Compound-specific stable carbon isotopic detection of pig product processing in British Late Neolithic pottery.

Compound-specific stable carbon isotopic detection of pig product processing in British Late Neolithic pottery. By extracting lipids from potsherds and determining the[[delta][sup.13.C]] of the most abundant fatty acids, degraded fats fromruminant ruminant,any of a group of hooved mammals that chew their cud, i.e., that regurgitate and chew again food that has already been swallowed. Ruminants have an even number of toes on each foot and a stomach with either three or four chambers. animals, such as cattle, and non-ruminant animals, such aspigs, can be distinguished. The authors use this phenomenon toinvestigate Late Neolithic pig exploitation and find that the pig'signature' was more frequently found among residues fromGrooved Ware than other prehistoric pottery types. Keywords: Neolithic, Britain, archaeological chemistry, fattyacids, Groove d Ware, pigs, residues, ruminants, TAGs Introduction It is now well established that unglazed pottery vessels possessthe capacity to absorb substantial concentrations of fats, oils andwaxes (i.e. lipids) during the processing of animal and plant products(Beck et al. 1989; Evershed et al. 1991; 1992; 1994; 2002; Charters etal. 1993; 1995; 1997; Dudd & Evershed 1998; Mottram et al. 1999;Evershed 2000; Copley et al. 2001a; 2001b; 2003; 2005a; 2005b; 2005c;2005d; 2005e; Hansel han¡¤sel?n. & v.Variant of handsel. et al. 2008; Reber et al. 2004). Significantly,degraded animal fats are the most commonly observed components, raisinga number of possibilities for using absorbed residues in pottery as analternative source of information relating to the exploitation of bothdomestic and wild species (Evershed et al. 1999; Copley et al. 2003;2005d; Mukherjee et al. 2005). However, while degraded animal fats arereadily detected, the animal species from which they derive is moredi fficult to determine. The chromatograms displayed in Figure 1 show howdegraded ruminant adipose adipose/ad¡¤i¡¤pose/ (ad¡äi-pos)1. fatty.2. the fat present in the cells of adipose tissue.ad¡¤i¡¤poseadj.Of, relating to, or composed of animal fat; fatty. , dairy and porcine porcine/por¡¤cine/ (por¡äsin) pertaining to swine. porcinepertaining to pig. See also hog (1), swine.porcine circovirus 1a nonpathogenic virus. adipose fats resemble eachother chemically. Triacylglycerols (TAGs) are the major constituents of flesh animalfats and their distributions can be diagnostic to species level. Studiesof modern animal fats have shown that bovine adipose fats containsaturated TAGs of total acyl ac¡¤yln.A organic radical having the general formula RCO, derived from the removal of a hydroxyl group from an organic acid.acyl1. an organic radical derived from a fatty acid by removal of the hydroxyl group.2. carbon numbers between [C.sub.42] and[C.sub.54], similarly, ovine ovinepertaining t o, characteristic of, or derived from sheep.ovine atopic dermatitissymmetrical erythema, alopecia, lichenification, excoriation on woolless areas; sporadic cases, recur each summer. adipose fats contain TAGs ranging from[C.sub.44] (trace) to [C.sub.54]. Artificially degraded dairy fats havea characteristically wide TAG distribution ([C.sub.40]-[C.sub.54]),whereas for porcine fats it is 'narrow' ([C.sub.44]-[C.sub.54]but with very low abundances of [C.sub.44], [C.sub.46] and [C.sub.54];Dudd 1999). Recognisable distributions have been found to persist inanimal fats extracted from archaeological potsherds (Figure 2; Dudd& Evershed 1998; Kimpe et al. 2001; 2002), although there aredifferences in the abundances of individual TAGs between modern andancient animal fats (Figure 2). This is probably due to alteration ofthe more chemically labile labile/la¡¤bile/ (la¡äbil)1. gliding; moving from point to point over the surface; unstable; fluctuating.2. chemically unstable.la¡¤bile adj.1. and water soluble/volatile constituentstaking place during cooking and degradation (Dudd 1999). Burialconditions at the vast majority of sites are not conducive to TAGpreservation and over archaeological timescales they become hydrolysedto form di- and monoacylglycerols (DAGs and MAGs) and free fatty acids,with the latter usually becoming the major constituents of lipidresidues. Hence, alternative means of animal fat identification have tobe sought. [FIGURE 1 OMITTED] A more reliable method for distinguishing degraded animal fats iscompound-specific stable carbon isotope analysis to determine stablecarbon isotope ratios [[delta][sup.13.C]] values; Evershed et al. 1994;1997; 2002; 2003; Mottram et al. 1999) of individual fatty acids. Wehave shown that while the original TAGs are hydrolysed over time the[[delta][sup.13.C] values of their constituent free fatty acids are notaffected significantly by degradation (Evershed et al. 1999). Thescatter plot presented in Figur e 3 shows [[delta][sup.13.C]] values of[C.sub.16:0] and [C.sub.18:0] fatty acids of modern reference animalfats (bovine, ovine and porcine adipose and bovine milk) from animalsraised on isotopically comparable diets to those that would have beenavailable to domesticated animals