Evidence for chronic omega-3 fatty acids and ascorbic acid deficiency in Palaeolithic hominins in Europe at the emergence of cannibalism

Evidence for chronic omega-3 fatty acids and ascorbic acid deficiency in Palaeolithic hominins in Europe at the emergence of cannibalism

Quaternary Science Reviews 157 (2017) 176e187 Contents lists available at ScienceDirect Quaternary Science Reviews journal homepage: www.elsevier.co...

1MB Sizes 10 Downloads 65 Views

Quaternary Science Reviews 157 (2017) 176e187

Contents lists available at ScienceDirect

Quaternary Science Reviews journal homepage: www.elsevier.com/locate/quascirev

Evidence for chronic omega-3 fatty acids and ascorbic acid deficiency in Palaeolithic hominins in Europe at the emergence of cannibalism J.L. Guil-Guerrero Food Technology Division, CeiA3, University of Almería, 04120, Almería, Spain

a r t i c l e i n f o

a b s t r a c t

Article history: Received 15 April 2016 Received in revised form 16 December 2016 Accepted 19 December 2016 Available online 10 January 2017

At the Middle-Upper Palaeolithic (M/UP) transition in Western Europe, hominins depended mostly on terrestrial mammals for subsistence, being pointed out that reliance on reindeer (Rangifer tarandus) would have promoted declines in human population densities during that period. Food-composition tables have been compiled for hominins at the M/UP transition, listing protein, fat, energy, different omega-3 fatty acids and ascorbic acid concentrations. These data were used to compute the regular relations between fatty and lean tissues of the main hunted food-animals to meet hominin energy needs. Then, with daily protein intake considered critical, the optimal contribution of the different omega-3 fatty acids from different hunted species to hominin diets were computed. Several faunal assemblages from different human sites at different M/UP periods were used to assess the overall daily intake of the various omega-3 fatty acid classes. The results of the calculations made in this work are quite clear; hominins at the M/UP transition had a deficit of both omega-3 fatty acids and ascorbic acid. Data on human organs summarized here are also conclusive: these contain such nutrients in amounts much higher than reached in the corresponding mammal organs consumed, and thus could have been alternative sources of those nutrients for Palaeolithic hominins. Therefore, nutritional cannibalism detected at such times could have had the function of alleviating these deficits. The evolutionary advantages gained by the consumption of the various omega-3 fatty acids of human origin are also discussed. © 2016 Elsevier Ltd. All rights reserved.

Keywords: DHA EPAþDHA Omega-3 PUFA n-3 PUFA Cannibalism Middle-Upper Palaeolithic transition Reindeer

1. Introduction Assessing the nutritional status during the Palaeolithic is of great interest because detecting diseases associated with different diets in the past offers an understanding and improvement of the nutrition of current humans. The term “essential nutrients” refers to those that the human body must derive from foods (Hockett and Haws, 2003). Today, many studies examine the availability of essential fatty acids (EFAs) for humans in the Palaeolithic because, among other reasons, anthropological and epidemiological works have indicated that during the Palaeolithic the human diet had an omega-6 (n-6) to omega-3 (n-3) EFA ratio ~1, while today in Western diets the ratio is ~16:1, which may be responsible for the pathogenesis of many diseases (Simopoulos, 2006). Conversely, studies on the occurrence and availability of ascorbic acid in the Palaeolithic are scarce. FAs, the fat components in addition to glycerol, can be classified

E-mail address: [email protected] http://dx.doi.org/10.1016/j.quascirev.2016.12.016 0277-3791/© 2016 Elsevier Ltd. All rights reserved.

into two main types: saturated and unsaturated, depending on whether or not they have double bonds in their structure. Monounsaturated fats have one double bond, while polyunsaturated fats have more than one double bond. For the latter, the n-3- omegareference system indicates the number of carbons, the number of double bonds, and the position of the double bond closest to the omega carbon, counting from the omega carbon (which is numbered 1 for this purpose). Monounsaturated FAs (MUFAs) are mainly n-9 FAs, for instance oleic acid (OA, 18:1n-9), while polyunsaturated FAs (PUFAs) belong to two families: n-6 and n-3 (Fig. 1). Two C18 PUFA, linoleic acid (LA, 18:2n-6) and a-linolenic acid (ALA, 18:3n-3), are considered EFAs because they cannot be synthesised by humans, and therefore they must be included in the diet. Conversely, the C20-22 very-long-chain PUFAs (VLCPUFAs) can be biosynthesised by the consecutive action of several enzymes from their respective dietary EFA precursors: arachidonic acid (ARA, 20:4n-6) from LA, and eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) from ALA (GuilGuerrero, 2007). Used as an energy source, n-6 and n-3 PUFA also influence cell

J.L. Guil-Guerrero / Quaternary Science Reviews 157 (2017) 176e187

Abbreviations ALA a-linolenic acid (18:3n-3) AFFSA French Food Safety Agency ARA arachidonic acid (20:4n-6) DHA docosahexaenoic acid (22:6n-3) DPA docosapentaenoic acid (22:5n-3) EFA essential FA EFSA European Food Safety Authority EPA eicosapentaenoic acid (20:5n-3) FA fatty acid LA linoleic acid (18:2n-6) LCPUFA long-chain PUFA M/UP Middle- Upper Palaeolithic NISP number of identified specimens MNI minimum number of individuals MUFA monounsaturated FA OA oleic acid (18:1n-9) PUFA polyunsaturated FA S/DF subcutaneous and depot fats (fatty tissues) VLCPUFA very-LCPUFA

and tissue metabolism, function, and responsiveness to hormonal and other signals (Calder, 2015). Intracellular activities of FAs include: the regulation of membrane structure and function (Calder, 2012); regulation of intracellular signalling pathways, transcription factor activity, and gene expression (Georgiadi and Kersten, 2012); and regulation of the production of bioactive lipid mediators (Calviello et al., 2013). Such activities depend on AA, EPA, and DHA. These have important roles in immune regulation and inflammation (Miles and Calder, 2012). Through these effects, VLCPUFA influence cardiovascular diseases and a wide range of other illnesses, such as metabolic diseases (e.g. type 2 diabetes), inflammatory disorders, and cancer (Calder, 2012). Moreover, VLCPUFA are involved in the healthy performance of several physiological functions, for instance blood pressure and blood clotting, and the proper development and functioning of the brain and nervous system. A crucial fact is that DHA is scanty outside the central nervous system, representing a high proportion only in the lipids in the retina and grey matter of the brain (Guil-Guerrero, 2007; Wainwright, 2002). Today, ample evidence indicates that the bioconversion of ALA to EPA and DHA is very low, although the exact extent is subject to controversy, being the capacity to generate DHA from ALA higher in women than in men. Based on previous studies, Harris (2012) estimated an average bioconversion of ALA to EPA and DHA of 3.7 and 0.8%, respectively, whereas for n-6 PUFA-enriched diets the bioconversion is reduced by 40e50% (Guil-Guerrero, 2007). Such conversions are rate-limited by hepatic D6-desaturase, which rapidly declines with age (Bradbury, 2011). In men, this decline has been demonstrated to be between three-to six-fold lower in older than in younger individuals (Burdge et al., 2003). Thus, the maintenance of the EPA and DHA status in older individuals may depend primarily upon dietary intakes of preformed EPA and DHA (Burdge and Calder, 2005). On the other hand, for women, some differences have been reported in the activity of this enzyme among ethnic groups (Gray et al., 2013). Due to low efficiency in the conversion in the n-3 route, it is recommended that EPA and DHA be derived from additional sources, and thus they are considered “conditionally essential” nutrients (Bradbury, 2011). All mammalian species have high DHA and ARA levels in their


