Isotopic evidence for age-related immigration to imperial Rome

Oxygen stable isotope ratios (delta(18)O) have been determined in carbonate in paired first and third molar teeth from individuals (N = 61) who lived in the town of Portus Romae ("Portus") and who were buried in the necropolis of Isola Sacra (First to Third centuries AD) near Rome, Italy. We compare these analyses with data for deciduous teeth of modern Roman children. Approximately one-third of the archaeological sample has first molar (M1) values outside the modern range, implying a large rate of population turnover at that time, consistent with historical data. Delta (18)O(ap) values suggest that a group within the sample migrated to the area before the third molar (M3) crown had completely formed (i.e., between 10 and 17.5 years of age). This is the first quantitative assessment of population mobility in Classical antiquity. This study demonstrates that migration was not limited to predominantly single adult males, as suggested by historical sources, but rather a complex phenomenon involving families. We hypothesize that migrants most likely came from higher elevations to the East and North of Rome. One individual with a higher delta(18)O value may have come (as a child) from an area isotopically similar to North Africa.     

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.     

Isotopic evidence for breastfeeding and possible adult dietary differences from Late/Sub-Roman Britain

Historical documents indicate that breastfeeding and weaning practices have fluctuated in England through history. In order to obtain evidence for general breastfeeding patterns in Late/Sub-Roman Britain, stable carbon and nitrogen isotope values were measured in juvenile and adult skeletons (n = 87) from the cemetery of Queenford Farm, Dorchester-on-Thames, Oxfordshire. As the site contained few individuals between 0-1.5 years of age, it was not possible to determine the initial timing for the introduction of weaning foods. Between ages 2-4 years, the mean +/- SD delta(13)C results (-20.2 +/- 0.3 per thousand) are significantly more negative (t = -4.03, P < 0.001) compared to adult females (-19.7 +/- 0.3 per thousand). This is interpreted as evidence of a different diet being fed to children during weaning. After age 2, the delta(15)N values gradually decline, indicating complete cessation of breastfeeding by 3-4 years. Among adults, stature (males = 1.68 +/- 0.06 m; females = 1.58 +/- 0.07 m) and sexual dimorphism (106) were low, suggesting that the population was possibly under environmental stress. The delta(13)C results for adults are similar, but females show a small but statistically significantly (t = -2.86, P < 0.01) lower mean delta(15)N value (9.9 +/- 0.9 per thousand) compared to males (10.6 +/- 0.5 per thousand). These lower female delta(15)N values possibly reflect the different physiology of the sexes (pregnancy and/or lactation) or the reduced consumption of animal/fish protein by women, and this may have been influenced by individual preference, family needs, or societal values of the era.     

Isotopic and genetic evidence for culturally inherited site fidelity to feeding grounds in southern right whales (Eubalaena australis)

Ocean warming will undoubtedly affect the migratory patterns of many marine species, but specific changes can be predicted only where behavioural mechanisms guiding migration are understood. Southern right whales show maternally inherited site fidelity to near-shore winter nursery grounds, but exactly where they feed in summer (collectively and individually) remains mysterious. They consume huge quantities of copepods and krill, and their reproductive rates respond to fluctuations in krill abundance linked to El Niño Southern Oscillation (ENSO). Here we show that genetic and isotopic signatures, analysed together, indicate maternally directed site fidelity to diverse summer feeding grounds for female right whales calving at Península Valdés, Argentina. Isotopic values from 131 skin samples span a broad range (–23.1 to –17.2‰δ¹³C, 6.0 to 13.8‰δ¹⁵N) and are more similar than expected among individuals sharing the same mitochondrial haplotype. This pattern indicates that calves learn summer feeding locations from their mothers, and that the timescale of culturally inherited site fidelity to feeding grounds is at least several generations. Such conservatism would be expected to limit the exploration of new feeding opportunities, and may explain why this population shows increased rates of reproductive failure in years following elevated sea-surface temperature anomalies off South Georgia, the richest known feeding ground for baleen whales in the South Atlantic.     

