Entheseal variation and locomotor behavior during growth.

Entheses are acknowledged as skeletal markers capable of revealing several biological and behavioral aspects of past individuals and populations. However, entheseal changes (ECs) of juvenile individuals have not yet been studied with a systematic approach. This contribution aims at investigating the morphological changes occurring at the femoral insertion of the gluteus maximus and tibial origin of the soleus muscles to highlight a potential link between the morphological features of those entheses and skeletal maturity in relation to sex, age, and locomotor developmental patterns. The sample consisted of 119 skeletons (age-at-death: 0-30 years) belonging to the Documented Human Skeletal Collection of the Certosa Cemetery (Bologna, Italy). The entheseal variation during the last stages of skeletal maturation in young adults was assessed using existing recording standards. A recording protocol for each enthesis was developed for immature individuals to subdivide the morphological variability into discrete categories. Univariate, bivariate, and multivariate statistical analyses were performed to investigate the variation of entheseal morphologies and measurements in relation to bone metrics, degree of epiphyseal closure, sex, age, and locomotor developmental patterns. A statistically significant relationship was observed between ECs morphological patterns and age for both entheses, while sexual differences were negligible. A relationship between ECs morphological pattern and locomotor milestones emerged only for the gluteus maximus. Even though further testing is needed on other documented skeletal collections, our protocol could be usefully applied in forensic and archaeological fields and serving as important reference for evolutionary investigations.

Ancient genomes reveal a high diversity of Mycobacterium leprae in medieval Europe.

Studying ancient DNA allows us to retrace the evolutionary history of human pathogens, such as Mycobacterium leprae, the main causative agent of leprosy. Leprosy is one of the oldest recorded and most stigmatizing diseases in human history. The disease was prevalent in Europe until the 16th century and is still endemic in many countries with over 200,000 new cases reported annually. Previous worldwide studies on modern and European medieval M. leprae genomes revealed that they cluster into several distinct branches of which two were present in medieval Northwestern Europe. In this study, we analyzed 10 new medieval M. leprae genomes including the so far oldest M. leprae genome from one of the earliest known cases of leprosy in the United Kingdom-a skeleton from the Great Chesterford cemetery with a calibrated age of 415-545 C.E. This dataset provides a genetic time transect of M. leprae diversity in Europe over the past 1500 years. We find M. leprae strains from four distinct branches to be present in the Early Medieval Period, and strains from three different branches were detected within a single cemetery from the High Medieval Period. Altogether these findings suggest a higher genetic diversity of M. leprae strains in medieval Europe at various time points than previously assumed. The resulting more complex picture of the past phylogeography of leprosy in Europe impacts current phylogeographical models of M. leprae dissemination. It suggests alternative models for the past spread of leprosy such as a wide spread prevalence of strains from different branches in Eurasia already in Antiquity or maybe even an origin in Western Eurasia. Furthermore, these results highlight how studying ancient M. leprae strains improves understanding the history of leprosy worldwide.

The study of the lower limb entheses in the Neanderthal sample from El Sidrón (Asturias, Spain): How much musculoskeletal variability did Neanderthals accumulate?

Entheses have rarely been systematically studied in the field of human evolution. However, the investigation of their morphological variability (e.g., robusticity) could provide new insight into their evolutionary significance in the European Neanderthal populations. The aim of this work is to study the entheses and joint features of the lower limbs of El Sidrón Neanderthals (Spain; 49 ka), using standardized scoring methods developed on modern samples. Paleobiology, growth, and development of both juveniles and adults from El Sidrón are studied and compared with those of Krapina Neanderthals (Croatia, 130 ka) and extant humans. The morphological patterns of the gluteus maximus and vastus intermedius entheses in El Sidrón, Krapina, and modern humans differ from one another. Both Neanderthal groups show a definite enthesis design for the gluteus maximus, with little intrapopulation variability with respect to modern humans, who are characterized by a wider range of morphological variability. The gluteus maximus enthesis in the El Sidrón sample shows the osseous features of fibrous entheses, as in modern humans, whereas the Krapina sample shows the aspects of fibrocartilaginous ones. The morphology and anatomical pattern of this enthesis has already been established during growth in all three human groups. One of two and three of five adult femurs from El Sidrón and from Krapina, respectively, show the imprint of the vastus intermedius, which is absent among juveniles from those Neanderthal samples and in modern samples. The scant intrapopulation and the high interpopulation variability in the two Neanderthal samples is likely due to a long-term history of small, isolated populations with high levels of inbreeding, who also lived in different ecological conditions. The comparison of different anatomical entheseal patterns (fibrous vs. fibrocartilaginous) in the Neanderthals and modern humans provides additional elements in the discussion of their functional and genetic origin.