brain (Crawford and Sinclair, 1976) and thus their intake restriction will determine limitations to brain development. Such a restriction could have dramatic consequences for Homo spp., because of the large brain size, in contrast to previous hominins, such as Australopithecus spp., which had a low encephalization rate (Crawford et al., 1999). In contrast with n-3 EFAs, the significance of ascorbic acid (Vitamin C) in Palaeolithic diets has hardly been studied, although there are indications for consumption deficiency, which could have led to significant pathologies (Ortner and Ericksen, 1997). Ascorbic acid is an essential nutrient for humans, participating in collagen synthesis, and has antioxidant functions as a scavenger of free radicals. It also seems to protect tissues from harmful oxidative products and maintains certain enzymes in their required reduced forms (Padh, 1990). Vitamin C deficiency shows symptoms such as bruising, arthralgias, or joint swelling, and common signs are pedal oedema, bruising, or mucosal changes, myalgias, and general fatigue (Olmedo et al., 2006). The M/UP transition is a period that refers to the age of the most recent Neanderthal fossils and the earliest modern human remains in Europe, and the inferred overlap between the Ch^ atelperronian and the Aurignacian (Bocherens et al., 2014). This period is vital to understand diverse constraints on the current human society, from religious worship and artistic expression to administrative structures (Roebroeks, 2008). In Western Europe, the M/UP transition included a widespread adoption of blade-based toolkits, the emergence of artistic behaviour, and a shift towards modern human anatomical features. Data from this period reveal a significant climatic deterioration, which was associated with a reduction in mammalian species diversity (Morin, 2008). Today there is clear evidence for the strong dependence on animal foods by M/UP hominins in Europe. Both lithic and faunal assemblages and stable isotope analysis provides a direct measure of human diets in the past and supports the hunting hypothesis for M/UP populations  et al., 2009; Craig (Richards et al., 2000, 2000b, 2008; Garcia-Guixe et al., 2010; Drucker and Henry-Gambier, 2005; Bocherens et al., 2005; Grayson and Delpech, 2002; Wißing et al., 2015). The isotopic evidence indicates that this scenario is true in all cases for Neanderthals, which were top-level carnivores whereas, by contrast, early modern humans (z40,000 to z27,000 y BP) showed a wider range of d15N values, and evidence from some individuals point to the consumption of aquatic (marine and freshwater) resources (Richards and Trinkaus, 2009). However, a shift in d15N at the base of the terrestrial foodweb could have been responsible for such a pattern, with a preserved foodweb structure without any significant change in the diet composition before and after the M/UP transition (Bocherens et al., 2014). At the M/UP transition, when subsistence in Europe heavily depended on animal foods, the question arises as to whether animal-food resources could provide those hunters certain essential nutrients necessary to maintain a good health status, and whether M/UP hominins could have exploited n-3 PUFA and ascorbic acid resources in addition to those from hunting. This study attempts to resolve these issues. 2. Material and methods The relevant literature was searched for articles dealing with FAs and ascorbic acid composition of mammals living at the M/UP transition, as well as other articles on the content of these nutrients in human organs. Also, the literature on M/UP assemblages was scrutinized, and articles about the emergence of cannibalism around the world, as well as others concerning Inuit nutrition. Data were acquired from databases such as Google Scholar, Scopus, and other similar resources.


J.L. Guil-Guerrero / Quaternary Science Reviews 157 (2017) 176e187

Fig. 1. LA and ALA, the parent compounds of the omega-6 (n-6) and omega-3 (n-3) PUFA families, are obtained primarily from vegetable oils, but are also found in foods of plant and animal origin. They give rise to longer chain derivatives inside the body. Due to low efficiency of conversion of ALA to SDA and further n-3 VLCPUFA, EPA and DHA, it is recommended to obtain EPA and DHA from additional sources.

The minimum daily protein to meet humans need was calculated according to Paddon-Jones and Rasmussen (2009) at 0.8 g/ kg$day, while the maximum safe protein daily intake was taken from Bilsborough and Mann (2006), which was 2.5 g/kg$day. The estimated body weight for M/UP humans has been estimated as the average values given for Medium Palaeolithic individuals, both males and females, of 65 kg (Hermanussen, 2003), which closely agrees with calculations from Mathers and Henneberg (1995). The total energy expenditure for hominins was computed for both male and female Neanderthals, through the following values: 4000e7000 kcal/day in males and 3000e5000 kcal/day in females (Snodgrass and Leonard, 2009); 3000e5000 kcal/day in females and 4000e6000 kcal/day in males (Sorensen and Leonard, 2001); and 3360e4480 kcal/day for males (Steegmann et al., 2002), the mean of these values being ~4500 kcal/day. Values for n-3 PUFAs, protein, and fat of free-range mammals commonly consumed by Palaeolithic people in Europe are summarized in Table S1 (meat), and Table S2 (viscera). Some other animals occasionally eaten, such as are carnivores, hares, birds, and reptiles, have not been considered, because of their low incidence of appearance in most Palaeolithic assemblages. Given an absence of values for some nutrients of subcutaneous and depot fats (S/DF, the fats that are stored is some organs mainly as an energy

reservoir), average values for protein, fat, and energy of such organs have been estimated through data from USDA Nutrient Data Base for separable and raw fat from pork, beef, lamb, and veal; and for raw subcutaneous fat from lamb and veal (Table S3). Similar data for viscera are summarized in Table S4. The data for n-3 PUFA profiles of human organs are summarized in Table S5. For faunal assemblages, the minimum number of individuals (MNI) was used for dating, as well as the number of identified specimens (NISP), in order to assess the supply of the various n-3 PUFA to the hominin diet. This was done by calculating the total carcass weight for each species using published data and then the relative proportion of each species to the total live weight. Thereafter, the n-3 PUFA data for meat and S/DF of each species were used to calculate the total supply of the various n-3 PUFA at the optimum proportion meat:S/DF for each species, and adding up the different n-3 PUFA computed for each of them. 3. Results and discussion 3.1. Potential foods at the M/UP transition Today, stable-isotope analyses have helped to reveal the nature of foods in Europe for M/UP hominins. Such studies provide

J.L. Guil-Guerrero / Quaternary Science Reviews 157 (2017) 176e187

objective data showing that the diets of hominins living in Europe during the Palaeolithic were indistinguishable from top trophic level carnivores such as arctic foxes and wolves. From another perspective, Kaplan and Hill (1992) computed proportions of plants and animal foods in hunter-gatherer diets, and found that some peoples living in regions close to the Arctic were strongly dependent until recently on animal foods, as in the case of Eskimos (Greenland) and Nunamiut (Alaska), which consume such foods at 96 and 99%, respectively, and it is believed that Palaeolithic populations, who lived in similar habitats, must also have depended heavily on food of animal origin. Considering both the locations where the European Palaeolithic human settlements are usually found as well as food remains found therein, plant foods, marine mammals, and lacustrine resources seem to have played a marginal role in the diet of these Palaeolithic human populations (Cordain et al., 2002; Morin, 2008). However, peripheral populations of Neanderthals and some others of anatomically modern humans (AMH) consumed variable amounts of plant foods and fish, as denoted by isotopic analyses (Richards et al., 2009). For instance, Neanderthals exploited lacustrine fish, starchy plants, and wood in southern France (Hardy and Moncel, 2011); molluscs in southern s-Sa nchez et al., 2011); some plant foods such as acorn Spain (Corte flour, Typha rhizome and emmer flour in the southern Italy and Czech Republic (Revedin et al., 2015), and some others. All these can be considered seasonal exceptions confirming a clear trend. Although such foods could have provided nutrients and energy, in most cases they could never have been considered noteworthy n-3 PUFA sources. Fish resources (Table S6) were not a major part of M/UP hominins diet, as demonstrated by isotopic values (Richards and Trinkaus, 2009), and their remains are scarce in most M/UP assemblages in Europe. Assuming that M/UP transition hominins had more or less similar behaviour to that of other present-day peoples living in comparable habitats, the role of fishing in their subsistence can be understood. For instance, fish are available to the Ojibwa people (South-eastern Canada and North-eastern North America) during all seasons, although most fishing occurs during the summer months, from June onwards, while in winter they spear trout and sturgeon through the ice (Russ, 2013). However, M/UP hominins are not very likely to have had the necessary technology to open holes through thick ice sheets for fishing, so this resource must have been exploited in such times necessarily in rivers and lakes during the short thaws of summer. In any case, the freshwater fish that appear in some Palaeolithic settlements could have supplied major amounts of EPA þ DHA to the hominins of those times, satisfying their daily necessities (Table S6); nonetheless, as has been argued elsewhere, fish consumption was seasonal and was never a substitute for hunting. From all potentially ingested plant foods, only a few seeds could have provided n-3 PUFAs in appropriate amounts to M/UP hominins (Table S7). These are seeds of Linaceae, Cruciferae, Rosaceae, Fabaceae and some others, in which ALA can reach considerable amounts. However, some of these seeds, for instance Cruciferae and Fabaceae, can reach high toxicity when ingested in moderate amounts (Dolan et al., 2010), and such resources could have had a minor role as n-3 PUFA suppliers, given that they lack C20-22 n-3 VLCPUFA (Guil-Guerrrero, 2014). Moreover, the cold climate at the Palaeolithic in Europe possibly shortened the harvesting period for plant foods to maybe a month or two a year. Thus, considering all the above factors, there were few possibilities to meet the n-3 PUFA needs for M/UP hominins through the intake of any vegetable organs. Taking all of the above into account, fish and plant foods resources will not be taken into account hereafter for calculations of the daily diet of M/UP hominins.


