Strontium Isotope Signals in Cremated Petrous Portions as Indicator for Childhood Origin

Dental enamel is currently of high informative value in studies concerning childhood origin and human mobility because the strontium isotope ratio in human dental enamel is indicative of geographical origin. However, many prehistoric burials involve cremation and although strontium retains its original biological isotopic composition, even when exposed to very high temperatures, intact dental enamel is rarely preserved in cremated or burned human remains. When preserved, fragments of dental enamel may be difficult to recognize and identify. Finding a substitute material for strontium isotope analysis of burned human remains, reflecting childhood values, is hence of high priority. This is the first study comparing strontium isotope ratios from cremated and non-cremated petrous portions with enamel as indicator for childhood origin. We show how strontium isotope ratios in the otic capsule of the petrous portion of the inner ear are highly correlated with strontium isotope ratios in dental enamel from the same individual, whether inhumed or cremated. This implies that strontium isotope ratios in the petrous bone, which practically always survives cremation, are indicative of childhood origin for human skeletal remains. Hence, the petrous bone is ideal as a substitute material for strontium isotope analysis of burned human remains.     

Early African Diaspora in colonial Campeche, Mexico: strontium isotopic evidence

Construction activities around Campeche's central park led to the discovery of an early colonial church and an associated burial ground, in use from the mid-16th century AD to the late 17th century. Remains of some individuals revealed dental mutilations characteristic of West Africa. Analyses of strontium isotopes of dental enamel from these individuals yielded unusually high (87)Sr/(86)Sr ratios, inconsistent with an origin in Mesoamerica, but consistent with an origin in West Africa in terrain underlain by the West Africa Craton, perhaps near the port of Elmina, a principal source of slaves for the New World during the 16th century. These individuals likely represent some of the earliest representatives of the African Diaspora in the Americas.     

Multi-isotopic analysis reveals individual mobility and diet at the Early Iron Age monumental tumulus of Magdalenenberg, Germany

For the Early Iron Age western Hallstatt culture, which includes the site of Magdalenenberg in southwest Germany, it has been proposed that people were mobile and maintained far reaching social and trading networks throughout Europe. We tested this hypothesis by analyzing multiple isotopes (strontium, oxygen, sulfur, carbon, and nitrogen) of the preserved skeletons from the Magdalenenberg elite cemetery to determine diets and to look for evidence of mobility. The analysis of carbon, nitrogen, and sulfur isotope ratios in collagen of humans (n = 50) and associated domestic fauna (n = 10) indicates a terrestrial-based diet. There was a heterogeneous range of isotope values in both strontium (0.70725 to 0.71923, n = 76) and oxygen (13.4‰ to 18.5‰, n = 78) measured in tooth enamel. Although many of the individuals had values consistent with being from Hallstatt culture sites within southwest Germany, some individuals likely originated from further afield. Possible areas include the Alps of Switzerland and Austria or even locations in Italy. Our study strongly supports the assumption of far reaching social and economic networks in the western Hallstatt culture.     

青海喇家遗址人类遗骸的锶同位素比值分析

喇家遗址位于青海省民和县,主体为齐家文化的遗存,距今大约3900~4300年。喇家遗址古代人类迁移活动的研究,对于探索黄河上游古代文明,推动该地区齐家文化研究,有着积极的学术意义。本文首先采用电感耦合等离子体质谱技术对喇家遗址出土的22个个体的44份牙釉质和骨骼样品进行了元素分析,根据检测结果排除了受到污染的2份骨骼样品。其次,通过热电离质谱分析方法对喇家遗址出土22个个体的42份牙釉质和骨骼样品及8个猪牙釉质样品的锶同位素比值进行了测定。检测数据表明,猪牙釉质的锶同位素比值的标准偏差远小于其他动物,可以推断猪由当地饲养的可能性是最大的,因此其牙釉质锶同位素比值可以代表当地的锶同位素水平。经过计算得到8个猪牙釉质的锶同位素比值的平均值为0.710930,将该平均值加或减2倍标准偏差可以得到喇家遗址当地的锶同位素比值范围在0.711058~0.710802之间。以此为前提可以推测,本次检测的22个个体中,有17个个体牙釉质的锶同位素比值在遗址当地的锶同位素比值范围内,可能为本地出生。还有2个个体牙釉质在当地的锶同位素比值范围上下限附近,姑且存疑;仅有3个个体牙釉质的锶同位素比值在遗址当地的锶同位素比值范围以外,表明这些个体可能在其他地区出生,去世后埋葬在遗址中。此外,数据分析还显示,本地出生人群牙釉质的锶同位素比值平均值与骨骼的锶同位素比值平均值比较接近,暗示喇家遗址当地古人类生活方式可能比较统一。     

