Yuichi I. Naito

Quantitative Evaluation of Marine Protein Contribution in Ancient Diets Based on Nitrogen Isotope Ratios of Individual Amino Acids in Bone Collagen: An Investigation at the Kitakogane Jomon Site

Nitrogen stable isotopes analysis of individual bone collagen amino acids was applied to archeological samples as a new tool for assessing the composition of ancient human diets and calibrating radiocarbon dates. We used this technique to investigate human and faunal samples from the Kitakogane shell midden in Hokkaido, Japan (5,300-6,000 cal BP). Using compound-specific nitrogen isotope analysis of individual amino acids, we aimed to estimate i) the quantitative contribution of marine and terrestrial protein to the human diet, and ii) the mean trophic level (TL) from which dietary protein was derived from marine ecosystems. Data were interpreted with reference to the amino acid trophic level (TL(AA)) model, which uses empirical amino acid delta(15)N from modern marine fauna to construct mathematical equations that predict the trophic position of organisms. The TL(AA) model produced realistic TL estimates for the Kitakogane marine animals. However, this model was not appropriate for the interpretation of human amino acid delta(15)N, as dietary protein is derived from both marine and terrestrial environments. Hence, we developed a series of relevant equations that considered the consumption of dietary resources from both ecosystems. Using these equations, the mean percentage of marine protein in the Kitakogane human diet was estimated to be 74%. Although this study is one of the first systematic investigations of amino acid delta(15)N in archeological bone collagen, we believe that this technique is extremely useful for TL reconstruction, palaeodietary interpretation, and the correction of marine reservoir effects for radiocarbon dating.

Ecological niche of Neanderthals from Spy Cave revealed by nitrogen isotopes of individual amino acids in collagen

This study provides a refined view on the diet and ecological niche of Neanderthals. The traditional view is that Neanderthals obtained most of their dietary protein from terrestrial animals, especially from large herbivores that roamed the open landscapes. Evidence based on the conventional carbon and nitrogen isotopic composition of bulk collagen has supported this view, although recent findings based on plant remains in the tooth calculus, microwear analyses, and small game and marine animal remains from archaeological sites have raised some questions regarding this assumption. However, the lack of a protein source other than meat in the Neanderthal diet may be due to methodological difficulties in defining the isotopic composition of plants. Based on the nitrogen isotopic composition of glutamic acid and phenylalanine in collagen for Neanderthals from Spy Cave (Belgium), we show that i) there was an inter-individual dietary heterogeneity even within one archaeological site that has not been evident in bulk collagen isotopic compositions, ii) they occupied an ecological niche different from those of hyenas, and iii) they could rely on plants for up to ∼20% of their protein source. These results are consistent with the evidence found of plant consumption by the Spy Neanderthals, suggesting a broader subsistence strategy than previously considered.

Dietary Reconstruction of the Okhotsk Culture of Hokkaido, Japan, Based on Nitrogen Composition of Amino Acids: Implications for Correction of 14C Marine Reservoir Effects on Human Bones

The relative contribution of marine-derived carbon in the ancient diet is essential for correcting the marine reservoir effect on the radiocarbon age of archaeological human remains. In this study, we evaluated the marine protein consumption of 3 human populations from the Okhotsk culture (about AD 550-1200) in Hokkaido, Japan, based on stable carbon and nitrogen isotopic compositions in bulk bone collagen as well as the nitrogen isotopic composition of glutamic acid and phenylalanine. Despite the similarity of carbon and nitrogen isotopic composition of bulk collagens, nitrogen isotopic composition of their constituent amino acids suggests differences in fur seal contributions among northern Hokkaido (0-24% for Kafukai 1, 0-10% for Hamanaka 2) and eastern Hokkaido (78-80% for Moyoro) populations. It suggests that nitrogen composition of glutamic acid and phenylalanine could provide a detailed picture of ancient human subsistence.

Association of protein intakes and variation of diet-scalp hair nitrogen isotopic discrimination factor in Papua New Guinea highlanders.

OBJECTIVES We present new nitrogen isotopic discrimination factor between diets and scalp hairs (Δ(15) NHair-Diet : δ(15) NHair - δ(15) NDiet ) for indigenous residents in three communities in the Papua New Guinea Highlands who consumed various amounts and qualities of protein. The Δ(15) N is important for precise evaluation of the dietary habits of human populations; in both contemporary and traditional lifestyles. Several hypotheses have been proposed regarding factors that affect Δ(15) N values, based largely on observations from animal feeding experiments. However, variations and factors controlling Δ(15) N in humans are not well understood, mainly due to the difficulty of controlling the diets of participants. MATERIALS AND METHODS These residents were studied because they have maintained relatively traditional dietary habits, which allow quantitative recording of diets. Δ(15) N was estimated by comparing hair δ(15) N values to mean dietary δ(15) N values calculated from the recorded intake of each food item and their δ(15) N values. RESULTS The results showed that: i) there was a significant difference in Δ(15) N among study locations (3.9 ± 0.9‰ for most urbanized, 5.2 ± 1.0‰ for medium and 5.0 ± 0.9‰ for least urbanized communities; range = 1.2-7.3‰ for all participants); and ii) estimated Δ(15) N values were negatively correlated with several indicators of animal protein intake (% nitrogen in diet: range = 0.9-7.6%). DISCUSSION We hypothesize that a combination of several factors, which presumably included urea recycling and amino acid and protein recycling and/or de novo synthesis during metabolic processes, altered the Δ(15) N values of the participants.