3.2. Fatty acid profiles of M/UP hunted mammals The n-3 PUFA profiles of intramuscular fat content of commonly hunted M/UP mammals are summarized in Table S1. Note the minimal differences in the n-3 PUFA profiles among the various species, the percentages of the different n-3 PUFA in this order: ALA > DPA > EPA > DHA, although the total concentration of muscle fat greatly varies; that is, wild boar reached the highest fat value, 3.1 g/100 g tissue, followed by reindeer, with 2.2 g/100 g. The significance of these profiles in hominin nutrition will be discussed below. The n-3 PUFA profiles for viscera of M/UP hunted mammals are summarized in Table S2. Given an absence of values for protein and fat in these organs, data for similar animal foods from USDA Nutrient Database were used for calculations (Tables S3 and S4). The brain, for all the mammals considered, was the best source of fat and VLCPUFA, especially DHA. Concerning S/DF, monogastric mammals registered the highest values for n-3 PUFAs, with horse holding the foremost position (10.4 g n-3 PUFAS/100 g FAs), and ALA in most cases being the sole contributor to this profile. n-3 VLCPUFA are usually absent in S/DF tissues, although bear contains minor amounts, because of its omnivory. Ruminants showed a wide variation in n-3 PUFA content, with bison at the top of the range and reindeer, goat and elk at the bottom. For the remaining organs, the n-3 PUFA profiles of liver proved very similar to that of meat, although DPA reached higher percentages than ALA, and contained slightly more fat than did the muscles. On the other hand, the n-3 PUFA profiles of bone marrow were very similar to those of S/DF. 3.3. The supply of n-3 PUFA by M/UP mammals A diet based on animal foods would have provided undoubted benefits to M/UP hominins because such foods usually provide a higher ratio of energy gain to expenditure than do plant/fish-based foods (Simms, 1987; Hawkes and O'Connell, 1985). Nevertheless, this diet may also have caused disadvantages, as indicated by Rudman et al. (1973), who showed that the mean maximal rate of urea synthesis (MRUS) in normal subjects is 65 mg N/h per kg body wt0.75 and that protein intake that exceeded the MRUS resulted in hyperammonemia and hyperaminoacidemia. Accordingly, the maximum daily protein intake has been estimated as 2.5 g/kg$day (Bilsborough and Mann, 2006), and to circumvent the excess of dietary protein the preferred solution by most worldwide huntergatherers was a relative increase in total dietary fat from animal foods (Pryor, 2008). In this context, all the edible carcasses were processed to obtain fat (Speth and Spielmann, 1983). Anthropological and ethnographic data indicate that Stone Age humans consumed not just muscle tissue, but relished certain fatty portions of the carcass including the brain and marrow (Stiner, 1991; Silberbauer, 1981). However, the relative amount of S/DF and meat that a single prey provides is variable. For instance, Reimers (1984) indicated that S/DF in reindeer was approximately 1/3 of meat. Thus, it is likely that M/UP hominins had to hunt several times before their meat reserves were exhausted, in order to meet their daily energy needs. Another possibility is that they scavenged mammalian carcasses, which usually appear devoid of organs and flesh but still have enough fat adhering to the skin, able to be removed by using stone tools; thus, the scavenging behaviour would have been useful to complete the diet of M/UP hominins (Binford, 1984). Moreover, S/DF included in carcasses could also have been a suitable way to store food, since its stability against spoilage is higher than that of meat or any other viscera, due to the low water content. A notable n-3 PUFA-enriched organ is the brain. For a typical M/ UP mammal, it provides ~720 mg DHA and ~123 kcal by 100 g tissue


J.L. Guil-Guerrero / Quaternary Science Reviews 157 (2017) 176e187

(Table S2). However, its small size, z100e400 g for M/UP mammals (Crile and Quiring, 1940), and its low caloric content as well as its high degradability surely make it an erratic n-3 PUFA source, and this might be better regarded as a kind of ephemeral delicacy to be eaten by privileged members of the group, such as children, pregnant women, or aged individuals (Guil-Guerrero et al., 2013). In any case, it seems unrealistic to consider that the brain from any hunted mammal could be equitably divided among the members of a Palaeolithic hunter group. Probably, all the internal organs, such as brain, liver, heart, kidneys, spleen, testicles, etc., which have a high water content and therefore rapidly spoil, would have been consumed first, and their consumption would have had a magical meaning, perhaps related to the function that each serves and therefore relegated to selected members of the M/UP hominin group. This selective consumption has been observed, among others peoples, in Native Americans ones, for which certain parts of the prey were considered appropriate for men or women (Fallon and Enig, 2001). An important fact to consider is the seasonality of the supply of nutrients obtained from hunting resources. For instance, in arctic caribou (Rangifer tarandus granti), severe winter undernourishment has been found, and this leads to the mobilization of large amounts of fat and protein, causing seasonal changes in body mass and composition, so that by late winter up to 45% of fat reserves can be depleted (Gerhart et al., 1996). This would cause a significant energy loss for each prey captured by Palaeolithic hunters, and would force them to make more frequent hunting expeditions to compensate. It should also be borne in mind that the FA composition of adipose tissues in food animals inhabiting extremely cold areas such as reindeer remains relatively stable during winter, while the proportions of principal PUFAs are significantly reduced during spring (Soppela and Nieminen, 2002). This implies that the most critical period for the survival of Palaeolithic hunters must have been spring, when they were not yet able to obtain summer fishing and harvesting resources, and prey provided little fat and with a small percentage of n-3 PUFAs. 3.4. Contribution of different mammals to fulfil the daily need of n3 PUFAs for M/UP hominins In Western Europe, north of the Pyrenees and the Alps, at the M/ UP transition, five animal species dominated, namely: aurochs (Bos primigenius), steppe bison (Bison priscus), horse (Equus caballus), red deer (Cervus elaphus), and reindeer (Rangifer tarandus). Among these, reindeer herds were highly fluctuating, and the availability of this species as prey would have been responsible for the declines in human population densities observed during this period (Morin, 2008). Given that current species are very similar to those existing in the M/UP transition, and their habitats also have close similarity, data on nutritional composition for current free-range species must be similar to that of M/UP species. An important factor to consider when establishing the supply of energy, protein, and n3 PUFA in the various preys is that as meat provides some fat, S/DF contains some amounts of protein, and that such organs are not usually analysed for total FA content. Given the absence of such data, average values have been taken from the USDA Nutrient Database for species related to those hunted at the M/UP transition (Tables S3 and S4). The similarity of values for the various species is remarkable, as indicated by the low SD found. Conversely, meat from all mammals is regularly analysed for fat content (Table S1). The various proportions between fatty tissues (S/DF) and meat to reach the 4500 kcal supply, together with protein, fat, total n-3 PUFA, DHA, EPA þ DHA, and n-3 VLCPUFA values for reindeer are plotted in Fig. 2. Other proteinaceous organs not included in this model, such as

are liver, kidney, spleen, or heart, are comparable to meat, given their similar protein concentration as well as their equivalent FA profiles. However, they constitute only a marginal contribution to total energy intake. Other organs such as intestines and skin have similar protein concentration to meat, but much lower n-3 PUFA amounts. On the other hand, bone marrow can be assimilated to S/ DF, given its high fat content and similar n-3 PUFA profile. Thus, this plot indicates the various amounts of the different n-3 PUFA groups obtained through the daily intake of lean and fatty organs to reach 4500 kcal. The amounts of protein and fat are also represented in the plot. All these quantities have nutritional significance only within the safe protein range, i.e. between the minimum and maximum protein intake possible for good health. Notice that meat and S/DF represent the most abundant proteinaceous and fatty organs, respectively, on which the subsistence of M/UP hominins mainly depended. As displayed in Fig. 2, reaching ~4500 kcal is possible by a daily consumption of between ~20 and ~566 g of meat (safe range) and ~679 to~592 g of S/DF (fatty tissues). This interval is conditioned by protein intake, for which the safe range was between 52 and 162 g daily (Fig. 2), as calculated following Paddon-Jones and Rasmussen (2009) and Bilsborough and Mann (2006) equations, respectively. Both meat and fatty tissues total a maximum daily amount of meal for M/UP hominins of ~1150 g animal food, as mean value for males and females. This value agrees with the observations of Geraci and Smith (1979) for Inuit intake, which reaches ~1200 g daily for males and something less for women, the diet being composed of a mixture of flesh, marrow and selected viscera from several animals. Note that values for total n-3 PUFA, DHA, EPA þ DHA, and n-3 VLCPUFA (EPA þ DPA þ DHA) increases in parallel with meat intake, in which they are abundant. Similar to the plot traced in Fig. 2, respective plots for commonly hunted M/UP mammals have been drawn and the values are summarized in Table S8. In the different M/UP mammals, total n-3 PUFA follows a different trend; it increases in parallel with meat in reindeer and goat, is more or less constant in Bos, and slightly decreases in bison, deer, and horse. However, these latter animals provide a large amount of n-3 PUFA, and therefore this decline is not relevant when choosing to eat meat or S/DF. Even though the daily protein requirement might almost be fulfilled by consuming only S/DF, the maximum meat consumption leads to the maximum VLCPUFA supply, without an appreciable drop in the ALA intake. The choice of the maximum possible consumption of meat by M/UP hunters seems to be well documented, as the processing of the carcass to obtain meat is corroborated at the M/UP transition sites, since the bones of prey usually appear with abundant cut marks, denoting a concerted activity to obtain meat. The assemblages considered here do not contain carnivores or their relative number is insignificant. However, other assemblages contain high percentages of carnivores, such as foxes and wolves. Computing their contributions to M/UP hominin diets is not possible, since there is an absence of data on the FA composition for most of such animals. In any case, using the FA profiles of carnivores to calculate global intakes by Palaeolithic hominins could not be appropriate, given that the capacity of Carnivora to elongate and desaturate lipids is very weak, and thus the FA profiles of their adipose tissues would be very similar to those of the animals they consume. For instance, in experiments with arctic foxes living in reindeer and polar bear areas, it was found that their FA profiles were within the range of values found in the adipose tissue of the latter species (Pond et al., 1995). Thus, the differential contribution of FAs from the carnivorous to the global diet of the hominins could be considered very weak, since they would have been hunted in the same area as the non-carnivorous prey were.