Estimation of adult age-at-death from entheseal robusticity: A test using an identified Italian skeletal collection.

OBJECTIVES: The present study tests the effectiveness of entheseal robusticity (ER) as a potential predictor of adult age-at-death by applying multiple regression models to a large contemporary identified skeletal sample. MATERIALS AND METHODS: ER was recorded for 23 bilateral postcranial entheses on 481 adult individuals (271 females and 210 males) from the Frassetto identified skeletal collection of Sassari (Italy), following the method of Mariotti et al. 2007. ER scores were used as predictors in multiple regression analyses with age as the dependent variable; the performance of the resulting models was tested through standard error of estimate, the correlation coefficient between predicted and documented age, and the percentage of correctly classified individuals. RESULTS: ER show a higher correlation with age in females, resulting in more accurate estimates when compared with those obtained for males. Age-at-death is overestimated for male individuals under 45 years old and underestimated for older individuals. Regression models including a reduced set of ER sites result in better estimates compared with those using the complete set of variables. DISCUSSION: Results suggest the potential usefulness of ER for the estimation of age, especially for female skeletal remains. If used with caution, ER may be a valid complement to existing methods for the reconstruction of the biological profile of skeletal remains of archaeological and forensic interest.

Variations in epiphyseal fusion and persistence of the epiphyseal line in the appendicular skeleton of two identified modern (19th-20th c.) adult Portuguese and Italian samples.

OBJECTIVES: The aim of this work is to study age, sex, and population variations in epiphyseal fusion and persistence of the epiphyseal line in the appendicular skeleton of two identified modern (19th-20th c.) adult skeletal samples, using a specifically designed macroscopic scoring method. The use of epiphyseal closure and persistence of the epiphyseal line as an adult-age marker is also discussed. MATERIALS AND METHODS: This study examined 981 adult skeletons of both sexes from two identified modern (20th c.) skeletal samples from the Sassari Collection (Museum of Anthropology, University of Bologna, Italy) and the Colecção de Esqueletos Identificados (Museum of Anthropology, University of Coimbra, Portugal). Our scoring method considers a five-degree scale, from nonfusion (Degree 0) to complete fusion (Degree 4). In addition, the persistence of the epiphyseal line, a feature that is not commonly collected during routine anthropological analyses, is taken into account here as Degree 3. RESULTS: Intra- and interobserver errors of 1.2% and 5.2%, respectively, were found, suggesting a good reproducibility of this scoring method. Some sites show variable degrees of epiphyseal fusion still in adult skeletons (e.g., secondary center of ossification of the clavicle, iliac crest, ischial tuberosity, distal epiphysis of the radius and ulna). CONCLUSIONS: Population differences have been observed, showing a delay in the complete epiphyseal closure for the Sassari sample compared to the Coimbra sample. Degree 3 seems to be a good adult-age indicator for individuals less than 35-year-old.

Tooth fractures in the Krapina Neandertals.

Dental fractures can be produced during life or post-mortem. Ante-mortem chipping may be indicative of different uses of the dentition in masticatory and non-masticatory activities related to variable diets and behaviors. The Krapina collection (Croatia, 130,000 years BP), thanks to the large number of teeth (293 teeth and tooth fragments) within it, offers an excellent sample to investigate dental fractures systematically. Recorded were the distribution, position and severity of the ante-mortem fractures according to standardized methods. High frequencies of teeth with chipping in both Krapina adults and subadults suggest that the permanent and deciduous dentition were heavily subjected to mechanical stress. This is particularly evident when the frequencies of chipping are compared with those in modern humans (Upper Paleolithic and historic samples) that we analysed using the same methods. The distribution of chipping in the Krapina sample (anterior teeth are more affected) and its position (labial) suggest a systematic use of the anterior teeth for non-masticatory tasks.

