Investigating infant feeding development in wild chimpanzees using stable isotopes of naturally shed hair.

Measuring the relative contributions of milk and non-milk foods in the diets of primate infants is difficult from observations. Stable carbon (δ13 C) and nitrogen (δ15 N) isotopes in hair can be used to physiologically track infant feeding through development, but few wild studies have done so, likely due to the difficulty in collecting hair non-invasively. We assessed infant feeding at different ages in wild chimpanzees (Pan troglodytes) at Ngogo, Uganda using δ13 C and δ15 N of keratin in 164 naturally shed hairs from 29 infants (61 hairs), 6 juveniles (7 hairs), 28 mothers (67 hairs) and 14 adult males (29 hairs). Hairs were collected when they stuck to feces during defecation or from the ground after chimpanzees groomed or rested. We could not distinguish between the hairs of infants and mothers using strand length and diameter. Infants 1-2 years old were most enriched in 13 C and 15 N and showed means of 1.1‰ in δ13 C and 2.1‰ in δ15 N above their mothers. Infants at 2 years had hair δ13 C values like those of their mothers, which suggests infants began relying more heavily on plants around this age. While mother-infant δ13 C and δ15 N differences generally decreased with offspring age, as is expected when infants rely increasingly more on independent foraging through development, milk seemed to remain an important dietary component for infants older than 2.5 years, as evidenced by continuing elevated δ15 N. We showed that stable carbon and nitrogen isotopes in naturally shed hairs can feasibly detect trophic level differences between chimpanzee infants and mothers. Since it can mitigate some of the limitations associated with behavioral and fecal stable isotope data, the use of hair stable isotopes is a useful, non-invasive tool for assessing infant feeding development in wild primates.

Successful reconstruction of whole mitochondrial genomes from ancient Central America and Mexico.

The northern and southern peripheries of ancient Mesoamerica are poorly understood. There has been speculation over whether borderland cultures such as Greater Nicoya and Casas Grandes represent Mesoamerican outposts in the Isthmo-Colombian area and the Greater Southwest, respectively. Poor ancient DNA preservation in these regions challenged previous attempts to resolve these questions using conventional genetic techniques. We apply advanced in-solution mitogenome capture and high-throughput sequencing to fourteen dental samples obtained from the Greater Nicoya sites of Jícaro and La Cascabel in northwest Costa Rica (n = 9; A.D. 800-1250) and the Casas Grandes sites of Paquimé and Convento in northwest Mexico (n = 5; A.D. 1200-1450). Full mitogenome reconstruction was successful for three individuals from Jícaro and five individuals from Paquimé and Convento. The three Jícaro individuals belong to haplogroup B2d, a haplogroup found today only among Central American Chibchan-speakers. The five Paquimé and Convento individuals belong to haplogroups C1c1a, C1c5, B2f and B2a which, are found in contemporary populations in North America and Mesoamerica. We report the first successfully reconstructed ancient mitogenomes from Central America, and the first genetic evidence of ancestry affinity of the ancient inhabitants of Greater Nicoya and Casas Grandes with contemporary Isthmo-Columbian and Greater Southwest populations, respectively.

Ancient mitochondrial DNA and ancestry of Paquimé inhabitants, Casas Grandes (A.D. 1200-1450).

OBJECTIVES: The Casas Grandes (Paquimé) culture, located in the Northwest of Chihuahua, Mexico reached its apogee during the Medio Period (A.D. 1200-1450). Paquimé was abandoned by the end of the Medio Period (A.D. 1450), and the ancestry of its inhabitants remains unsolved. Some authors suggest that waves of Mesoamerican immigrants, possibly merchants, stimulated Paquimé's development during the Medio Period. Archaeological evidence suggests possible ties to groups that inhabited the Southwestern US cultures. This study uses ancient DNA analysis from fourteen samples to estimate genetic affinities of ancient Paquimé inhabitants. MATERIALS AND METHODS: DNA was extracted from 14 dental ancient samples from Paquimé. PCR and Sanger sequencing were used to obtain mitochondrial control region sequences. Networks, PCoA, and Nei genetic distances were estimated to compare Paquimé haplotypes against available past haplotypes data from Southwestern and Mesoamerican groups. RESULTS: Haplogroups were characterized for 11 of the samples, and the results revealed the presence of four distinct Amerindian mitochondrial lineages: B (n = 5; 45%), A (n = 3; 27%), C (n = 2; 18%) and D (n = 1; 10%). Statistical analysis of the haplotypes, haplogroup frequencies, and Nei genetic distances showed close affinity of Paquimé with Mimbres. DISCUSSION: Although our results provide strong evidence of genetic affinities between Paquimé and Mimbres, with the majority of haplotypes shared or derived from ancient Southwest populations, the causes of cultural development at Paquimé still remain a question. These preliminary results provide evidence in support of other bioarchaeological studies, which have shown close biological affinities between Paquimé and Mimbres, a Puebloan culture, in the Southwestern US.