Migration to the Medieval Middle East with the Crusades

During the 12th and 13th centuries thousands of people moved from Europe to the Middle East to fight, undertake pilgrimage, or settle and make a new life. The aim of this research is to investigate two populations from the Crusader kingdom of Jerusalem, by determining who was born in Europe and who came from the Middle East. Oxygen and strontium stable isotope analyses were conducted on the enamel of teeth from skeletal remains excavated from Crusader contexts. Twenty individuals from the coastal city of Caesarea (10 high status and 10 low status), and two local Near Eastern Christian farmers from the village of Parvum Gerinum (Tel Jezreel) were analyzed as a control sample. Results were compared with known geographic values for oxygen and strontium isotopes. The population of the city of Caesarea appears to have been dominated by European‐born individuals (probably 19/20, but at least 13/20), with few locals. This was surprising as a much higher proportion of locals were expected. Both controls from the farming village of Parvum Gerinum had spent their childhood in the area of the village, which matches our understanding of limited mobility among poor Medieval farmers. This is the first time that stable isotope analysis has been applied to the study of the migration of peoples between Medieval Europe and the Middle East at the time of the crusades. In view of these findings, we must now rethink past estimations of population social structure in Levantine coastal Medieval cities during the Crusader period. Am J Phys Anthropol 2009. © 2009 Wiley‐Liss, Inc.     

Isotopic tracing of the impact of mobility on infectious disease: The origin of people with treponematosis buried in hull,England, in the late medieval period

Treponematosis has been one of the most studied and debated infectious diseases in paleopathology, particularly from the standpoint of its origin, evolution, and transmission. This study links evidence for treponematosis in skeletons from the 14th–16th century AD cemetery of the Augustinian friary of Hull Magistrates Court, England, with data from stable isotope analysis to test the hypothesis that the people with treponemal disease buried at this site were not locally born and raised. The objective is to explore the potential of using stable isotope data to track the place of origin and extent of mobility of individuals with an infectious disease. Dental enamel samples of 12 skeletons were selected for strontium (⁸⁷Sr/⁸⁶Sr ratio) and oxygen (δ¹⁸O) stable isotope analysis based on the presence (six ‐ diseased) or absence (six ‐ controls) of bone changes associated with treponemal disease. The oxygen isotope ratios of all but three individuals (1047, 1121, 823) overlapped at two standard deviations with the inferred local precipitation range, and only one individual (1216) had a strontium isotope ratio outside the regional range. Two of the four had probable/possible treponemal bone changes. Those with treponemal bone changes were not demonstrably more likely to be migrants than those without such lesions. However, because of extensive documentary evidence for trade with the Baltic Sea area, and for merchants from towns such as Stralsund, Danzig and Elbing being in Hull, it is very plausible that the four migrants came from the Baltic area or even southern Sweden. Am J Phys Anthropol, 2013. © 2012 Wiley Periodicals, Inc.     

Changing patterns of mobility as a response to climatic deterioration and aridification in the middle Holocene southern Sahara

The African Humid Period witnessed a rapid human re‐occupation of the Sahara as numerous lakes formed during the Holocene climatic optimum circa 10‐5 kya. Permanent waters attracted a variety of aquatic and terrestrial fauna allowing for long‐term occupation of specific paleolake basins. The Gobero paleolake in central Niger was one such location that preserves a unique mortuary record from the southern Sahara. Here, we use radiogenic strontium isotope analysis to investigate how human communities adapted to aridification throughout the Holocene. In particular, we examine the effects of increasing climate instability on patterns of human mobility. Results of radiogenic strontium isotope analysis of enamel and bone samples from Middle Holocene burials (～7.2–4.9 kya) indicate predominantly local values with no evidence for sex‐based variation. Comparisons of radiogenic strontium isotope data with previously published (Stojanowski and Knudson: Am J Phys Anthropol 146 (2011) 49–61) Early Holocene burials (～9.7–8.3 kya) indicate significant differences in both enamel and bone values. Middle Holocene individuals demonstrate a predominantly non‐local signature for enamel values and a predominantly local signature for bone values. Those individuals with non‐local bone values always demonstrated non‐local enamel values; however, the opposite was not the case. This suggests a divergence of mobility strategies during the Middle Holocene with a minority of individuals maintaining a more mobile existence throughout their life and others maintaining a similar strategy as Early Holocene hunter–gatherers that was tied to the paleolake basin. The more mobile individuals likely lived during the terminal phase of the lake's occupation. One response to aridification by Saharan peoples, then, was increasing mobility. Am J Phys Anthropol 154:79–93, 2014. © 2014 Wiley Periodicals, Inc.     