A muscular imprint on the anterolateral surface of the proximal femurs of the Krapina Neandertal collection.

OBJECTIVES: The purpose of this study is to report and interpret a feature on the anterolateral surface of the proximal femurs of the Krapina hominid collection that we briefly described in 2006 (Periodicum Biologorum, 108, 319-329). MATERIALS AND METHODS: We recorded the presence or absence of the feature in all the proximal femurs of the Krapina collection (six specimens recordable) and in 622 modern human adult femurs. RESULTS: The feature consists in a series of crests delimitating three raised or depressed areas. This feature has been found in three out of four adult Neandertal femurs observable. The two observable subadult Neandertal femurs do not show this character. None of the modern femurs displayed the feature. CONCLUSION: We interpret this feature as a muscular imprint, probably representing the m. vastus intermedius origin and discuss a possible interpretation. We did not find any other references for such imprint in the existing literature regarding the Neandertal femurs.

Variation of skeletal degenerative joint disease features in an identified Italian modern skeletal collection.

OBJECTIVES: The objective of this study was to investigate the relationship of degenerative joint disease (DJD) features (marginal lipping, porosity, surface osteophytes, eburnation, and loss of joint morphology) with sex, age, and occupation. MATERIALS AND METHODS: We devised a recording method for scoring the single features in the shoulder, elbow, wrist, hip, knee, and ankle of an Italian skeletal sample of 332 individuals (males = 161, females = 171) of known sex, age, and occupation from the Frassetto collection (Sassari, Sardinia, 19th-20th century). RESULTS: Our data revealed that: (1) the frequency of all features increases with age; (2) porosity is the first feature to appear, being in general the most frequent feature in young adults; (3) sex differences are especially found in the upper limbs of mature and elderly adults; (4) marginal lipping and surface osteophytes show a lesser age correlation and, in general, higher frequencies in the left lower limb; (5) no differences were found among different occupations. DISCUSSION: We explain these results in light of different metabolic and adaptive processes in relation to age and sex. Static (weight-bearing) rather than dynamic mechanical stress could play a pivotal role in the distribution of the features observed in the lower limbs. The considerable increase in marginal lipping and surface osteophytes in elderly adults may be an adaptation for sustaining mechanical loading, balancing the loss of bone tissue which is characteristic of ageing processes (osteopenia, osteoporosis). Am J Phys Anthropol, 2016. © 2016 Wiley Periodicals, Inc. Am J Phys Anthropol 160:683-693, 2016. © 2016 Wiley Periodicals, Inc.

Skeletal evidence of tuberculosis in a modern identified human skeletal collection (Certosa cemetery, Bologna, Italy).

The diagnosis of tuberculosis (TB) in osteoarcheological series relies on the identification of osseous lesions caused by the disease. The study of identified skeletal collections provides the opportunity to investigate the distribution of skeletal lesions in relation to this disease. The aim of this study was to examine the skeletal evidence for TB in late adolescent and adult individuals from the identified human collection of the Certosa cemetery of Bologna (Italy, 19th-20th c.). The sample group consists of 244 individuals (138 males, 106 females) ranging from 17 to 88 years of age. The sample was divided into three groups on the basis of the recorded cause of death: TB (N = 64), pulmonary non-TB (N = 29), and other diseases (N = 151). Skeletal lesions reported to be related to TB were analyzed. The vertebral lesions were classified into three types: enlarged foramina (EnF, vascular foramina with diameter of 3-5 mm), erosions (ER), and other foramina (OtF, cavities of various shapes > 3 mm). A CT scan analysis was also performed on vertebral bodies. Some lesions were seldom present in our sample (e.g., tuberculous arthritis). OtF (23.7%) and subperiosteal new bone formation on ribs (54.2%) are significantly more frequent in the TB group with respect to the other groups. The CT scan analysis showed that the vertebrae of individuals who have died of TB may have internal cavities in the absence of external lesions. These traits represent useful elements in the paleopathological diagnosis of TB.

Histotaphonomic analysis of bone bioerosion reveals a regional framework of diverse deathways in the Neolithic of Southeast Italy.

