundity decreased. Moreover, H. axyridis did not exhibit oviposition preference on plants infested with unparasitized aphids or aphids parasitized for shorter than 9 days. However, compared with plants with mummies (parasitized ≥9 days), H. axyridis laid more eggs on plants with unparasitized aphids. In contrast, H. axyridis previously fed with A. gifuensis mummies did not show a significant oviposition preference between plants with unparasitized aphids and those with mummies (parasitized ≥9 days). Overall, our results suggest that mummy consumption reduced the fitness of H. axyridis. Although H. axyridis avoided laying eggs on plants with A. gifuensis mummies, prior feeding experience on A. gifuensis mummies could alter the oviposition site preference. Thus, in biological control practice, prior feeding experience of H. axyridis should be carefully considered for reduction of IGP and increase of fitness of H. axyridis on A. gifuensis.

The general process and pattern of decomposition is well-documented and understood. However, specific environmental conditions may alter this pattern and prematurely terminate the decay process. An example of this is natural mummification – a preservative process characterized by desiccation, brittleness and shrinkage of the skin and body tissues. It is important to understand how, when, and where such variations may occur, and for this reason environmentally-specific studies of decay are required. The aim of the present study was the establish baseline data on soft-tissue decomposition in two terrestrial habitats in the Western Cape. A total of 16 pig carcasses serving as analogues for humans were deployed in these habitats during two successive winters and summers between 2014 and 2016. The rate and pattern of decomposition were assessed via measurement of weight loss over time and scoring the decomposition process using Megyesi et al. (2005) Total Body Score system and study-specific criteria for mummification. Carcasses typically followed the expected pattern of decay with a few exceptions, most notably instances of rapid natural mummification. Natural mummification, as defined by Megyesi et al. (2005), was observed to occur as early as 17 days postmortem, with five carcasses mummifying in less than one month. The timing of natural mummification varies widely, from a few days to several years, averaging around three months in temperate regions. Natural mummification occurring in less than one month is termed precocious mummification and is rarely observed in temperate regions. With only three reports globally, this study’s findings are globally significant, highlighting the importance of regionally-specific decomposition studies. Two local forensic cases wherein precocious mummification has been observed are also presented and, considered together with the study’s results, a possible mechanism driving this process is proposed.

We analyzed Clonorchis sinensis ancient DNA (aDNA) acquired from the specimens of the Joseon mummies. The target regions were cytochrome C oxidase subunit 1 (CO1), internal transcribed spacer 1 (ITS1), nicotinamide adenine dinucleotide hydrogen (NADH) dehydrogenase subunits 2 (NAD2) and 5 (NAD5). The sequences of C. sinensis aDNA was completely or almost identical to modern C. sinensis sequences in GenBank. We also found that ITS1, NAD2 and NAD5 could be good markers for molecular diagnosis between C. sinensis and the other trematode parasite species. The current result could improve our knowledge about genetic history of C. sinensis., Graphical Abstract

Computed tomography studies and histological analyses were performed on the mummified remains found in the Chehrābād salt mine in northwestern Iran. The ancient salt mummies are dated to the Achaemenid (550–330 BC) and Sassanid (3rd–7th century AD) time period and died in mining incidents. The aim of the study was to describe the radiological and histological findings of several ancient Iranian salt mummies with special interest in pathological and postmortem changes. The mummified remains show multiple traumatic alterations, such as fractures and signs of massive compression. Histological analyses can clearly differentiate soft tissue, however the preservation status is variable. These Iranian salt mummies are a rare example of the ancient Iranian population. The soft tissue and organs are well preserved, however in different degrees due to the varying conditions.

Background Cardiovascular disease has emerged as the world's leading cause of death in the last century. An epidemiological focus of this disease that extends not only beyond the developed world but also far back into antiquity asks new questions about associated risk factors. Ancient mummies found in the Atacama desert are well preserved and show signs of cardiovascular disease as early as 1000B.C. in Peru and Chile. Method and results Gross and histopathological examination of specimens shows atherosclerosis, cardiomegaly, endocarditis, and myocardial fibrosis. Conclusion In comparison to other ancient populations, less atherosclerosis has been noted in South American mummies. The chewing of coca leaves, a habitual cultural practice unique to the region, supports evidence of reduced cardiovascular risk among ancient people living in South America.

The study of the gastric pathogen Helicobacter pylori brought us interesting data on the history of mankind. Based on multi-locus sequence typing, it was possible to trace the migration of Homo sapiens all around the world, and to infer the time when he went Out of Africa. Beside these phylogeographic aspects, paleomicrobiology gave us important information on life in the Neolithic period, following the discovery of Ötzi, the Iceman, who was living in the Tyrolean Alps 5200 years ago, and from whom a Helicobacter pylori genome was sequenced. This review presents the data accumulated in these different fields.