The variable roads to sacrifice: Isotopic investigations of human remains from Chotuna‐Huaca de los Sacrificios,Lambayeque, Peru

This study investigates two key variables—residential context and subsistence—among sacrificial victims dating to the Late Horizon (A.D. 1450–1532) in the Huaca de los Sacrificios at the Chotuna‐Chornancap Archaeological Complex in north coastal Peru. We investigate whether aspects of sacrifice in this distant coastal province mirrored that found in Inca heartland contexts such as the capacocha, or remained more typical of coastal sacrificial traditions. Stable carbon, nitrogen, and oxygen isotope values were characterized in bone carbonate, bone collagen, and hair keratin to estimate geographic residence during the decade before death and diet in the decade, versus months, before death. Bone δ¹⁸O_carbonate values have a mean (±SD) of 26.8 ± 1.1%, bone δ¹³C_carbonate values ?6.7 ± 1.7%, and bone δ¹³C_collagen values 11.8 ± 1.3%; bone δ¹⁵N_collagen values have a mean of 11.5 ± 1.3%. Combined hair δ¹³C_keratin values have a mean of ?12.8 ± 1.6%, and hair δ¹⁵N_keratin values 10.8 ± 1.3%. In contrast to contemporaneous coastal and highland contexts, we are unable to identify immigrants among the sacrificed individuals or changes in diet that indicate provisioning with a standardized diet leading up to death. Instead, results suggest that victims were local to the area, but consumed moderately variable diets consistent with local subsistence patterns. These findings suggest a distinct pattern of human sacrifice in the Late Horizon and underscore the regional and temporal variation in sacrificial practices in the central Andes. Am J Phys Anthropol 151:22–37, 2013. © 2013 Wiley Periodicals, Inc.     

Consistency of elemental and isotope‐ratio patterns across multiple scales from individual fish

Assays of elemental and stable‐isotope ratios across growth increments of scales have the potential to provide a non‐lethal alternative to otolith chemistry for identifying migration and ontogenetic trophic shifts. A central assumption when employing scales as otolith analogues is that any scale from an individual will provide equivalent information about the chemical history of that fish. This assumption was investigated with multiple scales from wild and captive euryhaline Atlantic tarpon Megalops atlanticus from the north‐west Gulf of Mexico. Elemental (Sr:Ca) and isotope‐ratio (δ¹³C and δ¹⁵N) life‐history profiles were compared among multiple scales from each fish. All three chemical proxies showed highly consistent patterns among non‐regenerated scales, while patterns in regenerated scales diverged, indicating rapid regrowth of interior scale material at the onset of regeneration. Patterns of Sr:Ca and δ¹³C covaried, supporting their use as salinity proxies, while δ¹⁵N patterns were consistent with ontogenetic diet shifts. Water samples taken from aquaria holding captive fish were used to calculate partition coefficients for a suite of elements in M. atlanticus scales for future quantification of migratory movements in the region. Together, these results support the assumption that non‐regenerated scales from individual M. atlanticus provide equivalent chemical histories, further validating their use as a viable non‐lethal alternative to otoliths.     

Aquatic food‐web dynamics following incorporation of nutrients derived from Atlantic anadromous fishes

Changes in the isotopic composition (δ¹³C and δ¹⁵N) in biofilm, macro‐invertebrates and resident salmonids were used to characterize temporal dynamics of marine derived nutrients (MDNs) incorporation between stream reaches with and without MDN inputs. Five Atlantic rivers were chosen to represent contrasting MDN subsidies: four rivers with considerable numbers of anadromous fishes; one river with little MDN input. Rainbow smelt Osmerus mordax, alewife Alosa pseudoharengus, sea lamprey Petromyzon marinus and Atlantic salmon Salmo salar, were the primary anadromous species for the sampled rivers. Regardless of the spatial resolution or the pathway of incorporation, annual nutrient pulses from spawning anadromous fishes had a positive effect on isotopic enrichment at all trophic levels (biofilm, 1·2–5·4‰; macro‐invertebrates, 0·0–6·8‰; fish, 1·2–2·6‰). Community‐wide niche space shifted toward the marine‐nutrient source, but the total ecological niche space did not always increase with MDN inputs. The time‐integrated marine‐nutrient resource contribution to the diet of S. salar parr and brook trout Salvelinus fontinalis ranged between 16·3 and 36·0% during anadromous fish‐spawning periods. The high degree of spatio‐temporal heterogeneity in marine‐nutrient subsidies from anadromous fishes lead to both direct and indirect pathways of MDN incorporation into stream food webs. This suggests that organisms at many trophic levels derive a substantial proportion of their energy from marine resources when present. The current trend of declining anadromous fish populations means fewer nutrient‐rich marine subsidies being delivered to rivers, diminishing the ability to sustain elevated riverine productivity.     