The wide diversity of Neolithic funerary practices is increasingly recognised. In Southeast Italy, recent studies have drawn attention to the co-existence of multiple ways of treating the dead within single sites and across the region. In this study, we address how such diverse deathways form a regional framework of ritual practice through histotaphonomic analysis of bone bioerosion. Samples were obtained from articulated, semi-articulated and disarticulated remains from four sites in Apulia which each presented different modes of treatment and disposal of the dead. Bone thin sections were analysed by light microscopy to characterise microstructural preservation through features including bacterial bioerosion, staining, inclusions, and Wedl tunnelling. We investigate the early post-mortem histories of individuals whose remains ended up in various states of dis/articulation and diverse depositional contexts. Disarticulated remains frequently displayed arrested or extensive bacterial bioerosion, which was also found in articulated and semi-articulated skeletons. Additionally, remains deposited in similar contexts, as well as articulated and disarticulated remains deposited together in the same context, often showed different histotaphonomic characteristics, suggesting diverse early post-mortem trajectories. As a result, we argue that Neolithic deathways in southeastern Italy incorporated a high level of diversity in the early post-mortem treatment of the body. A framework for funerary practices emerges, whereby disarticulated remains probably originated from bodies which had been buried previously and subjected to varying extents of shelter, exposure to invertebrates, and duration of burial. However, we acknowledge the ongoing research into the origins of bacterial bioerosion and the problem of equifinality, which leaves open the possibility for further scenarios of early post-mortem treatment.

Variation of the anterior aspect of the femoral head-neck junction in a modern human identified skeletal collection.

The effectiveness of the so-called skeletal markers of activity as functional indicators is widely debated. Among them, certain morphological features of the anterior aspect of the femoral head-neck junction (Poirier's facet, cervical fossa of Allen, etc.) have been considered in relation to some behaviors and specific activities (e.g., squatting, horseback riding, etc.). However, disagreement on terminology and descriptions, the absence of standardized scoring methods and poor knowledge of the variability and distribution of these features make it difficult to interpret their meaning. The aim of this study is to analyze the variability of the anterior aspect of the femoral neck through a new scoring method taking into account three main traits: Poirier's facet, plaque, and cribra (including the Allen's fossa). This method has been applied to a sample of 225 adult individuals of both sexes coming from an identified modern skeletal collection, achieving low intraobserver and interobserver error values. The results highlight some significant trends: plaque, almost always bilateral, appears to be a normal condition of the femur, being present in approximately 90% of the individuals. Cribra is more frequent in females and decreases with age. Poirier's facet shows a very low frequency. This method allows the representation of both the anatomical diversity of the region already described in literature and part of the variability never considered before. Our results suggest caution in considering these features as markers related to specific activities.

Osteobiography of a 19th century elderly woman with pertrochanteric fracture and osteoporosis: a multidisciplinary approach.

In this paper the osteobiography of an elderly woman recovered from a cemetery tomb where she was buried in 1850, affected by hip fracture and osteoporosis, is described. The overall anthropological characteristics of the individual have been investigated. Macroscopic, radiographic, tomographic, microscopic, and chemical and structural examinations have been performed to give a detailed account of the condition of the skeleton. A non-union pertrochanteric fracture not surgically treated and probably due to senile osteoporosis was diagnosed. The consequences of the fracture to the bones show that this individual likely survived several years following the injury. The osseous features we describe (remodelled bone at the fracture site, asymmetry of entheseal changes likely related to the particular walking pattern of the individual) may be useful in personal identification of skeletons of legal interest. Regarding the recognition of osteoporosis in unearthed skeletons, our study underlines that the cortical thickness, microscopic features, degree of crystallinity and Ca/P ratio represent more useful elements than the mean bone density, mineral/matrix ratio and mineral maturity, which are more sensitive to diagenetic changes that affect the mineral phase post-mortem.

On the traces of lost identities: chronological, anthropological and taphonomic analyses of the Late Neolithic/Early Eneolithic fragmented and commingled human remains from the Farneto rock shelter (Bologna, northern Italy).