in antiquity (Dudd & Evershed1998; Copley et al. 2003; Evershed et al. 2003). A large separation isobserved between the [[delta][sup.13.C.sub.18:0]] values and[[delta][sup.13]] [C.sub.16:0] values of carcass fats of pig andruminant origin ([approximately equal to] 7 [per thousand] and[approximately equal to] 4 [per thousand] respectively) allowing the twogroups to be distinguished with confidence. Compound-specific stable carbon isotope analysis has been usedextensively to distinguish between remnant fats of ruminant adipose ordairy origin, enabling questions regarding the origins of dairying inthe UK and elsewhere to be addressed (Copley et al. 2003; 2005a; 2005b;2005c; 2005d; Craig et al. 2005a ; 2005b; Spangenberg et al. 2006).Figure 2 shows how this technique can also distinguish archaeologicalporcine (pig, non-ruminant) from ruminam fats (e.g. cattle, sheep/goat),due to the different metabolic and digestive physiologies of these twogroups (Christie 1981; Enser 1991; Evershed et al. 1997; Mottram et al.1999). However, the potential of the latter aspect of the technique has,thus far, been little exploited (Dudd et al. 1999). We demonstrateherein the wider utility of the compound-specific approach toinvestigate the processing of pig products in prehistory prehistory,period of human evolution before writing was invented and records kept. The term was coined by Daniel Wilson in 1851. It is followed by protohistory, the period for which we have some records but must still rely largely on archaeological evidence to through organicresidues preserved in pottery residues. [FIGURE 2 OMITTED] Pigs in prehistory Domesticated do¡¤mes¡¤ti¡¤cate?tr.v. do¡¤mes¡¤ti¡¤cat¡¤ed, d o¡¤mes¡¤ti¡¤cat¡¤ing, do¡¤mes¡¤ti¡¤cates1. To cause to feel comfortable at home; make domestic.2. To adopt or make fit for domestic use or life.3. a. pigs in Britain are descended from the wild Sus scrofaL. found all over Europe, the Middle East and much of Asia. Thezooarchaeological record suggests pigs were first domesticated in theNear East c. 9000 years ago (Epstein & Bichard 1984), with molecularand archaeological evidence signifying a second independentdomestication domesticationProcess of hereditary reorganization of wild animals and plants into forms more accommodating to the interests of people. In its strictest sense, it refers to the initial stage of human mastery of wild animals and plants. in the Far East (Giuffra et al. 2000; Jing & Flad2002), However, a recent extensive genetic study has provided evidencethat there were multiple centres of pig domestication across Eurasia(Larson et al. 2005). Pigs would have been a valuable commodity toprehistoric pe ople; they have a short reproductive cycle reproductive cyclen.The cycle of physiological changes that begins with conception and extends through gestation and parturition. and largelitter size, thus, in favourable conditions they have an extraordinarycapacity to reproduce (Grigson 1982). Pigs eat virtually anything,thereby converting inedible organic debris to meat and in doing soproviding a means of controlling settlement waste (Gregg 1988). Despitethis, at the majority of Neolithic and Bronze Age sites in Britain,cattle bones dominate the animal bone assemblage (Pollard 1997; Darvill1998; Thomas 1999; Ray & Thomas 2003; Parker Pearson 2003). Previousstudies of lipid residues extracted from prehistoric British potteryhave shown relatively few vessels to contain predominantly porcinelipids (Dudd & Evershed 1998; Dudd et al. 1999; Copley et al. 2005a;2005b; 2005c; 2005d; Craig et al. 2005a; 2005b). This may be due toalternative methods of cooking pig meat, such as sp it roasting (e.g.Albarella & Serjeantson 2002), or it may be a dietary bias, wherebybeef, lamb and dairy products were cooked and consumed in preference topork. The high incidence of pig bones at some Late Neolithic GroovedWare sites makes this pottery ideal for testing the use of stable carbonisotope analysis to detect pig product processing. [FIGURE 3 OMITTED] Late Neolithic Grooved Ware pottery Grooved Ware pottery is tub-, bucket- or barrel-shaped, flat-basedand is characterised by grooved, incised and plastic decoration (Gibson1986). The tradition appears to have originated in northern Scotland,possibly as early as 3400 BC and certainly by 3100 BC (Ashmore 1998),and had spread into southern Britain by c. 2800 BC (Garwood 1999).Manufacture and deposition of Grooved Ware appears to have ceased by c.2000 BC, meaning the pottery style persisted for over 1000 years. LateNeolithic Grooved Ware pottery has been recovered at some sites inassociation with significant prop ortions of pig bones (e.g. Harcourt1971; Grigson 1981; Edwards & Home 1997; Hey et al. 2003) leading tospeculation that pigs were consumed during ceremonial feasts (e.g.Sherratt 1991; Richards & Thomas 1984; Albarella & Serjeantson2002; Hey et al. 2003). A preliminary study of absorbed and surfacelipid residues in Grooved Ware and the preceding Peterborough Ware, fromUpper Ninepence, Walton in the Welsh Borderlands demonstrated thatGrooved Ware vessels were used to process both ruminant and porcineproducts while Peterborough Ware vessels contained only ruminant fats(Dudd et al. 1999), a finding that seems to be supported by otherdata-groups such as the microwear analysis of stone tools at the site(Donahue 1999); however, too few vessels were analysed to drawdefinitive conclusions. Given its wide geographical distribution, long use period andpossible association with pig bones, Grooved Ware pottery appeared toprovide an ideal candidate for testing hypotheses rela ting to pigconsumption through compound-specific stable isotope stable isotopen.An isotope of an element that shows no tendency to undergo radioactive breakdown. determinations ofpreserved fats. Three major questions are addressed in this study: 1. Was pork (or other pig products) cooked in Grooved Ware vessels? 