A Liquid Chromatography–Mass Spectrometry Method to Measure Stable Isotopic Tracer Enrichments of Glycerol and Glucose in Human Serum

Stable isotopes are commonly used as tracers for the measurement of glycerol and glucose kinetics in metabolic studies. Traditionally, the analysis of these isotopes has been performed using gas chromatography-mass spectrometry, which requires that the analytes first be derivatized. The derivatization process adds considerable complexity to the method. Liquid chromatography-mass spectrometry (LCMS) can measure many metabolites directly with limited sample preparation. We present a novel analytical method for the measurement of [1,1,2,3,3-(2)H(5)]glycerol (d(5)-glycerol) and [6,6-(2)H(2)]glucose (d(2)-glucose) isotopic tracer enrichments in human serum in a single run by LCMS. After a simple extraction step, the sample is separated isocratically by HPLC, and the isotopes are measured using positive electrospray ionization with selected ion monitoring of the sodium-adduct ions. The method is linear over a wide range of d(2)-glucose and d(5)-glycerol enrichments. The within-day standard deviation of measurement of serum samples was 0.05 mole% excess (MPE) for d(2)-glucose and 0.25 MPE for d(5)-glycerol. The variation of tracer enrichment among days was about double that measured within 1 day.     

Pronghorn (Antilocapra americana) enamel phosphate δ18O values reflect climate seasonality: Implications for paleoclimate reconstruction

Stable oxygen isotope (δ¹⁸O) compositions from vertebrate tooth enamel are widely used as biogeochemical proxies for paleoclimate. However, the utility of enamel oxygen isotope values for environmental reconstruction varies among species. Herein, we evaluate the use of stable oxygen isotope compositions from pronghorn (Antilocapra americana Gray, 1866) enamel for reconstructing paleoclimate seasonality, an elusive but important parameter for understanding past ecosystems. We serially sampled the lower third molars of recent adult pronghorn from Wyoming for δ¹⁸O in phosphate (δ¹⁸O_PO4) and compared patterns to interpolated and measured yearly variation in environmental waters as well as from sagebrush leaves, lakes, and rivers (δ¹⁸O_w). As expected, the oxygen isotope compositions of phosphate from pronghorn enamel are enriched in ¹⁸O relative to environmental waters. For a more direct comparison, we converted δ¹⁸O_w values into expected δ¹⁸O_PO4* values (δ¹⁸O_W‐_PO4*). Pronghorn δ¹⁸O_PO4 values from tooth enamel record nearly the full amplitude of seasonal variation from Wyoming δ¹⁸O_W‐PO4* values. Furthermore, pronghorn enamel δ¹⁸O_PO4 values are more similar to modeled δ¹⁸O_W‐PO4* values from plant leaf waters than meteoric waters, suggesting that they obtain much of their water from evaporated plant waters. Collectively, our findings establish that seasonality in source water is reliably reflected in pronghorn enamel, providing the basis for exploring changes in the amplitude of seasonality of ancient climates. As a preliminary test, we sampled historical pronghorn specimens (1720 ± 100 AD), which show a mean decrease (a shift to lower values) of 1–2‰ in δ¹⁸O_PO4 compared to the modern specimens. They also exhibit an increase in the δ¹⁸O amplitude, representing an increase in seasonality. We suggest that the cooler mean annual and summer temperatures typical of the 18th century, as well as enhanced periods of drought, drove differences among the modern and historical pronghorn, further establishing pronghorn enamel as excellent sources of paleoclimate proxy data.     

