Biomolecular analyses enable new insights into ancient Egyptian embalming.

The ability of the ancient Egyptians to preserve the human body through embalming has not only fascinated people since antiquity, but also has always raised the question of how this outstanding chemical and ritual process was practically achieved. Here we integrate archaeological, philological and organic residue analyses, shedding new light on the practice and economy of embalming in ancient Egypt. We analysed the organic contents of 31 ceramic vessels recovered from a 26th Dynasty embalming workshop at Saqqara1,2. These vessels were labelled according to their content and/or use, enabling us to correlate organic substances with their Egyptian names and specific embalming practices. We identified specific mixtures of fragrant or antiseptic oils, tars and resins that were used to embalm the head and treat the wrappings using gas chromatography-mass spectrometry analyses. Our study of the Saqqara workshop extends interpretations from a micro-level analysis highlighting the socio-economic status of a tomb owner3-7 to macro-level interpretations of the society. The identification of non-local organic substances enables the reconstruction of trade networks that provided ancient Egyptian embalmers with the substances required for mummification. This extensive demand for foreign products promoted trade both within the Mediterranean8-10 (for example, Pistacia and conifer by-products) and with tropical forest regions (for example, dammar and elemi). Additionally, we show that at Saqqara, antiu and sefet-well known from ancient texts and usually translated as 'myrrh' or 'incense'11-13 and 'a sacred oil'13,14-refer to a coniferous oils-or-tars-based mixture and an unguent with plant additives, respectively.

Pot residues show how ancient Egyptians made a mummy.

Analysis reveals the chemicals applied to corpses in a 2700-year-old mummification workshop.

Alfredo Salafia's handwritten memoir and the embalming of Rosalia Lombardo: a commentary.

Alfredo Salafia, an Italian embalmer who was active in the early 20th century, treated a number of corpses with his own preservation method. Among them was the young Rosalia Lombardo, just shy of two years old at the time of her death, whose remarkable preservation has brought her significant attention. It is well documented that Salafia had concocted a formula for a highly advanced embalming fluid which is believed to have been used to preserve Lombardo. This is referenced in a book and in two unpublished documents, and was also traced through familial memories and stories within the Capuchin Brotherhood of Palermo, where three of those bodies embalmed by Salafia are kept in a world-renowned crypt. Salafia's paternity of this specific preparation is further supported by some indirect evidence, which corresponds to what the embalmer described in his handwritten memoir. A recent article by Galassi and co-workers, however, attempted to debunk Salafia's role, and questioned the results of previous research performed on this case. They suggest additional studies on Lombardo that are not only unnecessary, but would irreversibly damage the long-term conservation of the mummy. Our paper aims to demonstrate that their article is biased and built upon incorrect assumptions and interpretations, thus re-establishing the validity of the narrative shared up until now.

Generalized dermatitis in the natural mummy of the Roman Catholic nun Marie-Léonie Martin (France, 1863-1941).

OBJECTIVE: To document skin lesions on a mummified individual from the XIX century and to diagnose the pathology based historical documentation and physical examination. MATERIALS: Marie Leonie Martin (1863-1941) was a Roman Catholic nun. Her naturally mummified body is currently preserved in the Monastery of the Visitation in Caen (France). On the occasion of her beatification, the body was exhumed, studied and restored for conservation purposes. METHODS: The mummy was analyzed histologically and with CT imaging. RESULTS: The examination of the body noted areas of skin discoloration of reddish color; the paleopathological investigations revealed the presence of skin lesions (pustules) distributed throughout the body, with the exception of the face, abdomen and palms. The histological analyses of the pustules showed the presence of a central duct for the leakage of exudate, hyperkeratosis of the stratum corneum and a proliferation of cells in the spinous layer (acanthosis) of the epidermis. CONCLUSIONS: The red discoloration of the skin is typical of extensive erythema. SIGNIFICANCE: Our understanding of skin conditions in the past is extremely limited. This study provides the first paleopathological case of diagnosed unclassified endogenous eczema and provides insight into the antiquity and effects of the disease. LIMITATIONS: It was not possible to carry out a genetic analysis due to DNA contamination of the mummy.

Multidisciplinary investigation of an ancient renal stone in a mummy from Popoli, central Italy.