J.L. Guil-Guerrero / Quaternary Science Reviews 157 (2017) 176e187


Fig. 2. The relationship between meat and fatty tissues (S/DF) from reindeer to reach 4500 kcal and thus fulfil the daily energy need for M/UP hominins, considering these organs as the energy source. Fat, protein, and the different n-3 PUFA contained at the various proportions of meat and S/DF are also plotted in the graph. Values for protein and fat for S/DF were 7 g and 70.3 g by 100 g, respectively (Table S3), while values for meat were 1.6 g fat and 22.3 g protein by 100 g, respectively (Table S1). The data for n-3 PUFA of meat and S/DF were taken from Tables S1 and S2, respectively.

3.5. Reconstruction of the overall diet of M/UP hominins using data from several assemblages Although sometimes M/UP populations depend almost exclusively on single food-animal species, in most assemblages skeletal remains from various animals appear, offering the possibility of reconstructing the overall n-3 PUFA intake in each of them. Different nutritional scenarios have been computed for M/UP hominins, as summarized in Table S9. Free-range food animals were selected to determine the carcass weights used to compute the supply of the various n-3 PUFA in the different assemblages. For instance, data for Galician horse (Cabalogalego, 2016), which is considered a relict Palaeolithic species (Guil-Guerrero et al., 2013), while data for red deer were obtained from animals grazing in the field (Wiklund et al., 2001). Computed periods range from latter Mousterian to Magdalenian and thus include both Neanderthals and AMH. Their nutritional status in relation to the various n-3 PUFA intake was assessed taking into account the more recent standards for the various n-3 PUFA intake made by several health organizations. The percentages of compliance for such recommendations at Abri Pataud assemblage over time compared with that of reindeer, auroch, goat and red deer when consumed for optimum n-3 PUFA intake (Fig. 2) are plotted in Fig. 3. Among the values summarized in Table S2, concerning the concentrations of different n-3 PUFAs in different organs of various M/UP mammals, it highlights the contribution made by monogastric animals, such as bear, horse, and mammoth. Other singlestomached mammals such as the woolly rhinoceros, would have made a similar contribution to those considered here, taking into account the similarities in their digestive processes. Thus, the prehistoric assemblages containing bones of monogastric mammals would correspond to scenarios of better nutrition in terms of total n-3 PUFA ingested (Guil-Guerrero et al., 2013, 2014, 2015). However, such contributions concern ALA, and deficiencies of other

n-3 PUFA, (i.e. DHA and EPA) would be roughly similar to those detected in dietary scenarios where ruminant animals dominate. Note that the degree of compliance for the various n-3 PUFA supply varies for the different periods (Table S9), this depending on the relative contribution of each prey. In general, reindeer, goat, and aurochs predominated, and the total n-3 PUFA, EPA, and DHA were deficient (Fig. 3). Moreover, both the DHA and EPA þ DHA supply were also far less than desirable in all the periods considered, regardless of the most abundant prey (Fig. 3). For instance, in the saire assemblage (Table S9; Morin, 2008), the EPA þ DHA Saint-Ce supply for the different periods has been computed from 56 to 76 mg daily. These figures are much lower than others estimated for current daily intakes of EPA þ DHA in most countries for adults, which range from 143 to 950 mg daily (Givens and Gibbs, 2008), although figures for young individuals are sometimes lower. However, such individuals might get additional amounts of EPA þ DHA from ALA because of their higher biosynthetic activity. Concerning DHA, deficiency was also clear for all periods considered, and all assemblages provided figures of between ~20 and 45 mg/day, which are much lower than the recommended safe intake of 125e250 mg/day (Fig. 3). By contrast, in most developed countries, the daily intake of DHA ranges from 184 to 473 mg/day, while in low-income countries the intake is about 96 mg/day (Forsyth et al., 2016). Either way, life is compatible with reduced intake of both EPA and DHA. For instance, Welch et al. (2010), in a study on dietary habit (fish-eaters and non-fish-eating meat-eaters, vegetarians, or vegans) found an average daily intake of DHA of 160 mg, whereas, in non-fish eaters the daily supply of DHA was extreme low, especially in vegans. In relation to this finding, it is necessary to consider that people who adopt vegan lifestyles usually do so after becoming adults, and therefore the effects of the DHA deficit on development remains unknown. Anyway, EPA and DHA are needed for proper foetal development, including neuronal, retinal, and immune function, and their deficiency may affect many


J.L. Guil-Guerrero / Quaternary Science Reviews 157 (2017) 176e187

Fig. 3. Contribution of Abri Pataud faunal assemblage to fulfil the daily need of n-3 PUFAs for M/UP hominins compared with that of reindeer, auroch, goat and red deer. n-3 PUFA intake recommendations for general adult population are: total n-3 PUFA daily intake: 1%E, WHO/FAO (2002) and Nordic Council of Ministers (2013), and 0.5%E, D-A-CH (2008); DHA, 125 mg, half of recommendation for EPA þ DHA (AFFSA, 2010) made by EFSA (2010), and 250 mg, AFFSA (2010); EPA þ DHA, 250 mg, EFSA (2010) and Chinese Nutrition Society (2013), and 500 mg, AFFSA (2010) and Vannice and Rasmussen (2014); n-3 VLCPUFA, 250 mg/day, German Society for Nutrition (2015), and 500 mg, Kris-Etherton and Innis (2007).

aspects of cardiovascular function including inflammation, peripheral artery disease, major coronary events, anticoagulation, poor foetal development, and risk of the development of Alzheimer's disease (Swanson et al., 2012). Concerning DHA, deficiency would have led to a decline in DHA content of the frontal cortex, oxidative damage, impairment of cognitive performance during aging, and age-related declines in neural function, among other adverse health effects. (Yurko-Mauro et al., 2010; Cardoso et al., 2016). The total n-3 VLCPUFA supply seems to have been satisfactory in most periods, due to the high contribution of DPA from mammal meat, although it was deficient in some scenarios, for instance in saire site, from the Proto-Aurignacian to the Evolved Saint-Ce Aurignacian EJJ assemblages (Table S9; Morin, 2008), where n-3 VLCPUFA intake was between 138 and 176 mg/day, which are lower than the recommended safe intake of 250-500 mg/day (Fig. 3). In this regard, in vivo studies have shown limited conversion of DPA to DHA, but its retro-conversion to EPA is evident in a great number of tissues. In any case, DPA could not supply DHA for important brain functions (Kaur et al., 2011). Therefore, the effects of high DPA concentrations in the diet are unclear, and thus it is questionable to include DPA in recommendations for n-3 PUFA intake because DPA is currently a research issue with limited evidence from randomized controlled trial studies (FAO/WHO, 2008). There are important facts to consider to give an overall picture of the computed n-3 PUFA deficiency: i) although different mammals appear in all assemblages, the periods of exclusive consumption of reindeer and other n-3 PUFA-deficient species should have been frequent and over more or less long periods of time, and such consumption would have resulted in very prolonged and severe deficits of all n-3 PUFA types (Fig. 3); ii) in the period considered, most meat could have been consumed roasted, which significantly increases the percentages of SFA and MUFA and decreases the relative proportions of all PUFAs in meat (Alfaia et al., 2010), and thus this could have greatly reduced the availability of n-3 PUFA for hominins; iii) all fatty tissues and meats have a higher percentage of n-6 than n-3 PUFAs, as occurs for example in reindeer meat, in

which the n-6:n-3 ratio is 6:1 (Sampels et al., 2004), and such a ratio hinders the bioavailability of n-3 PUFAs (Guil-Guerrero et al., 2007); iv) the n-3 PUFA scenarios have been computed as the best conditions for n-3 PUFA-rich organs intake dthat is, for meat and fatty tissues in the optimum ratio, and excepting the brain, which is a small organ that would surely have been unevenly distributed among the members of the hunting group. Consumption of other organs, such as bone marrow and intestines, would significantly reduce the daily intake of n-3 PUFAs. Thus, in the scenario described, the n-3 PUFA deficiency could have been much more critical than that computed, and hominins would have had serious difficulties surviving in long intergenerational-deficit periods, especially adults and older individuals. In this sense, nutrient deficiency has been shown to alter the epigenome, which can influence the genetic regulation of key pathways in development and disease, and defects in epigenetic regulation can explain the mechanism of intergenerational transmission of disease caused by poor nutrition (Burdge et al., 2011; Roseboom and Watson, 2012). 3.6. Ascorbic acid deficiency at the M/UP transition Game meat has been suggested to be a good source of vitamin C for M/UP hunters in northern Europe, at least during the long winter periods (Nestle, 1999). However, available data (Table S10) indicate a very different situation, given that both game meat and large viscera of M/UP mammals contain minimal amounts of ascorbic acid; only the brain and kidneys of these animals could be considered to be sources of this vitamin, but their small size prevents the organs from being equitably distributed among the human group, and concentrations are clearly insufficient to avoid scurvy among M/UP hunters. Confirming this, some human bone at the M/UP transition showed symptoms attributable to scurvy, among other causes. For instance, scurvy's cranial symptoms consist of porous and hypertrophic lesions of the vault, affecting frontal and parietal bosses, and related signs have been discovered in Palaeolithic hominins bones (Ortner and Ericksen, 1997;