Sensitivity of south‐east Atlantic planktonic foraminifera to mid‐Pleistocene climate change

Abstract: The last one million years are important in terms of climate development during the so‐called Mid‐Pleistocene Transition when amplification of the glacial–interglacial cycles occurred. This study describes abundance changes in fossil planktonic foraminifera in sediments from Core T89‐40, retrieved from the Walvis Ridge in the south‐east Atlantic, across this time period. Cycles between upwelling and subtropical planktonic foraminiferal assemblages are shown to mirror changes between glacial and interglacial periods, respectively. During interglacial marine isotopic stages (MIS) 9, 11 and 31, however, anomalously high abundances of the polar left‐coiled Neogloboquadrina pachyderma occur, presumably linked to unusual seasonal upwelling waters. The planktonic foraminiferal abundance record shows 41‐ky cyclic variations in the regional oceanography linked to cycles in insolation influenced by changes in the Earth’s axial tilt (obliquity). These orbitally induced oscillations in oceanographic change occurred throughout the entire record. The most conspicuous feature of the planktonic foraminiferal record is the near absence of left‐coiled Globorotalia truncatulinoides between 960 and 610 ka (MIS 26‐15). The abrupt disappearance of this species is synchronous with the onset of the Mid‐Pleistocene Transition in MIS 26.     

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.     

Isotopic niche differs between seal and fish‐eating killer whales (Orcinus orca) in northern Norway

Ecological diversity has been reported for killer whales (Orcinus orca) throughout the North Atlantic but patterns of prey specialization have remained poorly understood. We quantify interindividual dietary variations in killer whales (n = 38) sampled throughout the year in 2017–2018 in northern Norway using stable isotopic nitrogen (δ¹⁵N: ¹⁵N/¹⁴N) and carbon (δ¹³C: ¹³C/¹²C) ratios. A Gaussian mixture model assigned sampled individuals to three differentiated clusters, characterized by disparate nonoverlapping isotopic niches, that were consistent with predatory field observations: seal‐eaters, herring‐eaters, and lumpfish‐eaters. Seal‐eaters showed higher δ¹⁵N values (mean ± SD: 12.6 ± 0.3‰, range = 12.3–13.2‰, n = 10) compared to herring‐eaters (mean ± SD: 11.7 ± 0.2‰, range = 11.4–11.9‰, n = 19) and lumpfish‐eaters (mean ± SD: 11.6 ± 0.2‰, range = 11.3–11.9, n = 9). Elevated δ¹⁵N values for seal‐eaters, regardless of sampling season, confirmed feeding at high trophic levels throughout the year. However, a wide isotopic niche and low measured δ¹⁵N values in the seal‐eaters, compared to that of whales that would eat solely seals (δ_N‐measured = 12.6 vs. δ_N‐expected = 15.5), indicated a diverse diet that includes both fish and mammal prey. A narrow niche for killer whales sampled at herring and lumpfish seasonal grounds supported seasonal prey specialization reflective of local peaks in prey abundance for the two fish‐eating groups. Our results, thus, show differences in prey specialization within this killer whale population in Norway and that the episodic observations of killer whales feeding on prey other than fish are a consistent behavior, as reflected in different isotopic niches between seal and fish‐eating individuals.     

Long‐term increases in intrinsic water‐use efficiency do not lead to increased stem growth in a tropical monsoon forest in western Thailand