A novel fecal stable isotope approach to determine the timing of age-related feeding transitions in wild infant chimpanzees.

OBJECTIVES: Determining nutritional development in wild primates is difficult through observations because confirming dietary intake is challenging. Physiological measures are needed to determine the relative contributions of maternal milk and other foods at different ages, and time of weaning. We used fecal stable carbon and nitrogen isotopes (δ13 C, δ15 N) and fecal nitrogen concentrations (%N) from wild chimpanzees at Ngogo, Uganda, to derive physiological dietary indicators during the transition from total reliance on maternal milk to adult foods after weaning. MATERIALS AND METHODS: We analyzed 560 fecal samples collected non-invasively from 48 infants, their mothers, and 6 juvenile siblings. Most infant and juvenile samples (90%) were matched to samples collected from mothers on the same day. Isotopic assessments were compared with observations of nursing and feeding. RESULTS: Infants ≤1 year old showed average δ15 N, δ13 C and %N ratios that were 2.0‰, 0.8‰ and 1.3% greater than their mothers, respectively, interpreted as trophic level effects. Although data collected on newborns were few, results suggest that solid foods were consumed within 2-5 months after birth. Trophic level differences decreased steadily after 1 year, which indicates a decreasing relative contribution of milk to the diet. Isotopic results indicated infants were weaned by 4.5 years old-more than a year earlier than observations of nipple contacts ceased, which revealed the occurrence of "comfort nursing." Juvenile isotopic signatures indicate no nursing overlap between siblings. DISCUSSION: Our results resemble the stable isotope differences of human babies. This study contributes to a model of chimpanzee nutritional development required to understand early life history patterns in hominins.

Alloparenting is associated with reduced maternal lactation effort and faster weaning in wild chimpanzees.

Alloparenting, when individuals other than the mother assist with infant care, can vary between and within populations and has potential fitness costs and benefits for individuals involved. We investigated the effects of alloparenting on the speed with which infants were weaned, a potential component of maternal fitness because of how it can affect inter-birth intervals, in wild chimpanzees (Pan troglodytes schweinfurthii) at Ngogo, Uganda. We also provide, to our knowledge, the first description of alloparenting in this population and present a novel measure of the contribution of milk to infant diets through faecal stable nitrogen isotopes (δ15N). Using 42 mother-infant pairs, we tested associations of two alloparenting dimensions, natal attraction (interest in infants) and infant handling (holding, carrying), to the proportion of time mothers spent feeding and to maternal lactation effort (mean nursing rates and mother-infant δ15N differences). Neither natal attraction nor infant handling was significantly associated with feeding time. Infant handling was inversely associated with both measures of lactation effort, although natal attraction showed no association. Alloparenting may benefit mothers by enabling females to invest in their next offspring sooner through accelerated weaning. Our findings emphasize the significance of alloparenting as a flexible component of female reproductive strategies in some species.

Bone quality in prehistoric, Cis-Baikal forager femora: a micro-CT analysis of cortical canal microstructure.

Bone quality, a contributor to bone strength, is determined by structural and mechanical properties, which may be analyzed by gross and/or microscopic methods. Variables that contribute to bone quality, such as porosity, can provide insight into the health and lifestyles of people in prehistory. This study tests the ability of microcomputed tomography (µCT) to capture and characterize cortical canal systems in archaeological bone. Seven variables and 71 femora are analyzed to explore bone dynamics in prehistoric foragers from Lake Baikal, Siberia. The results indicate that canal number and canal separation differ significantly (P < 0.05) between age-at-death categories, but only for the pooled and male samples. When merged into a new variable by means of principal components analysis, canal diameter and canal surface to canal volume are also able to discriminate amongst age-at-death categories, as well as between the sexes. However, the overall lack of significant differences between the sexes and amongst age-at-death categories indicates that Baikal forager bone quality (i.e., canal architecture) did not change drastically throughout the lifespan. Interestingly, principal component one identified an untested variable that contributes to canal microstructure variability, and a sexual division of labor may promote divergent trends in canal degree of anisotropy between the sexes. Overall, µCT provides an alternate method for exploring bone quality in archaeological remains, complementing existing methods such as thin-sectioning and gross morphological analyses.