J.L. Guil-Guerrero / Quaternary Science Reviews 157 (2017) 176e187

Teschler-Nicola et al., 2006; Vercellotti et al., 2010; Eddie, 2013; €mper et al., 2012). Holzka Given the low ascorbic acid values for reindeer organs (Table S10), it can be argued that its prevalence as animal food clearly leads to both an n-3 PUFA deficiency and the onset of scurvy. This fact agrees with Levine (1940), who described the occurrence of subacute scurvy in several Inuit children in Alaska, when consuming frozen reindeer for several days. The intake of raw, recently hunted reindeer might provide a maximum of ~14 mg of ascorbic acid daily (Table S10) considering the supply by both S/DF and meat at the higher intake of protein here established. This amount is at the limit for the appearance of scurvy (Geraci and Smith, 1979), and thus situations such as those of frozen and/or cooked reindeer consumption, which could have been much more than usual among M/UP hunter's hominins, led to chronic subacute scurvy. 3.7. Nutritional role of cannibalism for M/UP hominins The term “cannibalism”, meaning the eating of human bodies or body parts by other humans, has been cited for different reasons, including nutritional, pathological, political, and ritual ones (Lukaschek, 2001). Among well-documented cases of cannibalism in the Palaeolithic, its emergence along the M/UP transition has been widely reported (Table S11). Cannibalism has been linked to spongiform encephalopathy transmission in Neanderthals and their subsequent extinction (Chiarelli, 2004), and it has also been pointed out that this behaviour could have been the adaptive filter for the selection of the heterozygotes for the M129V polymorphism, which confers resistance to kuru (Brookfield, 2003). However, controversy exists about whether the striations on human bone made by Neanderthals were a consequence of defleshing for meat consumption or motivated by secondary burial rather than cannibalism (Russell, 1987). Probably, the intentionality of defleshing will always be interpreted from different points of view; however, cannibalism has never been interrupted from the Palaeolithic until current humans, through different evolutionary periods (Table S11), and evidence for cannibalism has been found in human coprolites, revealing human myoglobin remains as unequivocal proof of human meat intake (Marlar et al., 2000). When the target of the cannibalistic act was the meat itself, this practice invariably made visible marks on the bones; but when the aim was the consumption of other organs such as liver or S/DF, bones will not show the typical cut-marks caused by defleshing. Therefore, such a practice could have been much more common than evidence shows. Meat was possibly more or less abundant for M/UP hunters, given that most prey yield more meat than fatty tissues, and the latter was needed to a much greater extent than meat for subsistence, and therefore the main target for cannibals must have been fatty organs such as S/DF, instead of meat, which would have occurred without leaving marks on the bones. Maybe the dependence on a single species such as reindeer for subsistence has always been a situation to instinctively avoid by M/UP hunters. At such stages, cannibalism possibly would have increased. The n-3 PUFA content in the main human organs is summarized in Table S5. Note the high amounts of all n-3 classes that all human organs contain at higher figures than the corresponding M/UP mammals. Fig. 4 shows a comparison of the DHA content between the main organs of humans and food animals, using data from Tables S1, S2, and S5. The differences in concentrations are striking. In any case, it is possible that, in situations of n-3 PUFA deficiency, some human organs could diminish their PUFA concentrations, as probably would occur with depot fats, although this possibility is unconfirmed, and data for human S/DF (Table S5) are quite conclusive.


Unlike what happens to food animals, the human brain reaches a large size (~1.5 kg), and given its high DHA concentration (1.6 g/ 100 g), in a single brain the total DHA accounted for ~24 g, providing ~2450 kcal. With such low caloric content, it cannot be considered an energy organ. It would have been immediately consumed like all other human viscera, given its high perishability, although, when cooked, it could have lasted several days to be consumed as a kind of nutritional supplement. Considering similarities between current cannibals and the earliest ones, such as current South Fore cannibals, the human brain could always have been considered a delicacy, with restrictive rules regarding consumption. For instance, South Fore females of all ages consumed it, whereas males over the age 6 never did (Whitfield et al., 2008). Thus, in the Palaeolithic, it is likely that this organ could have played a role as DHA-rich supplement to be consumed by individuals with special needs, such as pregnant women or aged individuals. Considering human S/DF, it could also have provided suitable amounts of DHA to M/UP hunters, given that the consumption of about 50 g of human S/DF daily is enough to fulfil the daily need of DHA for any member of the M/UP group (Table S5). Logically, the supply of n-3 PUFA enriched human organs would have been more or less occasional, but the human bodies (those obtained from natural death and some others from ritual sacrifices or captures) could have been preserved under the snow in the long winter of the Ice Ages, to be eaten on a regular basis. The bodies of dead children would also be in such deposits, and, given that the M/ UP transition was a time of very high birth rates, in this case a fertility rate of about 50 per thousand (Sattenspiel and Harpending, 1983), such supply could have been quite regular. This means that cannibalism practice would have been similar to that which occurred during the 1972 Andes flight disaster, in which the bodies of passengers that died in the crash were preserved by survivors under the snow to be eaten on a daily basis. By contrast, during the short summers of the Ice Age, in thawing periods, instead of using such rich n-3 PUFA source, M/UP hominins might have consumed some plants containing more or less ascorbic acid and some n-3 C18-PUFA amounts, as well as some freshwater fish, which contain enough n-3 VLCPUFA amounts. This behaviour has been described in Inuit populations, which use some plant foods in the summer to supplement their diets, thus obtaining considerable amounts of ascorbic acid in this season (Geraci and Smith, 1979). On the other hand, unlike Palaeolithic hominins, Inuit people has regularly available all year seals and fish for hunting and fishing (Geraci and Smith, 1979), which are notable n-3 VLCPUFA sources, so Inuit people would never had suffered deficiency symptoms of such nutrients. The ascorbic acid content of several human organs is summarized in Table S10, while a comparison of ascorbic acid concentrations between food-animals and human organs is given in Fig. 5. As with the case of n-3 PUFA, all human organs are more or similarly rich in ascorbic acid compared with those of food-animals, thus able to supplement diets, although in scurvy situations such concentrations might decline to some extent. As seen, the human body contains valuable nutrients, and without cannibalism practice M/UP hominins probably might not have overcome some long-term high-deficiency-periods, especially human groups depending on reindeer, as explained above. Such a state of deficiency is well characterized, and changes made in the computed model, such as percentages of S/DF or meat consumed, caloric needs or weight estimated for M/UP hominins, result in small variations in the percentages of n-3 PUFAs ingested daily. The evolutionary advantages of cannibalism could have been very subtle. For instance, today evidence indicates that Neanderthals had capacities similar to those of modern humans, and their potential to establish complex social structures have been reported


J.L. Guil-Guerrero / Quaternary Science Reviews 157 (2017) 176e187

Fig. 4. DHA content in organs of food-animals (mean values for all species) compared with human ones. Values are taken from Tables S1, S2, and S5.

Fig. 5. Ascorbic acid content of some human and food-animal organs. Values are taken from Table S10.