Rising atmospheric carbon dioxide [CO₂] can accelerate tree growth by stimulating photosynthesis and increasing intrinsic water‐use efficiency (iWUE). Little evidence exists, however, for the long‐term growth and gas‐exchange responses of mature trees in tropical forests to the combined effects of rising [CO₂] and other global changes such as warming. Using tree rings and stable isotopes of carbon and oxygen, we investigated long‐term trends in the iWUE and stem growth (basal area increment, BAI) of three canopy tree species in a tropical monsoon forest in western Thailand (Chukrasia tabularis, Melia azedarach, and Toona ciliata). To do this, we modelled the contribution of ontogenetic effects (tree diameter or age) and calendar year to variation in iWUE, oxygen isotopes, and BAI using mixed‐effects models. Although iWUE increased significantly with both tree diameter and calendar year in all species, BAI at a given tree diameter was lower in more recent years. For one species, C. tabularis, differences in crown dominance significantly influence stable isotopes and growth. Tree ring Δ¹⁸O increased with calendar year in all species, suggesting that increasing iWUE may have been driven by relatively greater reductions in stomatal conductance – leading to enrichment in Δ¹⁸O – than increases in photosynthetic capacity. Plausible explanations for the observed declines in growth include water stress resulting from rising temperatures and El Niño events, increased respiration, changes in allocation, or more likely, a combination of these factors.     

Beyond carbon and nitrogen: guidelines for estimating three‐dimensional isotopic niche space

Isotopic niche has typically been characterized through carbon and nitrogen ratios and most modeling approaches are limited to two dimensions. Yet, other stable isotopes can provide additional power to resolve questions associated with foraging, migration, dispersal and variations in resource use. The ellipse niche model was recently generalized to n‐dimensions. We present an analogous methodology which incorporates variation across three stable dimensions to estimate the significant features of a population's isotopic niche space including: 1) niche volume (referred to as standard ellipsoid volume, SEV), 2) relative centroid location (CL), 3) shape and 4) area of overlap between multiple ellipsoids and 5) distance between two CLs. We conducted a simulation study showing the accuracy and precision of three dimensional niche models across a range of values. Importantly, the model correctly identifies differences in SEV and CL among populations, even with small sample sizes and in cases where the absolute values cannot precisely be recovered. We use these results to provide guidelines for sample size in conducting multivariate isotopic niche modeling. We demonstrate the utility of our approach with a case study of three bottlenose dolphin populations which appear to possess largely overlapping niches when analyzed with only carbon and nitrogen isotopes. Upon inclusion of sulfur, we see that the three dolphin ecotypes are in fact segregated on the basis of salinity and find the stable isotope niche of inshore bottlenose dolphins significantly larger than coastal and offshore populations.     

Trans‐equatorial migration,staging sites and wintering area of Sabine’s Gulls Larus sabini in the Atlantic Ocean

The migrations and winter distributions of most seabirds, particularly small pelagic species, remain poorly understood despite their potential as indicators of marine ecosystem health. Here we report the use of miniature archival light loggers (geolocators) to track the annual migration of Sabine’s Gull Larus sabini, a small (c. 200 g) Arctic‐breeding larid. We describe their migratory routes and identify previously unknown staging sites in the Atlantic Ocean, as well as their main Atlantic wintering area in the southern hemisphere. Sabine’s Gulls breeding in northeast Greenland displayed an average annual migration of almost 32 000 km (n = 6), with the longest return journey spanning close to 39 000 km (not including local movements at staging sites or within the wintering area). On their southern migration, they spent an average of 45 days in the Bay of Biscay and Iberian Sea, off the coasts of France, Spain and Portugal. They all wintered in close association with the cold waters of the Benguela Upwelling, spending an average of 152 days in that area. On their return north, Sabine’s Gulls staged off the west African coast (Morocco, Mauritania, Senegal), spending on average 19 days at this site. This leg of migration was particularly rapid, birds travelling an average of 813 km/day, assisted by the prevailing winds. Sabine’s Gulls generally followed a similar path on their outbound and return migrations, and did not exhibit the broad figure‐of‐eight pattern (anti clockwise in the southern hemisphere and clockwise in the northern hemisphere) seen in other trans‐equatorial seabirds in the Atlantic and Pacific oceans.     