Characterization of the gut microbiota of Papua New Guineans using reverse transcription quantitative PCR.

There has been considerable interest in composition of gut microbiota in recent years, leading to a better understanding of the role the gut microbiota plays in health and disease. Most studies have been limited in their geographical and socioeconomic diversity to high-income settings, and have been conducted using small sample sizes. To date, few analyses have been conducted in low-income settings, where a better understanding of the gut microbiome could lead to the greatest return in terms of health benefits. Here, we have used quantitative real-time polymerase chain reaction targeting dominant and sub-dominant groups of microorganisms associated with human gut microbiome in 115 people living a subsistence lifestyle in rural areas of Papua New Guinea. Quantification of Clostridium coccoides group, C. leptum subgroup, C. perfringens, Bacteroides fragilis group, Bifidobacterium, Atopobium cluster, Prevotella, Enterobacteriaceae, Enterococcus, Staphylococcus, and Lactobacillus spp. was conducted. Principle coordinates analysis (PCoA) revealed two dimensions with Prevotella, clostridia, Atopobium, Enterobacteriaceae, Enterococcus and Staphylococcus grouping in one dimension, while B. fragilis, Bifidobacterium and Lactobacillus grouping in the second dimension. Highland people had higher numbers of most groups of bacteria detected, and this is likely a key factor for the differences revealed by PCoA between highland and lowland study participants. Age and sex were not major determinants in microbial population composition. The study demonstrates a gut microbial composition with some similarities to those observed in other low-income settings where traditional diets are consumed, which have previously been suggested to favor energy extraction from a carbohydrate rich diet.

Association between sex inequality in animal protein intake and economic development in the Papua New Guinea highlands: The carbon and nitrogen isotopic composition of scalp hair and fingernail.

OBJECTIVES People in the Papua New Guinea Highlands consume sweet potatoes as their dietary staple; consumption of animal protein is limited. In such societies with marginal protein intake, the intra-household allocation of animal protein in terms of sex or age is of importance. The objective of this study was to investigate how the allocation pattern of protein-rich foods by sex and age is associated with economic development in the Papua New Guinea Highlands. METHODS The carbon and nitrogen isotopic compositions of fingernails, collected in 1995 in two areas (Tari and Port Moresby [the national capital where Tari migrants resided]), and of scalp hair, collected in 2007, 2012, and 2013 in three areas of different degree of economic development (Levani, Tari, and Goroka) were analyzed. RESULTS Analysis of fingernail samples showed that δ(15)N was lower in rural communities than in the urban migrant community, while a sex difference in δ(15)N (higher in males than in females) was found in the former but not in the latter community. Age was not associated with either δ(15)N or δ(13)C values. The analysis of scalp hair samples showed that δ(15)N values were lowest in Levani, the least developed area. Furthermore, there were statistically significant sex differences in δ(15)N values in Levani but not in Tari and Goroka. Age was not associated with either δ(15)N or δ(13)C values. DISCUSSION The sex inequality in animal protein consumption seems to have decreased as the communities in the Papua New Guinea Highlands have experienced economic development.

Isotopic analyses suggest mammoth and plant in the diet of the oldest anatomically modern humans from far southeast Europe

Relatively high 15N abundances in bone collagen of early anatomically modern humans in Europe have often been interpreted as a specific consumption of freshwater resources, even if mammoth is an alternative high 15N prey. At Buran-Kaya III, access to associated fauna in a secured archaeological context and application of recently developed isotopic analyses of individuals amino acids offer the opportunity to further examine this hypothesis. The site of Buran-Kaya III is located in south Crimea and has provided a rich archaeological sequence including two Upper Palaeolithic layers, from which human fossils were retrieved and directly dated as from 37.8 to 33.1 ka cal BP. Results from bulk collagen of three human remains suggests the consumption of a high 15N prey besides the contribution of saiga, red deer, horse and hare, whose butchered remains were present at the site. In contrast to bulk collagen, phenylalanine and glutamic acid 15N abundances reflect not only animal but also plant protein contributions to omnivorous diet, and allow disentangling aquatic from terrestrial resource consumption. The inferred human trophic position values point to terrestrial-based diet, meaning a significant contribution of mammoth meat, in addition to a clear intake of plant protein.

A high burden of asymptomatic gastrointestinal infections in traditional communities in Papua New Guinea

Abstract. Stool samples were collected from 148 healthy adults living a traditional subsistence lifestyle in Papua New Guinea and screened for enteric pathogens using real-time RT-PCR/PCR assays. Enteric pathogens were detected in a high proportion (41%) of individuals. Clear differences were observed in the detection of pathogens between highland and lowland communities. In particular, there was a marked difference in detection rates of norovirus GII (20% and 0%, respectively) and Shigella sp. (15% and 0%, respectively). Analysis of the relationship between enteric pathogen carriage and microbial community composition of participants, using box plots to compare specific normal flora population numbers, did not suggest that gut microbial composition was directly associated with pathogen carriage. This study suggests that enteric pathogens are common in healthy individuals in Papua New Guinean highland communities, presumably acting as a reservoir of infection and thus contributing to a high burden of gastrointestinal illnesses.