Identification of Diptera Puparia in Forensic and Archeo-Funerary Contexts.

Diptera identification is fundamental in forensic entomology as well as in funerary archeoentomology, where the challenge is exacerbated by the presence of immature stages such as larvae and puparia. In these two developmental stages, specimens possess a very limited number of diagnostic features, and for puparia, there is also a lack of identification tools such as descriptions and identification keys. Morphological analysis, DNA-based techniques, and cuticular chemical analyses all show good potential for species identification; however, they also have some limitations. DNA-based identification is primarily hindered by the incompleteness of genetic databases and the presence of PCR inhibitors often co-extracted from the puparial cuticle. Chemical analysis of the cuticle is showing promising results, but this approach is also limited by the insufficient profile database and requires specific, expensive equipment, as well as trained personnel. Additionally, to ensure the repeatability of the analysis-a critical aspect in forensic investigations-and to preserve precious and unique specimens from museum collections, non-invasive protocols and techniques must be prioritized for species identification.

The history of the Vienna Protocol on dealing with Holocaust era human remains and its resonance with ethical considerations in African American bioarcheology.

The Vienna Protocol on How to Deal with Holocaust Era Human Remains describes what to do when possibly Jewish human remains are found. Based on Jewish medical ethics, it responds to the 2014-2017 discoveries of human remains stemming from biomedical contexts of the Nazi period. Among the finding sites were the Dahlem campus of the Free University of Berlin, the Medical University of Strasbourg, and Max Planck Institute archives. The Vienna Protocol is unique among similar recommendations on Nazi era human remains in its representation of the voices of those who suffered violence and were targeted as victims by Nazi persecution. In addition to discussing the ethics of dealing with physical human remains, these recommendations address the use of images (i.e., visual data) from the bodies of victims of Nazi violence. This paper presents the historical background of the Vienna Protocol and its impact. It also offers a first analysis as to why, at the time of the protocol's formulation, its authors were unaware of its resonance with ethical considerations from African American bioarcheology and a new ethical culture in bioanthropology. Potential reasons for this disconnect may include the historic marginalization of the voices of black scholars in anthropology within the wider scientific community. However, more detailed studies are needed to analyze similarities and differences between the histories and continuities of antisemitism and racism in Europe and the U.S.A., and their ties with scientific theories and practices of disciplines that gain knowledge from human remains.

Female biased adult sex ratio in the Bronze Age cemetery of Shahr-i Sokhta (Iran) as an indicator of long distance trade and matrilocality.

OBJECTIVES: This paper starts from the unusual observation of the overrepresentation of females among adults in the cemetery of Bronze Age Shahr-i Sokhta (Seistan, Iran) and explores the post marital residence pattern. By integrating taphonomical (skeletal preservation), anthropological (sex ratio [SR], sexual dimorphism, stress indicators, age at death), archeological (long distance trade indicators, habitation floor area, social role of women), and ancient DNA (heterozygosity levels in X chromosomes) data we test the hypothesis of post marital matrilocality in the site. METHODS: We computed the SR (pelvis-based sex determination) in a random unpublished adult sample from the cemetery of Shahr-i Sokhta and in two samples previously published by other authors. We used comparative data on SR from: a large Supra Regional multi-chronological sample of sites, n = 47, with 8808 adult sexed individuals, from Southern Europe, Egypt, Middle East, Southern Russia; a Regional Bronze Age sample of sites (n = 10) from Bactria Margiana and Indus Valley with 1324 adult sexed individuals. We estimated the heterozygosity levels in X chromosomes compared with the rest of the autosomes on the assumption that in a matrilocal society females should show lower variability than men. RESULTS: Adult SR in a sample (n = 549) from Shahr-i Sokhta is 70.5, the overrepresentation of females is shared with Regional Bronze Age sites from Bactria Margiana (SR = 72.09) and Indus Valley (SR = 67.54). On the contrary, in a larger Supra Regional multi-chronological sample of sites, mean SR ranges between 112.7 (Bronze Age) and 163.1 (Middle Ages). Taphonomical and anthropological indicators do not explain the overrepresentation of female skeletons. Archeological indicators suggest a high social status of women and that the society was devoted to long range trade activities. heterozygosity levels in X chromosomes are in agreement with a matrilocal society. CONCLUSIONS: Indicators suggest that Bronze Age Shahr-ì Sokhta was a matrilocal society and that long distance trade was an important economic factor producing an overrepresentation of adult female skeletons in the cemetery.