Southern French Neolithic populations: Isotopic evidence for regional specificities in environment and diet

The Middle Neolithic of the Northwestern Mediterranean area (～4500–3500 BC cal) is characterized by the development of food production techniques as well as by increasing social complexity. These characteristics could have had an impact on human dietary patterns. To evaluate human dietary practices and lifeways of the Middle Neolithic populations from the South of France, stable carbon and nitrogen isotope analysis was carried out on 57 human and 53 faunal bones from seven archaeological sites located in the Languedoc and Garonne regions between 20 and 100 km from the Mediterranean Sea, respectively. Results show regional differences in carbon isotope values. Animal and human bones from the Languedoc region are significantly enriched in ¹³C relative to the Garonne. Conversely, human and dog bones from the Garonne region are significantly enriched in ¹⁵N compared to human and dog bones from the Languedoc region. These results highlight the importance of the local ecosystem in human and animal diet as well as a regional differentiation of palaeodietary behavior, which probably relates to economic and social factors. The comparison of stable isotope data with archaeological and biological evidence does not show any significant intra‐ or interpopulation differences. However, the presence of human outliers suggests that migration probably occurred, perhaps in relation to the trade of animals and/or materials. This study also highlights the importance of investigating local animal stable isotope values for the interpretation of human palaeodiet. Am J Phys Anthropol 2010. © 2009 Wiley‐Liss, Inc.     

Paleoceanography of the Late Cretaceous (Maastrichtian) Western Interior Seaway of North America: evidence from Sr and O isotopes

Well-preserved fossils of the Late Cretaceous Western Interior Seaway (WIS) of North America have been analyzed for Sr concentration and Sr and O isotopes in order to decipher paleosalinities and paleotemperatures. The samples are from four biofacies within the Seaway (late Maastrichtian): offshore Interior (Pierre Shale), nearshore Interior (Fox Hills Formation), brackish (reduced salinity; Fox Hills Formation) and freshwater (Hell Creek Formation). Samples were also obtained from the Severn Formation of Maryland (considered to be representative of the open ocean). All biofacies (except the freshwater) are demonstrably within the Jeletzkytes nebrascensis ammonite zone (<1 Ma duration). The ⁸⁷Sr/⁸⁶Sr ratios show significant and systematic decreases from marine (mean±1 S.D.=0.707839±0.000024) to brackish facies (mean±1 S.D.=0.707677±0.000036), consistent with dilution by freshwater with a lower ⁸⁷Sr/⁸⁶Sr ratio than seawater. Such variation disallows using the ⁸⁷Sr/⁸⁶Sr ratios of fossil shell material to assign ages to fossils from the Late Cretaceous WIS without knowledge of the salinity in which the organism grew. The Sr isotope ratios for scaphitid ammonites within a single biofacies are similar to each other and different from those for scaphites in other biofacies, implying that these organisms are restricted in their distribution during life. The ⁸⁷Sr/⁸⁶Sr values of freshwater unionid mussels range widely and are not compatible with the freshwater endmember ⁸⁷Sr/⁸⁶Sr ratio required by the trend in ⁸⁷Sr/⁸⁶Sr vs. biofacies established from the other samples. Paleosalinities for the biofacies are estimated to range from 35‰ in the open marine to a minimum of 20‰ in the brackish, based on the presence of cephalopods in all four facies and the known salinity tolerance of modern cephalopods. Producing reasonable ⁸⁷Sr/⁸⁶Sr values for the freshwater endmember of a ⁸⁷Sr/⁸⁶Sr vs. 1/[Sr] plot requires a Sr concentration 0.2-0.5 that of seawater for the dominant freshwater input to the WIS. Such high Sr concentrations (relative to seawater) are not observed in modern rivers, and we suggest that the brackish environment in the WIS arose through the mixing of freshwater and seawater in a nearshore aquifer system. Reactions of the solution with aquifer solids in this ‘subterranean estuary’ [Moore, Mar. Chem. 65 (1999) 111-125] produced brackish water with the Sr concentration and isotopic composition recorded in the brackish biofacies. δ¹⁸O values of the fossils show decreases from the marine to brackish biofacies consistent with increasing temperatures (from ∼13 to 23°C) or, if temperatures were relatively constant, to a decrease in the δ¹⁸O of the water in which the shell formed. The latter interpretation is consistent with less-than-fully marine salinities in the nearshore biofacies, but both changes in temperature and the isotopic composition of the water may have occurred in this environment.