(Fahlander, 2013). The older members of the hominin groups, both Neanderthals and AMH, were needed in M/UP cultures, as some individuals retain cultural knowledge and transmit it, such as hunting areas or technology, to the rest of the social group. Their importance is clear, because such individuals received special treatment in death (Fahlander, 2013). This suggests that at least some of the older individuals were recognized to have some special position, indicating social privilege for both sexes, as the aged people could have been decisive for the survival of M/UP hominins. Some of the functions of such individuals could have been: - Preserving group identity and unity by transmitting mythology

and relationships with neighbouring human groups. - Arbitrating the application of social norms, taboos, and laws. - Showing the adequate use of some foods, medicinal plants, etc. - Explaining supernatural forces, such as lightning, earthquakes, eclipses, etc. to eliminate the anxiety of the group. - Remembering the location of seasonal hunting grounds. - Maintaining stone technologies, among others. However, aged people could have had critical difficulties surviving due to their n-3 PUFA metabolism, since, unlike younger

J.L. Guil-Guerrero / Quaternary Science Reviews 157 (2017) 176e187

individuals, older individuals do not properly metabolise ALA to DHA, due to the previously exposed D-desaturase impairment (Harris, 2012; Bradbury, 2011). The total n-3 PUFAs proved more or less sufficient in some periods for the survival of young individuals, considering data for some assemblages in which the hunted animals varies. Furthermore, consumption of DHA-enriched fatty human organs by elderly M/UP hominins could have been vital in order to maintain a good level of mental and physical activity for aged individuals, thus leading to the survival of the whole group. On the other hand, cannibalistic practice would have had a strong seasonal component. As discussed above, the most critical period for the survival of M/UP hunters from the standpoint of the nutrient deficits would be in the spring, when the n-3 PUFA content of S/DF of their prey were depleted, and n-3 PUFA summer resources from fishing and gathering were not yet available. In this season, in addition to the bodies obtained from natural deaths, some members of the hominin group would probably have been selected for ritual sacrifices to be eaten. Perhaps this situation could be related to the origin of symbolism associated with totemic meals and related moral and religious implications. Finally, it bears considering that cannibalism would be a preadaptive behaviour, given that it has also been reported in primates, as in wild chimpanzees (Nishida and Kawanaka, 1985). Therefore, cannibalism would have had a selective factor influencing the evolution of human culture for hominins at the M/UP transition, which is the period considered here due to the observed increase of the cannibalism in that period, which took place in parallel with the greater abundance of the reindeer in the Palaeolithic assemblages in Europe. However, less markedly, this behaviour also took place in the Middle Palaeolithic. Interpreting the essence of cannibalism an act of violence in humans but with social and institutional roots, we could infer that this stems from an innate desire to gain critical nutrients, which results in the death and nutritional assimilation of other individuals, whether group members or outsiders. Ritual violence, evidenced by cannibalism, and spiritual expressions manifested through burial have been a typical human expression in the Gravettian, more than in the Aurignacian, and thus it is thought that such behaviour evolved throughout the Upper Palaeolithic (Pearce et al., 2014). Our society has many allegories and symbols that recreate our cannibal past: religious, artistic, political, and others, which can be easily detected. An example is religious symbolism involving the assimilation of any divinity through food or drink intake.

4. Conclusions A situation of chronic deficiency of some essential nutrients for hominins at the M/UP transition in Europe is described in this work. These nutrients include some n-3 PUFAs and ascorbic acid, which occurred to a limited degree in regularly consumed foods by hominins in that period. By appropriate calculations applied both to single food-animals and to their assemblages appearing in several human M/UP sites, it was found that when strong dependence periods for some food-animals, as for reindeer, n-3 PUFA and ascorbic acid intake was clearly deficient. To solve this, M/UP hominins might have developed a cannibalistic behaviour. This is so because all organs of the human body are rich sources of n-3 PUFA and ascorbic acid, which could have helped to improve the nutritional status of hominins during such times, thus enabling their survival. A consequence of this situation might have been the institutionalization of social violence and symbolic rituals involving human sacrifices, especially in the spring, in order to obtain human tissues for consumption.


Acknowledgments The author wish to acknowledge the excellent critical review of the manuscript made by Dr. Federico García-Maroto and Dr. Diego pez-Alonso, from the University of Almería. Lo Appendix A. Supplementary data Supplementary data related to this article can be found at http:// dx.doi.org/10.1016/j.quascirev.2016.12.016. References curite  sanitaire des aliments relatif a  AFFSA, 2010. AVIS de l’Agence française de se s pour les acides gras (accesed l’actualisation des apports nutritionnels conseille 15.07.16). www.anses.fr/fr/system/files/NUT2006sa0359.pdf. Alfaia, C.M., Alves, S.P., Lopes, A.F., Fernandes, M.J., Costa, A.S., Fontes, C.M., , et al.Prates, J.A., 2010. Effect of cooking methods on fatty acids, conjugated isomers of linoleic acid and nutritional quality of beef intramuscular fat. Meat Sci. 84 (4), 769e777. Bilsborough, S., Mann, N., 2006. A review of issues of dietary protein intake in humans. Int. J. Sport. Nutr. Exerc. Metab. 16, 129. Binford, L.R., 1984. Faunal Remains from Klasies River Mouth. Academic Press, NewYork. Bocherens, H., Drucker, D.G., Billiou, D., Patou-Mathis, M., Vandermeersch, B., 2005. saire I NeanIsotopic evidence for diet and subsistence pattern of the Saint-Ce derthal: review and use of a multi-source mixing model. J. Hum. Evol. 49, 71e87. Bocherens, H., Drucker, D.G., Madelaine, S., 2014. Evidence for a 15 N positive excursion in terrestrial foodwebs at the Middle to Upper Palaeolithic transition in south-western France: implications for early modern human palaeodiet and palaeoenvironment. J. Hum. Evol. 69, 31e43. Bradbury, J., 2011. Docosahexaenoic acid (DHA): an ancient nutrient for the modern human brain. Nutrients 3, 529e554. Brookfield, J.F., 2003. Human evolution: a legacy of cannibalism in our genes? Curr. Biol. 13, 592e593. Burdge, G.C., Calder, P.C., 2005. a-Linolenic acid metabolism in adult humans: the effects of gender and age on conversion to longer-chain polyunsaturated fatty acids. Eur. J. Lipid Sci. Technol. 107, 426e439. Burdge, G.C., Finnegan, Y.E., Minihane, A.M., Williams, C.M., Wootton, S.A., 2003. Effect of altered dietary n-3 fatty aid intake upon plasma lipid fatty acid composition, conversion of [13C]a-linolenic acid to longer-chain fatty acids and partitioning towards b-oxidation in older men. Br. J. Nutr. 90, 311e321. Burdge, G.C., Hoile, S.P., Uller, T., Thomas, N.A., Gluckman, P.D., Hanson, M.A., Lillycrop, K.A., 2011. Progressive, transgenerational changes in offspring phenotype and epigenotype following nutritional transition. In: Imhof, A. (Ed.), PLoS One 6, e28282. Calder, P.C., 2012. Mechanisms of action of (n-3) fatty acids. J. Nutr. 142, 592e599. Calder, P.C., 2015. Functional roles of fatty acids and their effects on human health. J. Parenter. Enter. Nutr. 39, 18e32. Calviello, G., Su, H.M., Weylandt, K.H., Fasano, E., Serini, S., Cittadini, A., 2013. Experimental evidence of-3 polyunsaturated fatty acid modulation of inflammatory cytokines and bioactive lipid mediators: their potential role in inflammatory, neurodegenerative, and neoplastic diseases. Biomed. Res. Int. 2013, 743171. Cardoso, C., Afonso, C., Bandarra, N.M., 2016. Dietary DHA and health: cognitive function ageing. Nutr. Res. Rev. 1e14. Chiarelli, B., 2004. Spongiform encephalopathy, cannibalism and Neanderthals extinction. J. Hum. Evol. 19, 81e91. Chinese Nutrition Society, 2013. Chinese DRIs Handbook. Cordain, L., Eaton, S.B., Brand Miller, J., Mann, N., Hill, K., 2002. Original communications-the paradoxical nature of hunter-gatherer diets: meat-based, yet non-atherogenic. Eur. J. Clin. Nutr. 56, 42. s-Sa nchez, M., Morales-Mun ~ iz, A., Simo  n-Vallejo, M.D., Lozano-Francisco, M.C., Corte nezVeraPel aez, J.L., Finlayson, C., Rodríguez-Vidal, J., Delgado-Huertas, A., Jime Espejo, F.J., Martínez-Ruiz, F., Aranzazu Martínez-Aguirre, M., PascualBergad Granged, A.J., Merce a-Zapata, M., Gibaja-Bao, J.F., Riquelme-Cantal, J.A.,  pez-Sa ez, J.A., RodrigoG Lo amiz, M., Sakai, S., Sugisaki, S., Finlayson, G., Fa, D.A., Bicho, N.F., 2011. Earliest known use of marine resources by Neanderthals. PloS One 6, e24026. Craig, O.E., Biazzo, M., Colonese, A.C., Di Giuseppe, Z., Martinez-Labarga, C., Vetro, D.L., Lelli, R., Martini, F., Rickards, O., 2010. Stable isotope analysis of Late Upper Palaeolithic human and faunal remains from Grotta del Romito (Cosenza), Italy. J. Arch. Sci. 37, 2504e2512. Crawford, M.A., Sinclair, A.J., 1976. The long chain metabolites of linoleic and linolenic acids in liver and brains of herbivores and carnivores. Comp. Biochem. Physiol. B 54, 395e401. Crawford, M.A., Bloom, M., Broadhurst, C.L., Schmidt, W.F., Cunnane, S.C., Gallim, C., Gehbremeskel, K., Linseisen, F., Lloyd-Smith, J., Parkington, J., 1999. Evidence for the unique function of docosahexaenoic acid during the evolution of the modern hominid brain. Lipids 34, 39e47.