"Heal the sick": Health status and caregiving during the 17th-18th century in Northern Italy (St. Biagio cemetery, Ravenna).

OBJECTIVE: The study of health-related care provision in archeology gives important indications on the culture and community organization of past populations. This study aims to assess the health status of the skeletal assemblage recovered from the burial site of St. Biagio (Ravenna, 17th-18th Centuries); next, we identified likely instances of need for and receipt of caregiving in response to the condition, to examine evidence of community attitudes toward disease and disability. MATERIALS AND METHODS: The skeletal assemblage is composed of 133 individuals in a generally good state of preservation. Sex and age-at-death were estimated through classical anthropological methods. Health status was inferred through the biological index of frailty (BIF) and paleopathological analysis was performed through macroscopic and radiographic investigations. The "bioarcheology of care" approach was applied to individuals who showed evidence of impairment and disability. RESULTS: The skeletal assemblage of St. Biagio was equally represented by males and females (50% males), with a higher percentage of adults (83.4%) than subadults (10.5%), and this is reflected in the high life expectancy at birth (40.3 years). No significant differences in health status emerged between age groups and sexes, with a generally high percentage of joint diseases, antemortem trauma, and infectious diseases. Evidence of care and compassion was found in some individuals with a high degree of impairment or disability, as in the case of probable Angelman syndrome. DISCUSSION: This study provided important insights into the biological and social aspects of an Early Modern population in Northern Italy, showing that people with functional and/or visible abnormalities were probably cared for in life and were presumably considered full members of the society.

Bioarchaeological and molecular evidence of tuberculosis in human skeletal remains from 18th-19th century orthodox cemeteries in Irkutsk, Eastern Siberia.

In this study, we tested the skeletal human remains from the 18th - early 19th century Orthodox cemeteries in Irkutsk, Eastern Siberia, for tuberculosis-associated morphological alterations and Mycobacterium tuberculosis DNA. The morphologically studied bone collection included 591 individuals of mainly Caucasian origin. The molecular methods (IS6110-PCR and spoligotyping) suggested that at least four individuals (out of 15 TB-suspected, DNA-tested) were positive for the presence of M. tuberculosis DNA. All of them were males (3 maturus, 1 maturus senilis). Two of them date back to the second and third quarters of the 18th century, another to the last quarter of the 18th century, and the last one to the second half of the 19th century. The combined molecular analysis cautiously suggested presence of different strains and at least some of them represented not the currently predominant in Siberia Beijing genotype (M. tuberculosis East-Asian lineage) but strains of European origin. In conclusion, this study presented bioarchaeological and molecular evidence of tuberculosis in human skeletal remains from 18th-19th century Orthodox cemeteries in Irkutsk, Eastern Siberia. The samples are not M. bovis and represent human M. tuberculosis sensu stricto. Their precise phylogenetic identity is elusive but evokes the European/Russian origin of at least some isolates.

Extraction Protocol for Parallel Analysis of Proteins and DNA from Ancient Teeth and Dental Calculus.

Dental calculus is becoming a crucial material in the study of past populations with increasing interest in its proteomic and genomic content. Here, we suggest further development of a protocol for analysis of ancient proteins and a combined approach for subsequent ancient DNA extraction. We tested the protocol on recent teeth, and the optimized protocol was applied to ancient tooth to limit the destruction of calculus as it is a precious and irreplaceable source of dietary, microbiological, and ecological information in the archeological context. Finally, the applicability of the protocol was demonstrated on samples of the ancient calculus.

The Late Iron Age in Switzerland: a review of anthropological, funerary, and isotopic studies.