Paleopathologists have begun exploring the pathoecology of parasitic diseases in relation to diet and environment. We are summarizing the parasitological findings from a mummy in the site of Lapa do Boquete, a Brazilian cave in the state of Minas Gerais. These findings in context of the archaeology of the site provided insights into the pathoecology of disease transmission in cave and rockshelter environments. We are presenting a description of the site followed by the evidence of hookworm, intestinal fluke, and Trypanosoma infection with resulting Chagas disease in the mummy discovered in the cave. These findings are used to reconstruct the transmission ecology of the site.

The figurative arts and precisely the ancient Pompeian wall paintings portraits can provide an additional source of information in supplementing bio-anthropological studies. There are several genetic diseases with a wide spectrum of congenital bone stigmata in association to distinctive facial features. Gorlin-Goltz syndrome, also named nevoid basal cell carcinoma syndrome, is an autosomal dominant syndrome characterized by unusual skeletal changes, such as macrocephaly, facial asymmetry, hypertelorism, frontal and parietal bossing caused by germline mutations of the gene PTCH1. The Gorlin syndrome, clinically defined in 1963, existed during Dynastic Egyptian times, as revealed by a spectrum of skeletal findings compatible with the syndrome in mummies dating back to three thousand years ago and, most likely, in the ancient population of Pompeii. In the present research, we discuss the potential relationship between Pompeian wall paintings portrait and the cranio-metric bone changes revealed among the Pompeian skull collections assuming that the ancient portraits can constitute an important tool that should be strictly integrated with osteologic and biomolecular data in order to argue a syndromic diagnosis in ancient population.

Understanding parasite history and the evolution of host/parasite relationships is one of the most important aspects of paleoparasitology. Within the framework of this research topic, this paper focuses on the human pathogenic amoeba, Entamoeba histolytica. The compilation of all the available archaeological data concerning this parasite leads to a first glimpse of the history of this parasite of current medical importance. Paleoparasitological investigation into this parasite uses immunological techniques and shows that the modern strain of E. histolytica has been present in Western Europe since at least the Neolithic period (3700yearsBCE), and could have originated in the Old World. The appearance of the modern amoeba strain in the pre-Columbian Americas and the Middle East around the 12th century CE gives rise to hypotheses as to how human migrations (Atlantic or Pacific routes) contributed to the diffusion of this pathogen, resulting in its current distribution. This compilation proves that parasites are valuable proxies for studying past human and animal migrations, and should be given more consideration in the future.

The Guanches, ancient inhabitants of the Canary Islands, Spain, practiced mummification of their dead. A paleoparasitological and paleogenetic analysis was conducted on mummified bodies (n = 6) (AD 1200, Cal BP 750) belonging to the Guanche culture from Gran Canaria Island. Coprolite and sediment samples (n = 19) were removed from below the abdominal region or sacral foramina. The samples were rehydrated in 0.5% trisodium phosphate solution for 72 hr at 4 C, and the paleoparasitological investigation was conducted by spontaneous sedimentation method and microscopic examination. The results revealed the presence of well-preserved eggs of Ascaris sp., Trichuris trichiura , Enterobius vermicularis , and hookworms. Ancient DNA was extracted from sediment samples to elucidate the ancestry of the mummies and for molecular detection of Ascaris sp. infection. Results of paleogenetic analysis demonstrated Ascaris sp. infection using 2 molecular targets, cytb and nad1. The mtDNA haplotypes U6b, U6b1, and HV were identified, which confirmed records of Guanche ancestry. The excellent preservation of Guanche mummies facilitated the paleoparasitological and paleogenetic study, the results of which contribute to our knowledge of Guanche culture and their health status.

Tuberculosis (TB) was a large burden of infections that peaked during the 19th century in Europe. Mummies from the 18th century CE, discovered in the crypt of a church at Vac, Hungary, had high TB prevalence, as revealed by amplification of key fragments of TB DNA and genome-wide TB analysis. Complementary methods are needed to confirm these diagnoses and one approach uses the identification of specific lipid biomarkers, such as TB mycocerosic acids (MCs). Previously, MC derivatives were profiled by specialised gas chromatography-mass spectrometry (GC-MS), so an alternative more direct approach has been developed. Underivatized MCs are extracted and analysed by high-performance liquid chromatography linked to a mass spectrometer, in heated electrospray ionisation mode (HPLC-HESI-MS). The method was validated using representatives of the Mycobacterium tuberculosis complex and other mycobacteria and tested on six Vac mummy cases, previously considered positive for TB infection. Analysing both rib and soft tissue samples, four out of six cases gave profiles of main C32 and major C29 and C39 mycocerosates correlating well with those of M. tuberculosis. Multidisciplinary methods are needed in the diagnosis of ancient tuberculosis; this new protocol accesses important confirmatory evidence, as demonstrated by the confirmation of TB in the Vac mummies.