Burying dogs in ancient Cis-Baikal, Siberia: temporal trends and relationships with human diet and subsistence practices.

The first objective of this study is to examine temporal patterns in ancient dog burials in the Lake Baikal region of Eastern Siberia. The second objective is to determine if the practice of dog burial here can be correlated with patterns in human subsistence practices, in particular a reliance on terrestrial mammals. Direct radiocarbon dating of a suite of the region's dog remains indicates that these animals were given burial only during periods in which human burials were common. Dog burials of any kind were most common during the Early Neolithic (∼7-8000 B.P.), and rare during all other time periods. Further, only foraging groups seem to have buried canids in this region, as pastoralist habitation sites and cemeteries generally lack dog interments, with the exception of sacrificed animals. Stable carbon and nitrogen isotope data indicate that dogs were only buried where and when human diets were relatively rich in aquatic foods, which here most likely included river and lake fish and Baikal seal (Phoca sibirica). Generally, human and dog diets appear to have been similar across the study subregions, and this is important for interpreting their radiocarbon dates, and comparing them to those obtained on the region's human remains, both of which likely carry a freshwater old carbon bias. Slight offsets were observed in the isotope values of dogs and humans in our samples, particularly where both have diets rich in aquatic fauna. This may result from dietary differences between people and their dogs, perhaps due to consuming fish of different sizes, or even different tissues from the same aquatic fauna. This paper also provides a first glimpse of the DNA of ancient canids in Northeast Asia.

Infant and child diet in Neolithic hunter-fisher-gatherers from Cis-Baikal, Siberia: intra-long bone stable nitrogen and carbon isotope ratios.

Analysis of stable nitrogen and carbon isotopes (δ(15) N and δ(13) C) from subadults and adults allows for assessment of age-related dietary changes, including breastfeeding and weaning, and adoption of an adult diet. In one of the first studies of hunter-fisher-gatherer subadults from Eurasia, three Neolithic (8,800-5,200 calBP) mortuary sites from southwestern Siberia are analyzed to evaluate hypothesized differences in weaning age between Early versus Late Neolithic groups. An intra-individual sampling methodology is used to compare bone formed at different ages. Collagen samples (n = 143) from three different growth areas of long bones-the proximal metaphysis, diaphysis, and distal metaphysis-were obtained from 49 subadults aged birth to 10 years. In infants (birth to 3 years, n = 23) contrasting the δ(15) N values of the metaphysis, which contains newer bone, to the δ(15) N values of the diaphysis, which contains older bone, permits a more precise determination of breastfeeding-weaning status. In Early and Late Neolithic groups breast milk was the major protein source until the age of 2-3 years. However, there are differences in the age of weaning completion and duration: Early Neolithic groups weaned their infants at a later age and over a shorter amount of time. Differences may have affected infant morbidity and mortality, and female fecundity and inter-birth intervals. Stable isotope values in older subadults (4-10 years, n = 26) do not differ from adults suggesting the absence of age-based food allocation.

Activity-induced dental modification in holocene siberian hunter-fisher-gatherers.

The use of teeth as tools provides clues to past subsistence patterns and cultural practices. Five Holocene period hunter-fisher-gatherer mortuary sites from the south-western region of Lake Baikal, Siberia, Russian Federation, are observed for activity-induced dental modification (AIDM) to further characterize their adaptive regimes. Grooves on the occlusal surfaces of teeth are observed in 25 out of 123 individuals (20.3%) and were most likely produced during the processing of fibers from plants and animals, for making items such as nets and cordage. Regional variation in the frequency of individuals with occlusal grooves is found in riverine versus lakeshore sites. This variation suggests that production of material culture items differed, perhaps in relation to different fishing practices. There is also variation in the distribution of grooves by sex: grooves are found predominately in females, except at the Late Neolithic-Bronze Age river site of Ust'-Ida I where grooves are found exclusively in males. Occlusal grooves were cast using polyvinylsiloxane and maxillary canine impressions were examined by scanning electron microscopy (SEM) to determine striation patterns. Variation in striae orientation suggests that a variety of activities, and/or different manufacturing techniques, were involved in groove production. Overall, the variability in occlusal groove frequency, sex and regional distribution, and microscopic striae patterns, points to the multiplicity of activities and ways in which people used their mouths and teeth in cultural activities.

Comparison of microcomputed tomographic and microradiographic measurements of cortical bone porosity.