The Iron Age in continental Europe is a period of profound cultural and biological importance with heterogeneous trends through space and time. Regional overviews are therefore useful for better understanding the main cultural and biological patterns characterizing this period across the European regions. For the area of modern Switzerland, a rich archeological and anthropological record represents the Late Iron Age. However, no review of the main anthropological and funerary patterns for this period is available to date. Here we assess the available demographic, paleopathological, funerary, and isotopic data for the Late Iron Age in the Swiss territory, and summarize the cultural and biological patterns emerging from the available literature. Finally, we highlight a series of research avenues for future studies.

Investigating mobility and pastoralism in Kerma-period communities upstream of the fourth cataract, Sudan.

OBJECTIVES: The Kingdom of Kush in today's northern Sudan and southern Egypt (ancient Nubia) is often depicted as a secondary state relative to ancient Egypt. More recent investigations have set aside Egyptocentric and western, colonialist perspectives of state development focused on control of land and agricultural surplus, examining Kushites through the lens of African-based models of mobile pastoralism in which power and authority were achieved through control of herds and alliance-building. Here, analyses of radiogenic strontium isotopes in human dental enamel are used to investigate diachronic shifts in mobility patterns linked to pastoralism and state development during the Kerma period (ca. 2500-1100 BCE). MATERIALS AND METHODS: From five cemetery sites around al Qinifab, Sudan, upstream of the capital at Kerma, we analyzed the strontium isotope ratios of 50 teeth from 27 individuals dating from the Early through Late Kerma phases. RESULTS: Individuals from the Early and Middle Kerma phases demonstrated considerable 87 Sr/86 Sr ratio variability (mean = 0.70835 ± 0.00109), with 50% falling outside the locally bioavailable strontium range. Conversely, most Classic (0.70756 ± 0.00043) and Late Kerma (0.70755 ± 0.00036) individuals exhibited ratios consistent with the local region. DISCUSSION: These changes indicate a potential transformation in subsistence strategies and social organization as early communities engaged in a more mobile lifestyle than later groups, suggesting a greater degree of pastoralism followed by declining mobility with Kushite state coalescence and a shift to agropastoralism. Because 87 Sr/86 Sr ratios from enamel reflect childhood geographic residence, these findings indicate that mobility likely involved extended family groups, and not just transhumant adults.

Adapting in the Arctic II: Upper limb diaphyseal robusticity and habitual activity in Late Holocene hunter-gatherers from Alaska.

OBJECTIVES: This study compares humeral diaphyseal robusticity and asymmetry between Late Holocene hunter-gatherers from Alaska with the goal of reconstructing habitual activity in relation to culture and environment. MATERIALS AND METHODS: Ancestral remains from four geographic regions of Alaska were divided into five site groups defined by subsistence strategies and technology: Aleutian Islands, Coastal Bay, Far North Coastal, Inland/Riverine, and Tikeraq. Mid-distal humeral diaphyseal robusticity was quantified using cross-sectional geometric properties standardized by estimated body mass and bone length. RESULTS: Humeral strength and bilateral asymmetry were greatest in Aleutian Island males, moderate in Far North Coastal and Tikeraq males, and reduced in Inland/Riverine males. Left-biased directional asymmetry and reduced humeral strength were found in Coastal Bay males. Aleutian Island males had relatively mediolaterally strengthened humeri compared with other groups. Aleutian Island females had elevated humeral strength, while humeral asymmetry among females was moderate and did not vary between groups. Humeri were relatively round among Aleutian Island and Tikeraq females and anteroposteriorly (A-P) strengthened among Coastal Bay, Far North Coastal, and Inland/Riverine females. CONCLUSIONS: These results suggest elevated humeral strength and asymmetry in males that engaged in rowing and unimanual projectile hunting, while reduced humeral strength and asymmetry may reflect bow-and-arrow or ensnarement technologies. Left-biased humeral asymmetry may be associated with net-fishing. Humeral strength and asymmetry are consistent with select instances of unimanual projectile hunting in females, while differences in humeral A-P/mediolateral strength may reflect variation in butchery and processing of prey versus rowing and throwing behaviors.

Application of natural sciences methodology in archaeological study of Iron Age burials in Latvia: pilot study.