Human ectoparasites, including lice, have been recovered from a wide range of archaeological materials. The human head louse, Pediculus humanus capitis, has been identified from mummies and sediments for decades. Louse eggs are the body part most commonly encountered and therefore the most frequently quantified. Typically, several types of microscopy are applied for egg documentation. For studies in which quantification of infestation is a goal, counting is done with the naked eye or with the aid of handheld lenses. For determination and stage classification, stereomicroscopy is commonly used. For more detailed examination of microstructure, light microscopy, scanning electron microscopy (SEM), and confocal laser scanning microscopy (CLSM) can be employed. In most reports, researchers use two or more techniques to accomplish interrelated goals. Automontage microscopy is used to document prehistoric arthropods with good success. Herein, we report the results of a combination of SEM and automontage microscopy to document lice and eggs recovered from South American mummies. This combined approach allows for simultaneous examination of internal and external characteristics. Thirty automontage composite images of 2 adult lice and 16 eggs showed that egg internal morphologies were easily examined showing the within-egg anatomy of emergent nymphs. SEM imaging of 9 lice and 129 eggs was completed. In the case of two adults and several eggs, SEM imaging was accomplish after automontage image capture of the same specimens. This one-to-one image comparison of SEM and automontage shows that transmitted light of automontage reveals egg internal structures and details of the adult lice. SEM allows for high magnification examination of egg, nymph and adult microstructures. We conclude that automontage imaging followed by SEM results in efficient graphic documentation of rare louse specimens.

The genetic information of ancient Paragonimus westermani, the oriental lung fluke infecting over 20 million people worldwide, has not been thoroughly investigated thus far. We analysed genetic markers (COI and ITS2) of P. westermani from coprolite specimens (n = 6) obtained from 15th to 18th century Korean mummies. Our results indicated that all P. westermani sequences were generally distinct from the other species of the genus Paragonimus. The sequences were clustered into three groups: Group I for East Asia; Group II for South and Southeast Asia; and Group III for India and Sri Lanka. In this study, we found that ancient P. westermani sequences in Korea belong to Group I, adding invaluable information to the existing knowledge of Paragonimus paleogenetics.

The ancient catacombs of Egypt harbor millions of well-preserved mummified Sacred Ibis (Threskiornis aethiopicus) dating from ∼600BC. Although it is known that a very large number of these ‘votive’ mummies were sacrificed to the Egyptian God Thoth, how the ancient Egyptians obtained millions of these birds for mummification remains unresolved. Ancient Egyptian textual evidences suggest they may have been raised in dedicated large-scale farms. To investigate the most likely method used by the priests to secure birds for mummification, we report the first study of complete mitochondrial genomes of 14 Sacred Ibis mummies interred ∼2500 years ago. We analysed and compared the mitogenomic diversity among Sacred Ibis mummies to that found in modern Sacred Ibis populations from throughout Africa. The ancient birds show a high level of genetic variation comparable to that identified in modern African populations, contrary to the suggestion in ancient hieroglyphics (or ancient writings) of centralized industrial scale farming of sacrificial birds. This suggests a sustained short-term taming of the wild Sacred Ibis for the ritual yearly demand.

The impressive Sacristy of the Basilica of San Domenico Maggiore contains 38 wooden sarcophagi with the bodies of 10 Aragonese princes and other Neapolitan nobles, who died in the 15th and 16th centuries. To improve the knowledge about the entomofauna associated with bodies in archaeological contexts, herein we provide insights on the funerary practices and the insect community associated to Ferrante II King of Naples and other Italian Renaissance mummies of the Aragonese dynasty buried in the Basilica of St. Domenico Maggiore. We identified 842 insect specimens: 88% were Diptera (Muscidae, Fanniidae, and Phoridae), followed by 9% Lepidoptera (Tineidae) and 3% Coleoptera (Dermestidae and Ptinidae). Ninety-seven percent of the specimens were collected from the coffin of Francesco Ferdinando d’Avalos, which was the best preserved. A lack of fly species characterizing the first colonization waves of exposed bodies was noted. The most common fly was the later colonizing muscid Hydrotaea capensis (Wiedemann); only a few Fanniidae (Fannia spp.) were retrieved. The lack of blowflies, coupled with recording H. capensis as the dominant fly, supports our hypothesis that corpses have been kept indoors for a long time under confined environmental conditions. Other explanations include odorous oils/balms having been used in the embalming process, causing the delay or stopping the arrival of first colonizer flies. Hermetically sealing of the coffin with bitumen may also have played a role in preventing access to the corpses. This scenario describes a historical context characterized by a well-advanced knowledge of body preparation, with specific burial techniques adopted for nobles.