Cortical bone is perforated by a network of canals that have a significant impact upon its material properties. Microcomputed tomography offers the possibility of noninvasively visualizing and quantifying cortical pores in both two and three dimensions. Establishing how two-dimensional (2D) microcomputed tomographic (microCT) analysis compares with conventional methods for analyzing cortical porosity is an important prerequisite for the wider adoption of this technique and the development of three-dimensional (3D) analysis. Therefore, we compared porosity-related parameters from 2D microcomputed tomographic images with those from matching microradiographic sections. Samples from five human femora were scanned at a 10-microm resolution and then sequentially sectioned and microradiographed. An average of eight image pairs were produced from each femur (total, n = 41). The repeatability and comparability of the two techniques was assessed for three parameters; cortical porosity (%), mean pore area (microm(2)), and pore density (pores/mm(2)). For repeatability, no significant difference ( P > 0.05) was found between the two methods for cortical porosity and mean pore area; however, pore density differed significantly ( P < 0.001). For comparability, the bias (+/- error) between the methods was found to be 0.51% (+/-0.31%) for cortical porosity and -155 microm(2) (+/-293 microm(2)) for mean pore area. The bias for pore density was dependent upon measurement size with microcomputed tomographic images having 14% (+/-9.3%) fewer pores per millimeter squared. The qualitative and quantitative similarities between the two techniques demonstrated the utility of 2D microcomputed tomographic for cortical porosity analysis. However, the relatively poor results for pore density revealed that a higher resolution (<10 microm) is needed to consistently visualize all cortical pores in human bone.

Automated quantitation of hemoglobin-based blood substitutes in whole blood samples.

It is necessary to develop methods for accurate monitoring of cell-free hemoglobin in circulation. Routine monitoring of circulating cell-free hemoglobin will be useful for evaluating the efficacy of blood substitute administration andfor determining the clearance rates of the blood substitute from circulation. In addition, discriminating between cell-free hemoglobin and cell-associated hemoglobin will enable accurate determination of RBC indices, mean cell hemoglobin and mean corpuscular hemoglobin concentration, in individuals receiving hemoglobin-based blood substitutes. As colorimetric methods used by hematology analyzers to quantitate the hemoglobin value of a blood sample cannot distinguish between cell-associated and cell-free hemoglobin, it is currently not feasible to quantitate the levels of hemoglobin substitutes in circulation. The advent of a technology that measures volume and hemoglobin concentration of individual RBCs provides an alternative strategy for quantitating the cell-associated hemoglobin in a blood sample. We document that the combined use of cell-based and colorimetric hemoglobin measurements provides accurate discrimination between cell-associated and cell-free hemoglobin over a wide range of hemoglobin levels. This strategy should enable rapid and accurate monitoring of the levels of cell-free hemoglobin substitutes in the circulation of recipients of these blood substitutes.

Investigating the weaning process in past populations.

The 19th century St. Thomas'Anglican churchyard in Belleville, Ontario, Canada is associated with a large and well-preserved infant skeletal collection (n = 149) and good-quality parish records that document interments in the graveyard (1821-1874). By using a combination of historical demographic and stable nitrogen isotope analyses on the parish records and skeletal remains, respectively, a general pattern of extended nursing for about 14 months, introduction of foods other than breast milk by around 5 months of age, and variation in breast-feeding and weaning behaviours were detected for St. Thomas' infants. The results demonstrate that it is possible to go beyond the concept of weaning age to explore the weaning process in past populations when appropriate and large samples of documentary and skeletal data are available.

Dental caries in nineteenth century upper Canada.

This study examines the presence of dental caries in a large sample of adult skeletons from the 19th century cemetery of St. Thomas' Anglican Church in Belleville, Ontario. The cemetery was used from 1821 to 1874. Caries prevalence and frequencies of diseased and missing teeth were calculated both by observing summary statistics of individual rates and by the total sample of teeth. Postmortem tooth loss is low in this sample and antemortem tooth loss is highest in first mandibular molars, all other molars and then premolars. Age at death, but not sex, was found to be significantly related to the overall Caries Rate while both age and sex were significantly associated with the Diseased-Missing Index. The increase in diseased and missing teeth in older individuals is expected while the sex difference is not explained by simple dietary factors. When compared to reports on British and American samples, caries and antemortem tooth loss in the St. Thomas' sample is most similar to a pre-1850 British group and higher than American samples. Although there is undoubtedly a complex of factors contributing to caries prevalence in this sample, more data are required from large historic samples, particularly from the American northeast and late 19th century Britain, to have a clearer understanding of the influence of diet, cultural, and environmental factors affecting caries rates in historic populations.