Utility of multiple chemical techniques in archaeological residential mobility studies: case studies from Tiwanaku- and Chiribaya-affiliated sites in the Andes

In the south central Andes, archaeologists have long debated the extent of Tiwanaku colonization during the Middle Horizon (AD 500-1000). We tested the hypotheses regarding the nature of Tiwanaku influence using strontium isotope, trace element concentration, and oxygen isotope data from archaeological human tooth enamel and bone from Tiwanaku- and Chiribaya-affiliated sites in the south central Andes. Strontium isotope analysis of 25 individuals buried at the Tiwanaku-affiliated Moquegua Valley site of Chen Chen demonstrates that it was likely a Tiwanaku colony. In contrast, no immigrants from the Lake Titicaca Basin were present in 27 individuals analyzed from the San Pedro de Atacama cemeteries of Coyo Oriental, Coyo-3, and Solcor-3; it is likely that these sites represent economic and religious alliances, but not colonies. However, strontium isotope analysis alone cannot distinguish movement between the Tiwanaku- and Chiribaya-affiliated sites in the Moquegua and Ilo Valleys of southern Peru. Analyzing oxygen isotope and trace element concentration data and comparing it with strontium isotope data from the same individuals provides a more detailed picture of residential mobility in the Tiwanaku and Chiribaya polities. In addition to monitoring diagenetic contamination, trace element concentration data identified movement during adulthood for certain individuals. However, these data could not distinguish movement between the Moquegua and Ilo Valleys. While oxygen isotope data could clearly distinguish the high-altitude sites from others, more data is needed to characterize the local oxygen isotope ratios of these regions. These data demonstrate the potential for archaeological reconstruction of residential mobility through multiple lines of evidence.     

Mobility,mortality, and the middle ages: Identification of migrant individuals in a 14th century black death cemetery population

Mobility and migration patterns of groups and individuals have long been a topic of interest to archaeologists, used for broad explanatory models of cultural change as well as illustrations of historical particularism. The 14th century AD was a tumultuous period of history in Britain, with severely erratic weather patterns, the Great Famine of 1315–1322, the Scottish Wars of Independence, and the Hundred Years' War providing additional migration pressures to the ordinary economic issues drawing individuals to their capital under more stable conditions. East Smithfield Black Death Cemetery (Royal Mint) had a documented use period of only 2 years (AD 1348–1350), providing a precise historical context (～50 years) for data. Adults (n = 30) from the East Smithfield site were sampled for strontium and oxygen stable isotope analyses of tooth enamel. Five individuals were demonstrated to be statistical outliers through the combined strontium and oxygen isotope data. Potential origins for migrants ranged from London's surrounding hinterlands to distant portions of northern and western Britain. Historic food sourcing practices for London were found to be an important factor for consideration in a broader than expected ⁸⁷Sr/⁸⁶Sr range reflected in a comparison of enamel samples from three London datasets. The pooled dataset demonstrated a high level of consistency between site data, divergent from the geologically predicted range. We argue that this supports the premise that isotope data in human populations must be approached as a complex interaction between behavior and environment and thus should be interpreted cautiously with the aid of alternate lines of evidence. Am J Phys Anthropol, 2013. © 2012 Wiley Periodicals, Inc.     