J.L. Guil-Guerrero / Quaternary Science Reviews 157 (2017) 176e187

Crile, G., Quiring, D.P., 1940. A record of the body weight and certain organ and gland weights of 3690 animals. Ohio J. Sci. 40, 219e259. € €hrung, Osterreichische D-A-CH, 2008. Deutsche Gesellschaft für Erna Gesellschaft €hrung, Schweizerische Gesellschaft für Erna €hrungsforschung, Schweifür Erna €hrung: Referenzwerte für die Na €hrstoffzufuhr. zerische Vereinigung für Erna Umschau/Braus Verlag, Frankfurt. Dolan, L.C., Matulka, R.A., Burdock, G.A., 2010. Naturally occurring food toxins. Toxins 2, 2289e2332. Drucker, D.G., Henry-Gambier, D., 2005. Determination of the dietary habits of a Magdalenian woman from Saint-Germain-la-Riviere in southwestern France using stable isotopes. J. Hum. Evol. 49, 19e35. Eddie, D.M., 2013. Examination of Trauma in a Neandertal Ulna. Doctoral dissertation. University of Kansas. EFSA Panel on Dietetic Products, Nutrition, and Allergies (NDA), 2010. Scientific opinion on dietary reference values for fats, including saturated fatty acids, polyunsaturated fatty acids, monounsaturated fatty acids, trans fatty acids, and cholesterol. EFSA J. 8, 1461. Fahlander, F., 2013. Intersecting generations: burying the old in a Neolithic hunterfisher community. Camb. Archaeol. J. 23, 227e239. Fallon, S., Enig, M.G., 2001. Guts and grease: the diet of native Americans. Wise Tradit. 40e47. FAO/WHO, 2008. Expert Consultation on Fats and Fatty Acids in Human Nutrition, November 10e14, 2008. WHO HQ, Geneva. Forsyth, S., Gautier, S., Salem Jr., N., 2016. Global estimates of dietary intake of docosahexaenoic acid and arachidonic acid in developing and developed countries. Ann. Nutr. Metab. 68, 258e267. , E., Martínez-Moreno, J., Mora, R., Nún ~ ez, M., Richards, M.P., 2009. Garcia-Guixe Stable isotope analysis of human and animal remains from the Late Upper Palaeolithic site of Balma Guilany a, southeastern Pre-Pyrenees, Spain. J. Archaeol. Sci. 36, 1018e1026. Georgiadi, A., Kersten, S., 2012. Mechanisms of gene regulation by fatty acids. Adv. Nutr. Int. Rev. J. 3, 127e134. Geraci, J.R., Smith, T.G., 1979. Vitamin C in the diet of Inuit hunters from holman, Northwest territories. Arctic 135e139. Gerhart, K.L., White, R.G., Cameron, R.D., Russell, D.E., 1996. Body composition and nutrient reserves of arctic caribou. Can. J. Zool. 74, 136e146. €vention ausgewa €hlter German Society for Nutrition (DGE), 2015. Fettzufuhr und Pra ern€ ahrungsmitbedingter Krankheiten. http://www.dge.de/fileadmin/public/ doc/ws/ll-fett/v2/Gesamt-DGE-Leitlinie-Fett-2015.pdf (Accessed 12 September 2016). Givens, D.I., Gibbs, R.A., 2008. Current intakes of EPA and DHA in European populations and the potential of animal-derived foods to increase them. Proceed. Nutr. Soc. 67, 273e280. Gray, R.G., Kousta, E., McCarthy, M.I., Godsland, I.F., Venkatesan, S., Anyaoku, V., Johnston, D.G., 2013. Ethnic variation in the activity of lipid desaturases and their relationships with cardiovascular risk factors in control women and an atrisk group with previous gestational diabetes mellitus: a cross-sectional study. Lipids Health Dis. 12, 25. Grayson, D.K., Delpech, F., 2002. Specialized early Upper Palaeolithic hunters in southwestern France? J. Archaeol. Sci. 29, 1439e1449. Guil-Guerrero, J.L., 2007. Stearidonic acid (18:4n-3): metabolism, nutritional importance, medical uses and natural sources. Eur. J. Lipid Sci. Tech. 109, 1226e1236. n-Cervera, M.A., Venegas-Venegas, C.E., Ramos-Bueno, R.P., Guil-Guerrero, J.L., Rinco rez-Medina, M.D., 2013. Highly bioavailable a-linolenic acid from the subSua cutaneous fat of the Palaeolithic Relict “Galician horse”. Int. Food Res. J. 20, 3249e3258. Guil-Guerrero, J.L., 2014. Common mistakes about fatty acids identification by gaseliquid chromatography. J. Food Compos. Anal. 33, 153e154. Guil-Guerrero, J.L., Tikhonov, A., Rodríguez-García, I., Protopopov, A., Grigoriev, S., Ramos-Bueno, R.P., 2014. The fat from frozen mammals reveals sources of essential fatty acids suitable for Palaeolithic and Neolithic humans. PloS One 9 (1), e84480. Guil-Guerrero, J.L., Rodríguez-García, I., Kirillova, I., Shidlovskiy, F., RamosBueno, R.P., Savvinov, G., Tikhonov, A., 2015. The PUFA-enriched fatty acid profiles of some frozen Bison from the early holocene found in the siberian permafrost. Sci. Rep. 5. http://cabalogalego.com/(Accessed 22 October 2016) Hardy, B.L., Moncel, M.H., 2011. Neanderthal use of fish, mammals, birds, starchy plants and wood 125e250,000 years ago. PloS One 6, e23768. Harris, W.S., 2012. Stearidonic acid as a ‘pro-eicosapentaenoic acid’. Curr. Opin. Lipidol. 23, 30e34. Hawkes, K., O'Connell, J.F., 1985. Optimal foraging models and the case of the. Kung. Am. Anthropol. 87, 401e405. Hermanussen, M., 2003. Stature of early europeans. Horm. (Athens) 2, 175e178. Hockett, B., Haws, J., 2003. Nutritional ecology and diachronic trends in Paleolithic diet and health. Evol. Anthropol. 12, 211e216. €mper, J., Kretschmer, I., Maier, A., 2014. The upper-late palaeolithic transition Holzka in western central Europe. Typology, technology, environment and demog€srath, 21ste24th June 2012, vol. raphy. In: Report on the Workshop Held in Ro 36, pp. 161e186. Arch. Inf. Kaplan, H., Hill, K., 1992. Human subsistence behavior. In: Smith, E.A., Winterhalder, B. (Eds.), Evolution, Ecology and Human Behavior, pp. 167e202. Chicago, IL: Aldine. Kaur, G., Cameron-Smith, D., Garg, M., Sinclair, A.J., 2011. Docosapentaenoic acid