In 1798, Napoleon Bonaparte's army invaded Egypt, returning with many treasures including large numbers of Sacred Ibis mummies. The ancient Egyptians revered the ibis and mummified literally millions of them. The French naturalist Georges Cuvier used these mummies to challenge an emerging idea of the time, namely Jean-Baptiste Lamarck's theory of evolution. Cuvier detected no measurable differences between mummified Sacred Ibis and contemporary specimens of the same species. Consequently, he argued that this was evidence for the "fixity of species." The "Sacred Ibis debate" predates the so-called "Great Debate" between Cuvier and Geoffroy Saint-Hilaire and the publication of Darwin's On the Origin of Species five decades later. Cuvier's views and his study had a profound influence on the scientific and public perception of evolution, setting back the acceptance of evolutionary theory in Europe for decades.

In Ancient Egypt it was common practice to embalm corpses and specific internal organs to ensure eternal life. The exact nature of the employed embalming fluids, particularly for organ preservation within the canopic jars, is debated. Therefore, the aim of the current study, was to chemically characterize and differentiate canopic jars (n = 28) and mummies (n = 6) using gas chromatography - high resolution mass spectrometry (GC-HR MS) with a new untargeted metabolomics-like screening approach; as part of a larger minimal-invasive transdisciplinary study on Ancient Egyptian human tissues. Post-analytical data processing included deconvolution, screening against the NIST 14 spectral database as well as a high resolution metabolomics library, and positive peak evaluation. In the majority of samples the presence of a coniferous resin was indicated by the detection of longiborneol in combination with abietadiene acid derivatives and guajacol. Beeswax, proposedly used for symbolic reasons and/or as a binding agent, was detected in 10 samples. Previously not mentioned in the literature, but identified in the current sample set, were medical-related substances like aniseed constituents, salicylic acid, chamazulene and jacobine. By applying an untargeted metabolomics-like approach to archaeological samples for the first time, extensive statistical analysis was made possible (using both identified and non-identified features; adding up to 4381 features), which showed significant differences in the overall chemical composition of canopic jar and mummy samples using principle component analysis (PCA) and partial least square-discriminant analysis (PLS-DA). This emphasizes the necessity for more extensive canopic jar studies in the future in order to interpret findings correctly.

Summary The history of humankind is marked by the constant adoption of new dietary habits affecting human physiology, metabolism, and even the development of nutrition-related disorders. Despite clear archaeological evidence for the shift from hunter-gatherer lifestyle to agriculture in Neolithic Europe [1], very little information exists on the daily dietary habits of our ancestors. By undertaking a complementary -omics approach combined with microscopy, we analyzed the stomach content of the Iceman, a 5,300-year-old European glacier mummy [2, 3]. He seems to have had a remarkably high proportion of fat in his diet, supplemented with fresh or dried wild meat, cereals, and traces of toxic bracken. Our multipronged approach provides unprecedented analytical depth, deciphering the nutritional habit, meal composition, and food-processing methods of this Copper Age individual., Highlights • The last meal of the Iceman, a European Copper Age mummy, was reconstructed • Our multipronged approach deciphers the meal composition and food processing • His high-fat diet was supplemented with wild meat and cereals, Maixner et al. report the dietary reconstruction of the Iceman’s last meal using a combined multi-omics approach. The stomach content analysis of the 5,300-year-old glacier mummy shows that the Iceman’s diet preceding his death was a mix of carbohydrates, proteins, and lipids, well adjusted to the energetic requirements of his high-altitude trekking.

In this article, a complete history of Charlemagne's mortal remains is outlined, including the first publication reporting on the most recent tomb opening in 1988. Besides exclusive bioarchaeological details - namely that his body was indeed mummified - a full clinical interpretation of the Emperor's final illnesses and death is given: a likely combination of osteoarthritis, gout and a recurrent fever caused by an infectious disease.