The renal stone found in the natural mummy of an anonymous nobleman dating to 19th century was investigated using advanced imaging modalities and analytic investigations. By this multidisciplinary approach we were able to identify the chemical components and their distribution throughout the sample. These results allowed to understand the lifestyle habits of the subject, as well as the exact pathogenesis of his disease.

The importance of the nasopharynx and anterior skull base in excerebration techniques from KV40, a New Kingdom Egyptian site.

In ancient Egypt, a unique technique for removing the brain was invented as part of the mummification practice and refined over the centuries. This usually involved piercing the anterior skull base through a nasal passage to remove the brain remnants through that perforation. From 2010 to 2018, an interdisciplinary team of the Universities of Basel and Zurich investigated tomb no. 40 (KV40) in the Valley of the Kings, Luxor, Egypt. Archaeological findings indicate a first burial phase during the mid-18th Dynasty (ca. 1400-1350 BCE) and a second in the 22nd to 25th Dynasty (approx. 900-700 BCE). Repeated looting since ancient times severely damaged and commingled the human remains of the two burial phases. The detailed examination of the skulls showed evidence of different transnasal craniotomy practices. This study aims to provide a systematic presentation of the evidence for different excerebration techniques found in the mummy heads, skulls, and skull fragments from KV40, reflecting the long period of occupancy of this tomb by individuals of different social classes.

The genomic origins of the Bronze Age Tarim Basin mummies.

The identity of the earliest inhabitants of Xinjiang, in the heart of Inner Asia, and the languages that they spoke have long been debated and remain contentious1. Here we present genomic data from 5 individuals dating to around 3000-2800 BC from the Dzungarian Basin and 13 individuals dating to around 2100-1700 BC from the Tarim Basin, representing the earliest yet discovered human remains from North and South Xinjiang, respectively. We find that the Early Bronze Age Dzungarian individuals exhibit a predominantly Afanasievo ancestry with an additional local contribution, and the Early-Middle Bronze Age Tarim individuals contain only a local ancestry. The Tarim individuals from the site of Xiaohe further exhibit strong evidence of milk proteins in their dental calculus, indicating a reliance on dairy pastoralism at the site since its founding. Our results do not support previous hypotheses for the origin of the Tarim mummies, who were argued to be Proto-Tocharian-speaking pastoralists descended from the Afanasievo1,2 or to have originated among the Bactria-Margiana Archaeological Complex3 or Inner Asian Mountain Corridor cultures4. Instead, although Tocharian may have been plausibly introduced to the Dzungarian Basin by Afanasievo migrants during the Early Bronze Age, we find that the earliest Tarim Basin cultures appear to have arisen from a genetically isolated local population that adopted neighbouring pastoralist and agriculturalist practices, which allowed them to settle and thrive along the shifting riverine oases of the Taklamakan Desert.

The Oshu Fujiwara-An interdisciplinary study on the history, culture and medical assessment of the oldest known mummified human remains in Japan (late Heian, 12th century AD).

This study documents a rare case of mummified human remains from Japan, dating to the late Heian period, 12th Century AD. The remains have only been scientifically investigated once in 1950 so far. The results of this investigation were translated, analyzed, and interpreted using methods of the 21st century. The remains have been traditionally identified as the four ruling generations of the Oshu Fujiwara clan, who built a cultural and economic center in Hiraizumi. Accordingly, this paper will first examine the historical and cultural significance of Hiraizumi and its ruling class before re-evaluating the findings of the 1950 investigation. This study is the first in the Western scientific literature to provide a comprehensive historical, cultural, and medical evaluation of these mummies.

Giant cell tumor of bone in an eighteenth-century Italian mummy.