(22: 5n-3): a review of its biological effects. Prog. Lipid Res. 50, 28e34. Kris-Etherton, P.M., Innis, S., 2007. American dietetic association and dietitians of Canada position of the american dietetic association and dietitians of Canada: dietary fatty acids. J. Am. Diet. Assoc. 107, 1599e1611. Levine, V.E., 1940. The ascorbic acid content of the blood of the Eskimo. J. Biol. Chem. 133, 399e407. Lukaschek, K., 2001. The History of Cannibalism (Doctoral Dissertation, Thesis Submitted in Fulfillment of the MPhil Degree in Biological Anthropology. University of Cambridge, UK Lucy Cavendish College, Electronic. Marlar, R.A., Leonard, B.L., Billman, B.R., Lambert, P.M., Marlar, J.E., 2000. Biochemical evidence of cannibalism at a prehistoric Pueblo an site in southwestern Colorado. Nature 407, 74e78. Mathers, K., Henneberg, M., 1995. Were we ever that big? Gradual increase in hominid body size over time. Homo 46, 141e173. Miles, E.A., Calder, P.C., 2012. Influence of marine n-3 polyunsaturated fatty acids on immune function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis. Br. J. Nutr. 107, 171e184. Morin, E., 2008. Evidence for declines in human population densities during the early Upper Paleolithic in western Europe. Proc. Natl. Acad. Sci. U.S.A 105, 48e53. Nestle, M., 1999. Animal v. plant foods in human diets and health: is the historical record unequivocal? Proc. Nutr. Soc. 58, 211e218. Nishida, T., Kawanaka, K., 1985. Within-group cannibalism by adult male chimpanzees. Primates 26, 274e284. Nordic Council of Ministers, 2013. Nordic Nutrition Recommendations 2012. Part 1. Nordic Council of Ministers, Copenhagen. Summary, principles and use. Nord 2013; 009. Olmedo, J.M., Yiannias, J.A., Windgassen, E.B., Gornet, M.K., 2006. Scurvy: a disease almost forgotten. Int. J. Dermatol. 45, 909e913. Ortner, D.J., Ericksen, M.F., 1997. Bone changes in the human skull probably resulting from scurvy in infancy and childhood. Int. J. Osteoarchaeol 7, 212e220. Paddon-Jones, D., Rasmussen, B.B., 2009. Dietary protein recommendations and the prevention of sarcopenia: protein, amino acid metabolism and therapy. Curr. Opin. Clin. Nutr. Metab. Care 12, 86. Padh, H., 1990. Cellular functions of ascorbic acid. Biochem. Cell Biol. 68 (10), 1166e1173. Pearce, E., Shuttleworth, A., Grove, M., Layton, R., 2014. In: Dunbar, R., Gamble, C., Gowlett, J. (Eds.), The Lucy Project: Benchmark Papers. Oxford University Press, Oxford, pp. 356e379. Pond, C.M., Mattacks, C.A., Gilmour, I., Johnston, M.A., Pillinger, C.T., Prestrud, P., 1995. Chemical and carbon isotopic composition of fatty acids in adipose tissue as indicators of dietary history in wild arctic foxes (A lopex lagopus) on Svalbard. J. Zool. 236 (4), 611e623. Pryor, A.J.E., 2008. Following the fat: food and mobility in the European Upper Palaeolithic 45,000-18,000 ya. In: Lightfoot, E. (Ed.), Movement, Mobility and Migration. Archaeological Review from Cambridge. Academia.edu, Cambridge, pp. 161e180. Reimers, E., 1984. Body composition and population regulation of Svalbard reindeer. Rangifer 4, 16e21. Revedin, A., Longo, L., Lippi, M.M., Marconi, E., Ronchitelli, A., Svoboda, J., Anichini, E., Gennai, E., Aranguren, B., 2015. New technologies for plant food processing in the Gravettian. Quat. Int. 359, 77e88. Richards, M.P., Trinkaus, E., 2009. Isotopic evidence for the diets of European Neanderthals and early modern humans. Proc. Nat. Acad. Sci. 106, 16034e16039. Richards, M.P., Hedges, R.E.M., Jacobi, R., Current, A., Stringer, C., 2000. FOCUS: Gough's Cave and Sun Hole Cave human stable isotope values indicate a high animal protein diet in the British Upper Palaeolithic. J. Archaeol. Sci. 27, 1e3. Richards, M.P., Pettitt, P.B., Trinkaus, E., Smith, F.H., Paunovi c, M., Karavani c, I., 2000b. Neanderthal diet at Vindija and Neanderthal predation: the evidence from stable isotopes. Proc. Nat. Acad. Sci. 97, 7663e7666. Richards, M.P., Taylor, G., Steele, T., McPherron, S.P., Soressi, M., Jaubert, J., Orschiedt, J., Mallye, J.B., Rendu, W., Hublin, J.J., 2008. Isotopic dietary analysis of a Neanderthal and associated fauna from the site of Jonzac (Charente-Maritime), France. J. Hum. Evol. 55, 179e185. Roebroeks, W., 2008. Time for the Middle to upper paleolithic transition in Europe. J. Hum. Evol. 55, 918e926. Roseboom, T.J., Watson, E.D., 2012. The next generation of disease risk: are the effects of prenatal nutrition transmitted across generations? Evidence from animal and human studies. Placenta 33, e40ee44. Rudman, D., DiFulco, T.J., Galambos, J.T., Smith 3rd, R.B., Salam, A.A., Warren, W.D., 1973. Maximal rates of excretion and synthesis of urea in normal and cirrhotic subjects. J. Clin. Inv. 52, 241-2249. Russ, H., 2013. A Taphonomic Approach to Reconstructing Upper Palaeolithic Hunter¿ Gatherer Fishing Strategies. A Load of Old Trout! Doctoral dissertation, University of Bradford. Russell, M.D., 1987. Mortuary practices at the krapina Neandertal site. Am. J. Phys. Anthropol. 72, 381e397. Sampels, S., Pickova, J., Wiklund, E., 2004. Fatty acids, antioxidants and oxidation stability of processed reindeer meat. Meat Sci. 67 (3), 523e532. Sattenspiel, L., Harpending, H., 1983. Stable populations and skeletal age. Am. Antiq. 48, 489e498. Silberbauer, G., 1981. Hunter¼gatherers of the central Kalahari. In: Harding, R.S.O., Teleki, G. (Eds.), Omnivorous Primates. Columbia University Press, New York, pp. 455e498. Simms, S.R., 1987. Behavioural Ecology and Hunter-gatherer Foraging. An example

J.L. Guil-Guerrero / Quaternary Science Reviews 157 (2017) 176e187 from the Great Basin. British Archaeological Reports. International Series, Oxford. Simopoulos, A.P., 2006. Evolutionary aspects of diet, the omega-6/omega-3 ratio and genetic variation: nutritional implications for chronic diseases. Biomed. Pharmacother. 60, 502e507. Snodgrass, J.J., Leonard, W.R., 2009. Neanderthal energetics revisited: insights into population dynamics and life history evolution. PaleoAnthropol 2009, 220e237. Soppela, P., Nieminen, M., 2002. Effect of moderate wintertime undernutrition on fatty acid composition of adipose tissues of reindeer (Rangifer tarandus tarandus L.). Comp. Biochem. Physiol. Part A Mol. Integr. Physiol. 132 (2), 403e409. Sorensen, M.V., Leonard, W.R., 2001. Neandertal energetics and foraging efficiency. J. Hum. Evol. 40, 483e495. Speth, J.D., Spielmann, K.A., 1983. Energy source, protein metabolism, and huntergatherer subsistence strategies. J. Anthropol. Archaeol. 2, 1e31. Steegmann, A.T., Cerny, F.J., Holliday, T.W., 2002. Neanderthal cold adaptation: physiological and energetic factors. Am. J. Hum. Biol. 14, 566e583. Stiner, M.C., 1991. Food procurement and transport by human and non-human predators. J. Archaeol. Sci. 18, 455e482. Swanson, D., Block, R., Mousa, S.A., 2012. Omega-3 fatty acids EPA and DHA: health benefits throughout life. Adv. Nutr. 3, 1e7. Teschler-Nicola, M., Czerny, C., Oliva, M., Schamall, D., Schultz, M., 2006. Pathological alterations and traumas in the human skeletal remains from Mlade c. In: Early Modern Humans at the Moravian Gate. Springer, Vienna, pp. 473e489. Vannice, G., Rasmussen, H., 2014. Position of the academy of nutrition and dietetics: dietary Fatty acids for healthy adults. J. Acad. Nutr. Diet 114, 136e153. Vercellotti, G., Caramella, D., Formicola, V., Fornaciari, G., Larsen, C.S., 2010. Porotic


hyperostosis in a late upper palaeolithic skeleton (Villabruna 1, Italy). Int. J. Osteoarchaeol. 20, 358e368. Wainwright, P.E., 2002. Dietary essential fatty acids and brain function: a developmental perspective on mechanisms. Proc. Nutr. Soc. 61, 61e69. Welch, A.A., Shakya-Shrestha, S., Lentjes, M.A., Wareham, N.J., Khaw, K.T., 2010. Dietary intake and status of n-3 polyunsaturated fatty acids in a population of fish-eating and non-fish-eating meat-eaters, vegetarians, and vegans and the precursor-product ratio of a-linolenic acid to long-chain n-3 polyunsaturated fatty acids: results from the EPIC-Norfolk cohort. Am. J. Clin. Nutr. 92, 1040e1051. Whitfield, J.T., Pako, W.H., Collinge, J., Alpers, M.P., 2008. Mortuary rites of the south Fore and kuru. Philos. Trans. R. Soc. Lond. B Biol. Sci. 363, 3721e3724. WHO/FAO Expert Consultation on Diet, Nutrition and the Prevention of Chronic Diseases, 2002. Diet, Nutrition and the Prevention of Chronic Diseases: Report of a Joint WHO/FAO Expert Consultation, Geneva, 28 January -1 February 2002. WHO technical report series 916. Wiklund, E., Stevenson-Barry, J.M., Duncan, S.J., Littlejohn, R.P., 2001. Electrical stimulation of red deer (Cervus elaphus) carcassesdeffects on rate of pHdecline, meat tenderness, colour stability and water-holding capacity. Meat Sci. 59, 211e220. , M., Naito, Y.I., Semal, P., Wißing, C., Rougier, H., Crevecoeur, I., Germonpre Bocherens, H., 2015. Isotopic evidence for dietary ecology of late Neandertals in North-Western Europe. Quat. Int. 411, 327e345. Yurko-Mauro, K., McCarthy, D., Rom, D., Nelson, E.B., Ryan, A.S., Blackwell, A., Salem, N., Stedman, M., 2010. Beneficial effects of docosahexaenoic acid on cognition in age-related cognitive decline. Alzheimer’s Dement. 6, 456e464.