Natural sciences provide several modern methodologies that could be successfully applied in archaeological studies. In this pilot study, archaeological human remains from two Iron Age cemeteries (7th-twelfth centuries AD), Lejasbiteni and Cunkani-Drengeri, which are located in different regions of Latvia, were studied. We applied ancient DNA (aDNA) and tooth enamel peptide analysis to determine the biological sex of the individuals. In addition, aDNA analysis was used to perform mtDNA haplogroup analysis. In most cases, the results of aDNA analysis regarding the biological sex of individuals coincided with the gender assigned based on grave orientation and grave goods. The results of sex determination using peptide analysis in all four individuals for whom data were available matched the possible gender. Of the 17 samples that had sufficient DNA for sequencing, seven samples had enough reads to perform mtDNA haplogroup analysis. The H2a2a, I4a1, H2a2a1, and H16c mtDNA haplogroups were identified in the individuals from the Lejasbiteni cemetery, while the T2b and K1a + 150 mtDNA haplogroups were identified in the individuals from the Cunkani-Drengeri cemetery. Overall, the obtained results demonstrated the feasibility of applying aDNA and tooth enamel peptide analysis for biological sex determination within archaeological studies. The availability of human aDNA data will be highly useful for investigating the demographic history and social structures in Iron Age Latvia.

Pathology or expected morphology? Investigating patterns of cortical porosity and trabecularization during infancy and early childhood.

Increased cortical porosity is associated with a heightened risk of skeletal fragility due to bone loss and structural decay in adults. However, few studies have examined the etiology of cortical porosity in infants and children. This study examines whether age-related changes in femoral growth and locomotor development influence femoral midshaft cortical porosity in a sample of 48 individuals (fetal to 3.99 years) from the 10th-13th century cemetery of St. Étienne de Toulouse, France. Histological sections were prepared and imaged using light microscopy. Midshaft geometric variables such as total area, cortical area, and pore area were calculated using BoneJ. Increased porosity and cortical trabecularization were found to be significantly associated with age, being almost exclusively present in individuals aged 0.5-1.99 years. At approximately 6 months of age infants typically begin engaging in regular femoral loading and experience an acceleration in growth. The observed increase in midshaft porosity and trabecularization, therefore, likely results from the reorganization and redistribution of cortical bone, stimulated by increased growth velocity and the onset of weight-bearing activities. The reduction in cortical porosity and trabecularization in individuals aged 2.0-3.99 years indicates that children are approaching some sort of homeostasis as growth velocity slows and their femora adapt to consistent loading. Understanding what expected skeletal development looks like is necessary when conducting bioarcheological studies and this study provides evidence for a pattern of transient midshaft porosity during infancy and early childhood.

A Case of Acute Plastic Deformation of the Forearm in a Medieval Hispano-Mudejar Skeleton (13-14th Centuries AD).

INTRODUCTION: Acute plastic deformation refers to a traumatic bending or bowing without a detectable cortical defect. CASE PRESENTATION AND DISCUSSION: We describe a rare case from an individual that was exhumed from the Hispano-Mudejar necropolis in Uceda (Guadalajara, Spain) dated between the 13th and 14th centuries AD. The case corresponds to an adult woman, with a bowing involvement of the left ulna and radius. After making the differential diagnosis with various pathologies likely to present with this alteration, we reached the diagnosis of acute plastic deformation of the forearm through external and radiological examination and comparison with the healthy contralateral forearm. CONCLUSIONS: Acute plastic deformation is a rare traumatic injury, not described until the last century and only rarely described in palaeopathological contexts. We contribute a new case, the first being sufficiently documented, contributing to the knowledge and diagnosis of this type of trauma in the ancient bone, while deepening the knowledge of the living conditions of the medieval Mudejar population of Uceda.

Origin, Evolution, and Research Development of Donkeys.