Diet, residential origin, and pathology at Machu Picchu, Peru

Pathological conditions in human skeletal remains provide a wealth of information about archaeological populations, but many are limited in their interpretive significance by their nonspecific etiologies. This study analyzes three common pathological conditions known to manifest in infancy and childhood in the skeletal population from Machu Picchu, Peru (N = 74) with published carbon, nitrogen, oxygen, strontium, and lead isotopic data (Turner et al.: J Archaeol Sci 36 (2009) 317–332; Turner et al.: Chungara: Revista de Antropología Chilena 42 (2010) 515–524) to distinguish early‐life diet from residential origins as significantly associated with pathologies among the site's inhabitants. Analyses of variance indicate highly significant variation between enamel δ¹⁸O values, which serve as a rough proxy of local environment, and both cribra orbitalia (CO) and porotic hyperostosis (PH), generally understood to be markers of anemia. Results tentatively suggest that individuals manifesting these lesions may have lived closer to the arid coasts; however, no significant variation was found in parameters of diet (enamel δ¹³C_carbonate, dentin δ¹³C_collagen, dentin δ¹⁵N) by either CO or PH, suggesting that the primary factors causing anemia may have been more significantly related to residential origin rather than diet. Linear enamel hypoplasia (LEH) frequency significantly varied by both dietary and residential parameters, supporting models of LEH formation from a synergy of dietary and environmental factors. These results support previous research on the etiology of PH in the Andes; they also represent a useful approach to refining site‐specific interpretations of pathological conditions in archaeological populations, and exploring etiological variation between populations. Am J Phys Anthropol, 2012. © 2012 Wiley Periodicals, Inc.     

不同产地三疣梭子蟹的稳定同位素比值特征及原产地溯源

探索稳定同位素对三疣梭子蟹产地溯源的可行性,可为保护地理标志产品、追溯原产地来源提供理论依据。本研究以黄海、渤海和东海3个主产区海域的三疣梭子蟹为对象,分析了其碳、氮稳定同位素值差异性和不同组织中稳定同位素比值的相关性。结果表明: 不同产地的碳、氮稳定同位素比值有显著性差异,不同组织之间有明显的同位素分馏效应。利用建立的判别分析模型,通过三疣梭子蟹不同组织稳定同位素比值进行产地判别分析,采用肌肉和腮中碳、氮稳定同位素的产地判别正确率(>95%)明显高于肝胰性腺,说明肌肉和腮稳定同位素比值可以对不同海域的三疣梭子蟹进行有效区分。本研究填补了目前三疣梭子蟹稳定同位素溯源技术的研究空白。     

Isotopic evidence of dietary variations and weaning practices in the Gaya cemetery at Yeanri,Gimhae, South Korea

Stable carbon and nitrogen isotope analyses were conducted to investigate dietary variation in human skeletons (n = 109) from the Gaya cemetery at Yeanri located near Gimhae City, South Korea. The cemetery contained three distinct grave types dating to 4th–7th century AD. The main purposes of this research were to reconstruct palaeodiet in the Gaya population and to explore correlations between stable isotope compositions and burial types, inferred age, and sex of these individuals. The isotopic data indicate that the people at Yeanri consumed a predominantly C₃‐based terrestrial diet supplemented with freshwater and/or marine resources. The comparison of isotopic results reveals significant differences in δ¹³C values among three adult burial types (wood‐cist coffin: ?18.5 ± 0.5‰, stone‐cist coffin: ?18.1 ± 0.6‰, mausoleum: ?17.8 ± 0.9‰). Males in wood‐cist and stone‐cist coffins have relatively more elevated mean δ¹³C and δ¹⁵N values than females. The isotopic ratios from the two adult age groups (21–40 years and 40–60 years) indicate that there was no significant dietary change in individuals with age. The isotope data from the infants and children suggest the weaning was a gradual process that was completed between 3 and 4 years of age in the Gaya population. This evidence indicates that the dietary variations within the cemetery reflect social status, sex, and childhood consumption patterns. Am J Phys Anthropol, 2010. © 2009 Wiley‐Liss, Inc.     

Infant feeding practice in medieval Japan: Stable carbon and nitrogen isotope analysis of human skeletons from Yuigahama‐minami