Giant cell tumor (GCT) of the bone is a locally aggressive and rarely metastasizing neoplasm. It is composed of neoplastic mononuclear stromal cells with a monotonous appearance admixed with macrophages and osteoclast-like giant cells. In a small subset of cases, GCT is malignant. Terminology previously related to this entity, and which is no longer supported by the World Health Organization, includes osteoclastoma and benign fibrous histiocytoma (BFH). Giant cells occur in numerous other pathologic conditions of the bone, which accounts for the misrepresentation of these non-GCT tumors in the early literature. Non-ossifying fibroma (NOF), aneurysmal bone cyst, and chondroblastoma have been erroneously labeled GCT for this reason. A single description of an ancient GCT was reported by Brothwell and Sandison and subsequently mentioned by Aufderheide and Rodrìguez-Martìn who were astonished that more of these tumors had not been identified in archaeological cases. To the best of our knowledge, no other cases of ancient GCT have been cited in the paleopathology literature. The study of this type of neoplasm in antiquity can be used as a means to better understand its characteristics and behavior and to expand the depth of time of the etiology of these lesions. We report a case of GCT of the left femur observed following the total body CT imaging of a partially mummified adult female, dating to eighteenth century.

Radiological and histological findings in ancient salt mummies from the salt mine of Douzlakh, Iran.

Computed tomography studies and histological analyses were performed on the mummified remains found in the Chehrabad 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.

Molecular analysis of black coatings and anointing fluids from ancient Egyptian coffins, mummy cases, and funerary objects.

Black organic coatings and ritual deposits on ancient Egyptian coffins and cartonnage cases are important and understudied sources of evidence about the rituals of funerary practice. Sometimes, the coatings were applied extensively over the surface of the coffin, resembling paint; in other cases, they were poured over the mummy case or wrapped body, presumably as part of a funerary ritual. For this study, multiple samples of black coatings and ritual liquids were taken from 20 Egyptian funerary items dating to a specific time period (c. 943 to 716 BC). Multiple sampling from each object enabled several comparisons to be made: the variability of the black coating within one application, the variability between two applications on one object, and the variability from object to object. All samples were analyzed for lipids using gas chromatography-mass spectrometry (GC-MS), and 51 samples from across the 20 items were further analyzed for the presence of bitumen using solid phase separation followed by selected ion monitoring GC-MS. The majority of the black substances were found to comprise a complex mixture of organic materials, including bitumen from the Dead Sea, conifer resin, and Pistacia resin, providing evidence for a continuation in international trade between Egypt and the eastern Mediterranean after the Late Bronze Age. Both the coating and the anointing liquid are very similar to mummification balms, pointing to parallels with Egyptian embalming rituals and raising questions about the practical aspects of Egyptian funerary practice.

Insights from ancient DNA analysis of Egyptian human mummies: clues to disease and kinship.

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.

Five thousand years of bellyaches: Exploring boron concentration in ancient populations of the Atacama Desert.

OBJECTIVES: This study explores whether ancient Atacama Desert populations in northern Chile were exposed to endemic boron contamination. MATERIALS AND METHODS: Using Laser Ablation-Inductively Coupled Plasma-Mass Spectrometry (LA-ICP-MS), we studied 144 strands of ancient mummy hair, ranging from 3000 B.C. to 1500 A.D., excavated from the Lluta, Azapa, and Camarones valleys in northern Chile. We tested whether these ancient populations showed signs of significant boron concentration in hair tissue. RESULTS: On average, all individuals from these valleys showed high boron concentrations, ranging from 1.5 to 4 times above the average boron concentration in contemporary hair (baseline <0.85 µg/g). The boron concentration in mummy hair varied according to the main geographic areas mentioned above. CONCLUSIONS: The rivers of northern Chile have high geogenic boron concentrations. They contain 38 times above the recommended limit for human consumption. Geogenic boron contamination likely played a role in population morbidity and the types of crops that were cultivated in antiquity. The ancient populations were chronically affected by boron overexposure, suggesting that ancient geogenic water contamination should be considered when discussing the biocultural trajectories of ancient populations.

Verification of tuberculosis infection among Vác mummies (18th century CE, Hungary) based on lipid biomarker profiling with a new HPLC-HESI-MS approach.

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 Vác, 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 Vác 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 Vác mummies.

Metagonimus yokogawai Ancient DNA Recovered from 16th- to 17th-Century Korean Mummy Feces of the Joseon Dynasty.

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.

Mummified baboons reveal the far reach of early Egyptian mariners.