Status and diet in precontact highland Ecuador.

Excavation at the Ecuadorian highland site of La Florida in suburban Quito revealed six deep shaft tombs yielding high-status individuals (n = 9) as well as apparent sacrifices and other low-status individuals (n = 23). Determination of sex and age at death of the recovered skeletal remains resulted in a sample of 32 individuals aged from approximately 7 to 50 years of age. The sample of 18 individuals over the age of 18 years included 14 females and 4 males. Temporally, the remains are assigned to the Chaupicruz Phase (circa 100 to 450 AD) of the Regional Developmental Period. In this study, we analyze stable isotopes of carbon and nitrogen from human bone in order to compare the diets of the high- and low-status individuals. Stable carbon isotope analyses were carried out on preserved protein and biological apatite (bioapatite), and stable nitrogen isotope analyses were carried out on preserved protein. There is a statistically significant difference in delta 13C between the two groups for both protein and mineral sources of carbon with evidence for the greater consumption of maize in the high-status group. There is no significant difference in delta 15N between the two groups, nor is there a significant difference in the spacing between protein and mineral delta 13C values between the two groups. Ethnohistorical evidence for the 16th century AD provides the expectation that the only dietary difference was the higher consumption of animal protein by the elite. There is no evidence for this based on the bone chemistry data from La Florida.(ABSTRACT TRUNCATED AT 250 WORDS)

Age differences in stable carbon and nitrogen isotope ratios in a population of prehistoric maize horticulturists.

Stable carbon isotope ratios in prehistoric human bone collagen have been used extensively to document the introduction and intensification of maize horticulture in notheastern North America. Most previous studies are based on small samples of adults who are assumed to characterize the diet of the population. In this study, all 29 individuals buried within an Ontario Iroquoian village site dated A.D. 1530-1580 were analysed for stable isotopes of carbon and nitrogen. Age distribution of the sample ranges from preterm to elderly. Significant negative correlations between age and delta 13C, and age and delta 15N values were found. High delta 13C values in infants and young children (delta 13C = -6.8 to -12.3) suggest a weaning diet high in maize. High delta 15N values in infants relative to adults suggest a trophic level effect during breast-feeding which has been reported in a modern sample by Tuross et al. (Am. J. Phys. Anthropol., 1993). In addition to the isotopic evidence for extremely high carbohydrate (maize) intake, the MacPherson sample includes two juveniles aged 3-4 years, exhibiting circular caries. No other cases of this condition are known in the extensively studied southern Ontario skeletal collections. Together the evidence from dentition and stable carbon isotopes indicates a very high carbohydrate diet in subadults. Circular caries result from developmental stress during enamel formation with subsequent caries formation in areas of thinner enamel. These findings are relevant to studies of infant and early childhood morbidity and mortality among prehistoric maize horticulturists.

Oxidative phosphorylation system during steady-state hypoxia in the dog brain.

The relationship between biochemical and physiological responses and tissue O2 during hypoxia was investigated in vivo in the dog brain by 31P nuclear magnetic resonance (NMR) spectroscopy. Our findings demonstrate how ATP synthesis in the brain can be maintained during hypoxia because of compensatory changes in NADH, ADP, and Pi. Eleven beagle dogs were anesthetized and mechanically ventilated, and a steady-state graded hypoxia was induced by decreasing the fraction of inspired O2 (FIO2) stepwise at 20-min intervals. Biochemical metabolites were measured using 31P-NMR and fluorescence spectroscopy. When sagittal sinus O2 partial pressure (PVO2) had decreased to 15 Torr, NADH increased by 30%, Pi increased by 50%, and phosphocreatine (PCr) decreased by 20%. In contrast, ATP remained constant. There was a 10% increase in ADP in dogs that maintained a steady temperature, but ADP decreased by as much as 30% in dogs in which body temperature decreased with the falling PVO2. PCr/Pi was logarithmically related to the phosphorylation potential during steady-state hypoxia. Compensation for the O2 lack is attributed to increases in ADP, Pi, and NADH as a result of the reciprocal relationship of the Michaelis-Menten equation. If the Michaelis-Menten constants (Km) of ADP, Pi, and O2 are the same as determined in vitro in mitochondria, the minimum brain cytosolic O2 capable of maintaining a steady-state ATP is near its Km (0.1 Torr) at a PVO2 of 7.5 Torr. At this critical O2 level, PCr/Pi is 0.9, intracellular pH is 6.75, phosphorylation potential is 38.5 mM-1, and the calculated maximum velocity of ATP formation by oxidative phosphorylation is 55% of normal.