A longer breastfeeding duration provides various positive effects in subadult health because of abundant immunological factors and nutrients in human breast milk, and decreases the natural fertility of a population through lactational amenorrhea. In this study, we measured stable carbon and nitrogen isotope ratios in the bone collagen of three adults and 45 subadults from the Yuigahama‐minami site (from 12th to 14th century) in Kamakura, the early medieval capital of Japan. Marine foods, C₃‐based terrestrial foods, and freshwater fish are the primarily protein sources for adults. The changes in the nitrogen isotope ratios of subadults suggest that the relative dietary protein contribution from breast milk started to decrease from 1.1 years of age and ended at 3.8 years. The age at the end of weaning in the Yuigahama‐minami population was greater than that in the typical non‐industrial populations, a premodern population in the Edo period Japan, and medieval populations in the UK. Skeletons of townspeople from medieval Kamakura indicate severe nutritional stress (e.g., enamel hypoplasia and cribra orbitalia), yet this longer duration of breastfeeding did not compensate adverse effects for nutritional deficiency. The longer breastfeeding period may have been a consequence of complementary food shortage and bad health of subadults. Kamakura experienced urbanization and population increase in the early medieval period. The younger age‐at‐death distribution and high nutritional stresses in the Yuigahama‐minami population and later weaning, which is closely associated with longer inter‐birth interval for mothers, suggests that Kamakura developed and increased its population by immigration during urbanization. Am J Phys Anthropol 156:241–251, 2015. © 2014 Wiley Periodicals, Inc.     

High individual repeatability and population differentiation in stable isotope ratios in winter-grown collared flycatcher Ficedula albicollis feathers

For migrants, we often lack complete information of their spatial distribution year round. Here, we used stable carbon, nitrogen and hydrogen isotope ratios extracted from feathers grown at the wintering sites of the long-distance migratory collared flycatcher Ficedula albicollis , to study how individuals from different breeding populations are distributed at the wintering sites. A sub-sample of birds was also sampled in two consecutive years to test for the repeatability of isotope ratios. Birds from the same breeding populations had more similar isotope ratios compared to birds from other nearby populations (10–100 km apart). Furthermore, isotope repeatability within individuals was high, implying that the observed pattern of isotope variation is consistent between years. We put forward two hypotheses for these patterns; 1) strong wintering site philopatry and migratory connectivity, suggesting that migratory connectivity may potentially be found on a much smaller spatial scale than previously considered, and 2) consistent interpopulation differentiation of feeding ecology at their wintering site.     

Geographical patterns of human diet derived from stable-isotope analysis of fingernails

Carbon and nitrogen isotope ratios of human fingernails were measured in 490 individuals in the western US and 273 individuals in southeastern Brazil living in urban areas, and 53 individuals living in a moderately isolated area in the central Amazon region of Brazil and consuming mostly locally grown foods. In addition, we measured the carbon and nitrogen isotope ratios of common food items to assess the extent to which these isotopic signatures remain distinct for people eating both omnivorous and vegetarian diets and living in different parts of the world, and the extent to which dietary information can be interpreted from these analyses. Fingernail delta13C values (mean +/- standard deviation) were -15.4 +/- 1.0 and -18.8 +/- 0.8 per thousand and delta15N values were 10.4 +/- 0.7 and 9.4 +/- 0.6 per thousand for southeastern Brazil and western US populations, respectively. Despite opportunities for a "global supermarket" effect to swamp out carbon and nitrogen isotope ratios in these two urbanized regions of the world, differences in the fingernail isotope ratios between southeastern Brazil and western US populations persisted, and appeared to be more associated with regional agricultural and animal production practices. Omnivores and vegetarians from Brazil and the US were isotopically distinct, both within and between regions. In a comparison of fingernails of individuals from an urban city and isolated communities in the Amazonian region, the urban region was similar to southeastern Brazil, whereas individuals from isolated nonurban communities showed distinctive isotopic values consistent with their diets and with the isotopic values of local foods. Although there is a tendency for a "global supermarket" diet, carbon and nitrogen isotopes of human fingernails hold dietary information directly related to both food sources and dietary practices in a region.     

Stable isotope ratios and forensic analysis of microorganisms

In the aftermath of the anthrax letters of 2001, researchers have been exploring various analytical signatures for the purpose of characterizing the production environment of microorganisms. One such signature is stable isotope ratios, which in heterotrophs, are a function of nutrient and water sources. Here we discuss the use of stable isotope ratios in microbial forensics, using as a database the carbon, nitrogen, oxygen, and hydrogen stable isotope ratios of 247 separate cultures of Bacillus subtilis 6051 spores produced on a total of 32 different culture media. In the context of using stable isotope ratios as a signature for sample matching, we present an analysis of variations between individual samples, between cultures produced in tandem, and between cultures produced in the same medium but at different times. Additionally, we correlate the stable isotope ratios of carbon, nitrogen, oxygen, and hydrogen for growth medium nutrients or water with those of spores and show examples of how these relationships can be used to exclude nutrient or water samples as possible growth substrates for specific cultures.     