The Red Sea was witness to important events during human history, including the first long steps in a trade network (the spice route) that would drive maritime technology and shape geopolitical fortunes for thousands of years. Punt was a pivotal early node in the rise of this enterprise, serving as an important emporium for luxury goods, including sacred baboons (Papio hamadryas), but its location is disputed. Here, we use geospatial variation in the oxygen and strontium isotope ratios of 155 baboons from 77 locations to estimate the geoprovenance of mummified baboons recovered from ancient Egyptian temples and tombs. Five Ptolemaic specimens of P. anubis (404-40 BC) showed evidence of long-term residency in Egypt prior to mummification, consistent with a captive breeding program. Two New Kingdom specimens of P. hamadryas were sourced to a region that encompasses much of present-day Ethiopia, Eritrea, and Djibouti, and portions of Somalia and Yemen. This result is a testament to the tremendous reach of Egyptian seafaring during the 2nd millennium BC. It also corroborates the balance of scholarly conjecture on the location of Punt.

Strontium is a chemical element that can act as a geographic fingerprint: its composition differs between locations, and as it enters the food chain, it can help to retrace the life history of extant or past animals. In particular, strontium in teeth ­ which stop to develop early ­ can reveal where an individual was born; strontium in bone and hair, on the other hand, can show where it lived just before death. Together, these analyses may hold the key to archaeological mysteries, such as the location of a long-lost kingdom revered by ancient Egyptians. For hundreds of years, the Land of Punt was one of Egypt's strongest trading partners, and a place from which to import premium incense and prized monkeys. Travellers could reach Punt by venturing south and east of Egypt, suggesting that the kingdom occupied the southern Red Sea region. Yet its exact location is still highly debated. To investigate, Dominy et al. examined the mummies of baboons present in ancient Egyptian tombs, and compared the strontium compositions of the bones, hair and teeth of these remains with the ones found in baboons living in various regions across Africa. This shed a light on the origins of the ancient baboons: while some were probably raised in captivity in Egypt, others were born in modern Ethiopia, Eritrea, Djibouti, Somalia and Yemen ­ areas already highlighted as potential locations for the Land of Punt. The work by Dominy et al. helps to better understand the ancient trade routes that shaped geopolitical fortunes for millennia. It also highlights the need for further archaeological research in Eritrea and Somalia, two areas which are currently understudied.

The infant mummy's face-Paleoradiological investigation and comparison between facial reconstruction and mummy portrait of a Roman-period Egyptian child.

In Graeco-Roman times in the Lower-Egyptian Fayoum region, a painted portrait was traditionally placed over the face of a deceased individual. These mummy portraits show considerable inter-individual diversity. This suggests that those portraits were created separately for each individual. In the present study, we investigated a completely wrapped young infant mummy with a typical mummy portrait by whole body CT analysis. This was used to obtain physical information on the infant and provided the basis for a virtual face reconstruction in order to compare it to the mummy portrait. We identified the mummy as a 3-4 years old male infant that had been prepared according to the typical ancient Egyptian mummification rites. It most probably suffered from a right-sided pulmonary infection which may also be the cause of death. The reconstructed face showed considerable similarities to the portrait, confirming the portrait's specificity to this individual. However, there are some differences between portrait and face. The portrait seems to show a slightly older individual which may be due to artistic conventions of that period.

First report in pre-Columbian mummies from Bolivia of Enterobius vermicularis infection and capillariid eggs: A contribution to Paleoparasitology studies.

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.

G. B. Morgagni Among Human Pathology, Forensic Medicine and Mummiology. The Beatification of Gregorio Barbarigo of Padua

The article is the first step of a research project aimed at investigating new perspectives and aspects of Morgagni's role and work. His activities as a medical examiner and forensic doctor are yet to be truly discovered. Manuscripts, written by Morgagni when he was a forensic expert for the Health Magistrate of Venice, currently preserved at the City Library in Forli (Italy), shed light on a new aspect of his cultural background. As a forensic doctor, he also helped push an increase in "social medicine" in Italy, when physicians began to collaborate with the administrative and political institutions in order to plan environmental and urban regulations to control air quality. While reading his reports, his contribution to the primordial medical Hygiene and Public Health emerges. Among his reports, the authors focused on the one concerning the Beatification of Gregorio Barbarigo, which clearly highlights his pathological approach, as well as his knowledge and application of embalming systems and mummiology. Moreover, this report could be considered as an issue in the history of paleopathology.