The preservation of the meningeal artery in ancient mummified bodies, particularly in anthropogenic Egyptian mummies, is a highly controversial topic in neuroscience and anthropological research. A recent (2015) debate between Wade and Isidro, based on the interpretation of the meningeal grooves and cast in a skull from the necropolis of Kom al-Ahmar Sharuna (Egypt), highlighted both the necessity of having clear radiological descriptions of this anatomical structure and of assessing large collections of mummified crania. Here, we present for the first time an instance of extremely well-preserved middle meningeal artery in the mummy of the ancient Egyptian dignitary Nakht-ta-Netjeret (ca. 950 BC), still inside the meninx, using paleo-radiological techniques. We finally link this find with experimental data from the neurological literature on the methodology of excerebration implemented by ancient Egyptian embalmers.

The Bioarchaeology of Care approach developed by Tilley is usually applied to skeletalized human remains, given the usual constraints of preservation bias that are seen with archaeological assemblages. However, other tissues, such as hair are sometimes preserved and can provide a wealth of information that can supplement the skeletal data. Archaeological hair has been analysed for drug compounds for almost thirty years. This article integrates data from hair analyses for coca metabolites, stable light isotope analysis and aDNA to expand the potential of the Bioarchaeology of Care approach using the example of a spontaneously mummified adult female from northern Chile.

Tuberculosis has affected Europe for millennia and continues to be a burden upon modern society. It is estimated that one-third of the world's population is infected with Mycobacterium tuberculosis, the causative agent of this condition. Despite the introduction of control strategies, the disease continues to be one of the most common causes of death globally. Within the framework of the Lithuanian Mummy Project, seven spontaneously mummified human bodies from a church crypt in Vilnius, dating from the 18th and 19th century, were CT-scanned to assess the presence of tuberculosis or other lung diseases. We encountered pulmonary lesions suggestive of cases of pulmonary tuberculosis. In addition, one case might have been affected by extra-pulmonary tuberculosis. This report replicates the image findings from previous studies on ancient mummies that provided evidence of tuberculosis in soft tissues, thus helping reconstruct the history of this disease over time.

Objective: To study pathologies, peri- and postmortal alterations as well as the general preservation state of an ancient Iranian salt mummy. Materials and methods: Several mummified remains from two different time periods (1500-2500 BP) were found in the Chehrābād salt mine in Iran. Computed tomography was performed on Salt Man #4 (410-350 BC), the best preserved out of the six salt mummies (Siemens, Sensation 16; 512 × 512 matrix; 0.75-5mm slice thickness, 240-mA tube current, 120-kV tube voltage, and 0.976-mmpixel size). Results: Radiological analyses showed an excellent state of preservation of an adolescent body. Several normal variants such as aplasia of the frontal sinus as well as a rare congenital deformation of the 5th vertebral body (butterfly vertebra) have been observed. The individual shows multiple fractures, which is consistent with the theory that he died due to a collapse in the ancient salt mine. Conclusions: The salt preserved the soft tissue as well as parts of the inner organs remarkably well. However, further investigations including histology are needed to reveal additional details of the health status of this unique salt mummy.

Observations of animals' responses to dying or dead companions raise questions about their awareness of states of helplessness or death of other individuals. In this context, we report the case of a female Tonkean macaque (Macaca tonkeana) that transported the body of her dead infant for 25 days and cannibalized its mummified parts. The mother appeared agitated in the first 2 days after the birth. She then took care of her infant's corpse, which progressively dried and became mummified. In a third stage, the mother continued to transport the corpse as it started disintegrating, and she gnawed and consumed some parts of the remains. Our observations suggest that mummification of the body favored persistence of maternal behaviors by preserving the body's shape. The female gradually proceeded from strong attachment to the infant's body to decreased attachment, then finally full abandonment of the remains.

Little is still known about the microbiome resulting from the process of mummification of the human gut. In the present study, the gut microbiota, genes associated with metabolism, and putative resistome of Inca and Italian nobility mummies were characterized by using high-throughput sequencing. The Italian nobility mummies exhibited a higher bacterial diversity as compared to the Inca mummies when using 16S ribosomal (rRNA) gene amplicon sequencing, but both groups showed bacterial and fungal taxa when using shotgun metagenomic sequencing that may resemble both the thanatomicrobiome and extant human gut microbiomes. Identification of sequences associated with plants, animals, and carbohydrate-active enzymes (CAZymes) may provide further insights into the dietary habits of Inca and Italian nobility mummies. Putative antibiotic-resistance genes in the Inca and Italian nobility mummies support a human gut resistome prior to the antibiotic therapy era. The higher proportion of putative antibiotic-resistance genes in the Inca compared to Italian nobility mummies may support the hypotheses that a greater exposure to the environment may result in a greater acquisition of antibiotic-resistance genes. The present study adds knowledge of the microbiome resulting from the process of mummification of the human gut, insights of ancient dietary habits, and the preserved putative human gut resistome prior the antibiotic therapy era.