The present study examines the prehistoric human skeletal remains retrieved starting from the 1920s in the deposit of the Farneto rock shelter, situated in the area of the 'Parco dei Gessi Bolognesi e Calanchi dell'Abbadessa' (San Lazzaro di Savena, Bologna, northern Italy). An exact dating and a reliable interpretation of the assemblage had not been reached so far because of the lack of contextual data useful for dating purposes, the inaccurate recovery procedures of the remains and their state of preservation. In fact, the skeletal remains from the Farneto rock shelter are highly fragmented and commingled, whereas reliable information about their original position and their recovery procedures are not available. Despite these difficulties, radiocarbon analyses allowed the precise dating of the remains to a final phase of the Neolithic and an early phase of the Eneolithic period in Emilia Romagna (northern Italy). The study of the assemblage enabled to clarify the use of the context for funerary purposes. Moreover, the anthropological and taphonomic analyses of the skeletal remains shed light on the biological profile of the individuals and on some events that occurred after their death. In particular, the analysis of perimortem lesions highlighted the existence of intentional interventions related to corpse treatment, referable to dismembering/disarticulation and scarnification, i.e. cleaning of bones from soft tissues. Finally, the comparison with other Italian and European Neo/Eneolithic funerary contexts enabled a better understanding of these complex ritual practices. Supplementary Information: The online version contains supplementary material available at 10.1007/s12520-023-01727-2.

The effect of age, sex, and physical activity on entheseal morphology in a contemporary Italian skeletal collection.

Entheseal changes are traditionally included in a large array of skeletal features commonly referred to as "skeletal markers of activity." However, medical studies and recent anthropological analyses of identified skeletal series suggest a complex combination of physiological and biomechanical factors underlying the variability of such "markers." The aim of this study is to examine the relationship between age, sex, physical activity, and entheseal variability. To this end, 23 postcranial entheses are examined in a large (N = 484) Italian contemporary skeletal series using standardized scoring methods. The sample comprises subjects of known age, sex and, mostly, occupation. Results show a strong relationship between age and entheseal changes. Differences between sexes are also highlighted, while the effects of physical activity appear moderate. Altogether, our study indicates that entheseal morphology primarily reflects the age of an individual, while correlation with lifetime activity remains ambiguous.

Ancient oral microbiomes support gradual Neolithic dietary shifts towards agriculture.

The human microbiome has recently become a valuable source of information about host life and health. To date little is known about how it may have evolved during key phases along our history, such as the Neolithic transition towards agriculture. Here, we shed light on the evolution experienced by the oral microbiome during this transition, comparing Palaeolithic hunter-gatherers with Neolithic and Copper Age farmers that populated a same restricted area in Italy. We integrate the analysis of 76 dental calculus oral microbiomes with the dietary information derived from the identification of embedded plant remains. We detect a stronger deviation from the hunter-gatherer microbiome composition in the last part of the Neolithic, while to a lesser extent in the early phases of the transition. Our findings demonstrate that the introduction of agriculture affected host microbiome, supporting the hypothesis of a gradual transition within the investigated populations.

Hyperostosis frontalis interna (HFI) and castration: the case of the famous singer Farinelli (1705-1782).

The famous castrato singer Farinelli (1705-1782) was exhumed by our research group in July 2006 for the purpose of gaining some insight into his biological profile through a study of his skeletal remains. Farinelli was castrated before puberty to preserve the treble pitch of the boy's voice into adult life. His powerful and sweet voice became legendary. In spite of its bad preservation state, the skeleton displayed some interesting characteristics that are probably related to the effects of castration, including long limb-bones, persistence of epiphyseal lines and osteoporosis. In particular, the frontal bone was affected by severe hyperostosis frontalis interna (HFI). This condition consists in a symmetrical thickening of the inner table of the bone. The epidemiology of HFI shows that it is relatively common in postmenopausal women but very rare in men. Men affected by this pathology suffer from diseases, syndromes or treatments causing androgen deficiency. In the case of Farinelli, castration was probably responsible for the onset and development of this lesion.

Lower limb entheseal morphology in the Neandertal Krapina population (Croatia, 130,000 BP).