Isotopic evidence for divergent diets and mobility patterns in the Atacama Desert,northern Chile,during the Late Intermediate Period (AD 900–1450)

The decline of the Tiwanaku state saw the emergence of two new cultures—Pica‐Tarapacá and Atacama—during the Late Intermediate Period in northern Chile. Archeological evidence suggests that both groups practised maize agriculture and pastoralism, but that their interaction zones differed significantly. Marine resources are common at Pica‐Tarapacá sites, even those far from coast, while Atacama sites in the desert oases and precordilleran area seem to have directed their networks towards the highlands. Here we apply stable carbon, nitrogen, and oxygen isotope analysis on human bone and enamel to test dietary patterns and residential mobility at two sites, Pica 8 and Quitor 6, representing the Pica‐Tarapacá and Atacama cultures, respectively. Our results show that diet at the two sites indeed differed: significant but variable consumption of marine resources and maize is indicated at Pica 8, despite being an inland site, while diet at Quitor 6 was based mainly on terrestrial resources. The use of seabird guano and llama dung as fertilizers and extreme aridity may have contributed to the high nitrogen isotope values observed in Pica 8 humans. The δ¹⁸O values in Pica 8 individuals are generally lower than for Quitor in spite of its greater distance from the Andes. All three isotopes suggest the presence of at least five nonlocals in the 30 measured at Pica 8. This evidence for human mobility is consistent with the high levels of trade and interaction observed in the archeological record, and begins to quantify the degree of movement of specific individuals. Am J Phys Anthropol 156:374–387, 2015. © 2014 Wiley Periodicals, Inc.     

Implications of tooth root hypercementosis in a Barbados slave skeletal collection

A 17th- to 19th-century cemetery sample of 104 slaves from Newton Plantation (Barbados) shows uniquely high hypercementosis prevalence, as well as unexpectedly high and variable skeletal lead content. A variety of biological and archeological factors indicates that individuals with lower amounts of these anomalies (relative to age at death) were probably African-born, first-generation slaves. The hypercementosis is related to the progression of periodontal disease as assessed from alveolar bone. Although the hypercementosis is endemic in the Caribbean black population, it does not as yet have a clear explanation. We suggest the etiology might relate to chronic malnutrition involving periodic, seasonal rehabilitation.     

Childhood lead exposure in an enslaved African community in Barbados: Implications for birthplace and health status

Lead was ubiquitous on Caribbean sugar plantations, where it was used extensively in the production of sugar and rum. Previous studies suggest that skeletal lead contents can be used to identify African‐born individuals (as opposed to Creoles) among slave burials found in the New World. To test this hypothesis, we measured lead concentrations in enamel samples from 26 individuals from the Newton Plantation cemetery in Barbados, which was in use from around 1660 to 1820, and compared the results with enamel ⁸⁷Sr/⁸⁶Sr measurements that had been previously obtained for the same population. Results show a clear association between low (i.e., below 1 ppm) enamel lead concentrations and higher enamel ⁸⁷Sr/⁸⁶Sr ratios which have previously been interpreted as being indicative of African birth, suggesting that individuals with low enamel lead levels were indeed born in Africa as opposed to the New World. Based on these results, we propose that enamel lead measurements provide an effective and inexpensive way to determine African birth from skeletal remains. Furthermore, the lead measurements can provide useful insights into the health status and childhood environment of enslaved Africans during the colonial period. Am J Phys Anthropol, 2013. © 2012 Wiley Periodicals, Inc.     

Stable Isotope Ratios and Forensic Analysis of Microorganisms

In the aftermath of the anthrax letters of 2001, researchers have been exploring various analytical signatures for the purpose of characterizing the production environment of microorganisms. One such signature is stable isotope ratios, which in heterotrophs, are a function of nutrient and water sources. Here we discuss the use of stable isotope ratios in microbial forensics, using as a database the carbon, nitrogen, oxygen, and hydrogen stable isotope ratios of 247 separate cultures of Bacillus subtilis 6051 spores produced on a total of 32 different culture media. In the context of using stable isotope ratios as a signature for sample matching, we present an analysis of variations between individual samples, between cultures produced in tandem, and between cultures produced in the same medium but at different times. Additionally, we correlate the stable isotope ratios of carbon, nitrogen, oxygen, and hydrogen for growth medium nutrients or water with those of spores and show examples of how these relationships can be used to exclude nutrient or water samples as possible growth substrates for specific cultures.