Lack of archaeological and whole-genome diversity data has restricted current knowledge of the evolutionary history of donkeys. With the advancement of science and technology, the discovery of archaeological evidence, the development of molecular genetics, and the improvement of whole-genome sequencing technology, the in-depth understanding of the origin and domestication of donkeys has been enhanced, however. Given the lack of systematic research, the present study carefully screened and collected multiple academic papers and books, journals, and literature on donkeys over the past 15 years. The origin and domestication of donkeys are reviewed in this paper from the aspects of basic information, cultural origin, bioarcheology, mitochondrial and chromosomal microsatellite sequences, and whole-genome sequence comparison. It also highlights and reviews genome assembly technology, by assembling the genome of an individual organism and comparing it with related sample genomes, which can be used to produce more accurate results through big data statistics, analysis, and computational correlation models. Background: The donkey industry in the world and especially in China is developing rapidly, and donkey farming is transforming gradually from the family farming model to large-scale, intensive, and integrated industrial operations, which could ensure the stability of product quality and quantity. However, theoretical research on donkey breeding and its technical development lags far behind that of other livestock, thereby limiting its industrial development. This review provides holistic information for the donkey industry and researchers, that could promote theoretical research, genomic selection (GS), and reproductive management of the donkey population.

Review of United States laws pertaining to the recovery and analysis of human skeletal remains.

Human skeletal remains (HSR) are routinely excavated from archeological contexts and analyzed by experts in human osteology. Contrarily, HSR in medicolegal contexts are usually recovered by law enforcement officers and examined by pathologists with limited osteological training. To examine legal requirements for expertise, we reviewed laws in the United States regarding the recovery and analysis of HSR from archeological sites, unmarked graves, and medicolegal contexts. Of the 50 states, 19 (38%) have laws stating that an anthropologist with osteological training should be involved in the recovery or analysis of HSR from an archeological context. Fifteen of those 16 states have laws requiring a minimum level of education to be a qualified skeletal analyst. In contrast, only one state, Texas, requires an anthropologist who handles forensic cases to have a doctoral degree. Including Texas, only eight states (16%) have laws that encourage but do not mandate consultation with a forensic anthropologist for medicolegal skeletal cases. Louisiana and Washington have state-funded laboratories, expert forensic anthropologists, and effective protocols for handling forensic cases. Due process and human rights concerns at stake in criminal cases require that those recovering and analyzing modern HSR have an equal or higher level of expertise than those working with archeological remains. Yet, legislators assume that law enforcement and pathologists are adequately trained. Because court standards demand expert testimony based on accepted methodologies and standard levels of competency, forensic anthropologists have a professional responsibility to engage with lawmakers to draft legislation to ensure proper handling of all skeletal cases.

DXAGE 2.0 - adult age at death estimation using bone loss in the proximal femur and the second metacarpal.

The accurate age at death assessment of unidentified adult skeletal individuals is a critical research task in forensic anthropology, being a key feature for the determination of biological profiles of individual skeletal remains. We have previously shown that the age-related decrease of bone mineral density (BMD) in the proximal femur could be used to assess age at death in women (Navega et al., J Forensic Sci 63:497-503, 2018). The present study aims to generate models for age estimation in both sexes through bone densitometry of the femur and radiogrammetry of the second metacarpal. The training sample comprised 224 adults (120 females, 104 males) from the "Coimbra Identified Skeletal Collection," and different models were generated through least squares regression and general regression neural networks (GRNN). The models were operationalized in a user-friendly online interface at https://osteomics.com/DXAGE2/ . The mean absolute difference between the known and estimated age at death ranges from 9.39 to 13.18 years among women and from 10.33 to 15.76 among men with the least squares regression models. For the GRNN models, the mean absolute difference between documented and projected age ranges from 8.44 to 12.58 years in women and from 10.56 to 16.18 years in men. DXAGE 2.0 enables age estimation in incomplete and/or fragmentary skeletal remains, using alternative skeletal regions, with reliable results.

A Healthier Smile in the Past? Dental Caries and Diet in Early Neolithic Farming Communities from Central Germany.

Dental health is closely linked to an individual's health and diet. This bioarcheological study presents dental caries and stable isotope data obtained from prehistoric individuals (n = 101) from three Early Neolithic sites (c. 5500-4800 BCE) in central Germany. Dental caries and ante-mortem tooth loss (AMTL) were recorded and related to life history traits such as biological sex and age at death. Further, we correlate evidence on caries to carbon and nitrogen isotope data obtained from 83 individuals to assess the relationship between diet and caries. In 68.3% of the adults, carious lesions were present, with 10.3% of teeth affected. If AMTL is considered, the values increase by about 3%. The prevalence of subadults (18.4%) was significantly lower, with 1.8% carious teeth. The number of carious teeth correlated significantly with age but not sex. The isotopic data indicated an omnivorous terrestrial diet composed of domestic plants and animal derived protein but did not correlate with the prevalence of carious lesions. The combined evidence from caries and isotope analysis suggests a prevalence of starchy foods such as cereals in the diet of these early farmers, which aligns well with observations from other Early Neolithic sites but contrasts to Late Neolithic and Early Bronze Age populations in Germany.