Although the Neandertal locomotor system has been shown to differ from Homo sapiens, characteristics of Neandertal entheses, the skeletal attachments for muscles, tendons, ligaments and joint capsules, have never been specifically investigated. Here, we analyse lower limb entheses of the Krapina Neandertal bones (Croatia, 130,000 BP) with the aim of determining how they compare with modern humans, using a standard developed by our research group for describing modern human entheseal variability. The entheses examined are those of the gluteus maximus, iliopsoas and vastus medialis on the femur, the quadriceps tendon on the patella, and soleus on the tibia. For the entheses showing a different morphological pattern from H. sapiens, we discuss the possibility of recognising genetic versus environmental causes. Our results indicate that only the gluteus maximus enthesis (the gluteal tuberosity), falls out of the modern human range of variation. It displays morphological features that could imply histological differences from modern humans, in particular the presence of fibrocartilage. In both H. sapiens and the Krapina Neandertals, the morphological pattern of this enthesis is the same in adult and immature femurs. These results can be interpreted in light of genetic differences between the two hominins. The possibility of functional adaptations to higher levels of mechanical load during life in the Neandertals seems less likely. The particular morphology and large dimensions of the Krapina enthesis, and perhaps its fibrocartilaginous nature, could have been selected for in association with other pelvic and lower limb characteristics, even if genetic drift cannot be ruled out.

Preliminary study: voluntary food intake in dogs during tryptophan supplementation.

Tryptophan, a precursor of important molecules such as serotonin, melatonin and niacin, is an essential amino acid for dogs. In pigs, tryptophan supplementation has been shown to induce a significant increase in food intake. The aim of the present study was to assess whether long-term tryptophan supplementation increases voluntary food intake in dogs and to observe whether this was accompanied by a change in serum ghrelin. In the present study, sixteen adult Beagle dogs were used, with four male and four female dogs fed diets supplemented with tryptophan (1 g/dog per d) during 81 d (Trp) and four male and four female dogs that were not supplemented (control). A voluntary food intake test was performed during 5 d following the supplementation period. The Trp group tended to show a higher food intake during the voluntary food intake test (58.0 (SE 5.37) v. 77.5 (SE 3.65) g/kg metabolic weight per d; P = 0.074). No significant differences were found for serum ghrelin concentrations.

Unveiling an odd fate after death: The isolated Eneolithic cranium discovered in the Marcel Loubens Cave (Bologna, Northern Italy).

An isolated human cranium, dated to the early Eneolithic period, was discovered in 2015 at the top of a vertical shaft in the natural Marcel Loubens gypsum Cave (Bologna area, northern Italy). No other anthropological or archaeological remains were found inside the cave. In other caves of the same area anthropic and funerary use are attested from prehistory to more recent periods. We focused on investigating the circumstances surrounding the death of this individual, since the cranium shows signs of some lesions that appear to be the results of a perimortem manipulation probably carried out to remove soft tissues. Anthropological analyses revealed that the cranium belonged to a young woman. We analysed the taphonomic features and geological context to understand how and why the cranium ended up (accidentally or intentionally) in the cave. The analyses of both the sediments accumulated inside the cranium and the incrustations and pigmentation covering its outer surface suggested that it fell into the cave, drawn by a flow of water and mud, likely from the edges of a doline. The accidental nature of the event is also seemingly confirmed by some post-mortem lesions on the cranium. The comparison with other Eneolithic archaeological sites in northern Italy made it possible to interpret the find as likely being from a funerary or ritual context, in which corpse dismemberment (in particular the displacement of crania) was practiced.

Funerary practices of the Iberomaurusian population of Taforalt (Tafoughalt, Morocco, 11-12,000 BP): the case of Grave XII.

The Iberomaurusian necropolis of Taforalt (Morocco, 11-12000 BP), excavated by Roche in the 1950s, contains 28 multiple graves. The funerary practices of the Taforalt population have been the focus of a previous work (Mariotti et al., 2009). In the absence of the excavation records of the necropolis, these funerary practices were investigated through the analysis of the contents of each grave and the distribution of intentionally modified specimens (ochre-dyeing, cut marks). Previous research has drawn particular attention to Grave XII (containing three male adults and two juveniles), where many intentionally modified specimens were identified. The present study focused specifically on the human remains recovered from Grave XII. Analysis of these remains has provided evidence of interventions, such as dismemberment and defleshing of the cadaver, and the use of ochre to colour the bones. Furthermore, the presence of lesions on two skulls suggests the possibility of intentional killing and cannibalism among the Taforalt population. This study further supports our previous impression of the complex and diversified funerary practices, characterising the social life of the Iberomaurusian population of Taforalt.