Historical bioarcheological aspects of diffuse idiopathic skeletal hyperostosis in humans.

OBJECTIVE: To identify diffuse idiopathic skeletal hyperostosis (DISH) in the human bioarcheological record to seek out temporal, geographic and dietary information to enhance better understanding of this common condition. MATERIALS AND METHODS: A review of available literature was conducted. RESULTS: DISH has been identified in hominin populations over millions of years, including several different human species. The distribution of DISH in ancient populations is diverse, both temporally and geographically. Where available, dietary intake of subjects with DISH, in contrast to those without DISH, suggests that metabolic factors associate with DISH. CONCLUSION: DISH is a ubiquitous human disorder over the ages. Metabolic factors appear important in ancient populations of those with DISH.

Prevalence of Dental Caries in Past European Populations: A Systematic Review.

OBJECTIVE: Within the fields of anthropology and osteoarcheology, human teeth have long been studied to understand the diet, habits, and diseases of past civilizations. However, no complete review has been published to collect and analyze the extensive available data on caries prevalence in European man (Homo sapiens) over time. METHOD: In this current study, the two databases, Scopus and Art, Design, and Architecture Collection, were searched using predefined search terms. The literature was systematically reviewed and assessed by two of the authors. RESULTS: The findings include a significant nonlinear correlation with increasing caries prevalence in European populations from 9000 BC to 1850 AD, for both the number of carious teeth and the number of affected individuals. CONCLUSION: Despite the well-established collective belief that caries rates fluctuate between different locations and time and the general view that caries rates have increased from prehistoric times and onwards, this is to our knowledge the first time this relationship has been proven based on published data.

A possible case of paralysis in early modern Vilnius and the implications for social care.

OBJECTIVE: To present a case of possible paralysis from early modern Vilnius and to discuss the potential level of care that was provided in the society of that time. MATERIALS: A partially disturbed skeleton of a young female from a 16th-17th century Orthodox Christian cemetery. METHODS: Macroscopic, osteometric and X-ray examinations coupled with a literature review aimed at providing a differential diagnosis. RESULTS: The skeletal remains showed signs of disuse atrophy most probably due to a neurological disorder acquired in the woman's late teens. Differentials suggest that the observed limb atrophy was most likely a consequence of poliomyelitis. CONCLUSIONS: The case of a young female with paralysis presented in this paper could serve as an example of care provided by her household. SIGNIFICANCE: This study substantially contributes to further understanding of the nature and quality of care provided to disabled individuals in their households even in the absence of written sources. LIMITATIONS: There is a degree of diagnostic ambiguity due to the application of routine clinical criteria to paleopathological cases. SUGGESTIONS FOR FURTHER RESEARCH: The article makes several recommendations for future research, e.g., systematic investigation of possible cases of bone atrophy in a broader sociocultural context, as well as searching for evidence of gastrointestinal infections, especially poliomyelitis, supplemented by the application of biomolecular technologies.

Isolation and Characterization of Live Yeast Cells from Ancient Clay Vessels.

Ancient fermented food has been studied mainly based on residue analysis and recipes and reconstruction attempts were performed using modern domesticated yeast. Furthermore, microorganisms which participated in fermentation were studied using ancient-DNA techniques. In a recent paper, we presented a novel approach based on the hypothesis that enriched yeast populations in fermented beverages could have become the dominant species in storage vessels and their descendants could be isolated and studied today. Here we present a pipeline for isolation of yeast from clay vessels uncovered in archeological sites and transferred to the microbiology lab where they can be isolated and characterized. This method opens new avenues for experimental archeology and enables attempts to recreate ancient food and beverages using the original microorganisms.