2. Was pork (or other pig products) cooked in Grooved Ware vesselsto a greater extent than in other prehistoric pottery traditions? 3. Can the incidences of porcine lipids in pottery be related tothe abundance of pig in the faunal assemblages? Selection of potsherds and analytical procedures Single potsherds from 385 vessels were selected from 16archaeological sites. Sites were chosen based on several criteria: (i)availability of potsherds; (ii) site type (including both domesticsettlements and 'ceremonial' sites); (iii) vessel type(including decorated and plain sherds, substyles and contemporaneousware types). Sites were also selected covering as wide a geograp hicalarea of the British Isles as possible. As fewer Grooved Ware sites havebeen discovered in the Midlands and North of England, sites were mainlyrestricted to southern England and Scotland and included: Links ofNoltland, Skara Brae, Stonehall and Crossiecrown (Orkney), BalfargRiding School (Fife), Melbourne (Lanarkshire), Yarnton Floodplain floodplain,level land along the course of a river formed by the deposition of sediment during periodic floods. Floodplains contain such features as levees, backswamps, delta plains, and oxbow lakes. (Oxfordshire), Betchworth (Surrey), West Kennet Palisade Enclosures andDurrington Walls (Wiltshire) and Wyke Down (Dorset). In addition toGrooved Ware, broadly contemporaneous pottery styles from the Neolithic(Round Based, Peterborough and Impressed Wares) and Bronze Age (Beakerand Flat Rim Wares) were also investigated from either the same sites orthose in close proximity to the selected Grooved Ware sites. Lipidconcentrations have been sho wn to be highest in rim or upper-body sherds(Charters et al. 1993), so these were selected wherever possible. The analytical procedures have been described in detail elsewhere(e.g. Evershed et al. 1990; Charters et al. 1993; Dudd & Evershed1998; Copley et al. 2003; 2005a). Briefly, 2g sub-samples of surfacecleaned potsherds were crushed and lipids extracted with organic solventto yield a total lipid extract (TLE TLE Temporal Lobe EpilepsyTLE The Living End (band)TLE Two Line Elements (for satellite tracking)TLE The Learning EquationTLE Taxe Locale d'¨¦quipement ). A portion of the TLE wastrimethylsilylated prior to gas chromatography gas chromatography (GC)Type of chromatography with a gas mixture as the mobile phase. In a packed column, the packing or solid support (held in a tube) serves as the stationary phase (vapour-phase chromatography, or VPC) or is coated with a liquid stationary phase (GC) and GC/massspectrometry (GC/MS GC/MS Gas Chromatograph/Mass Spectromete rGC/MS Gas Chromatograph/Mass SpectrometryGC/MS Gas Chromatograph/Mass Spectrograph ) screening, and a further aliquot aliquot(al-ee-kwoh) adj. a definite fractional share, usually applied when dividing and distributing a dead person's estate or trust assets. (See: share) saponified andderivatised to yield fatty acid fatty acid,any of the organic carboxylic acids present in fats and oils as esters of glycerol. Molecular weights of fatty acids vary over a wide range. The carbon skeleton of any fatty acid is unbranched. Some fatty acids are saturated, i.e. methyl esters (FAMEs) forGC-combustion-isotope ratio MS (GC-C-IRMS) analysis to determinecompound-specific [[delta][sup.13.C]] values of [C.sub.16:0] and[C.sub.18:0] fatty acids. Classification of fats in Grooved Ware and non-Grooved Ware pottery Grooved Ware (222) and non-Grooved Ware (163) potsherds wereanalysed as discussed above. GC and GC/MS confirmed significantconcentrations of lipid (>5 [micro]g [g.sup.-l]) in a substantialproportion of Grooved Ware vessels (58 per cent), while fewer of thenon-Grooved Ware vessels (36 per cent) contained lipid. All of theextracts were identified as degraded animal fats, though a small numberof vessels also showed evidence for degraded plant and insect waxes.Further classification of the fats rested on the [[delta][sup.13]] Cvalues of [C.sub.16:0] and [C.sub.18:0] fatty acids by comparison withthe values for reference values ref¡¤er¡¤ence valuespl.n.A set of laboratory test values obtained from an individual or from a group in a defined state of health. fats (Figure 3). As can be seen fromFigure 4 both ruminant and porcine carcass products, as well as ruminantdairy products were processed in the Grooved Ware vessels; many vesselsalso contained mixtures of these commodities, evidenced by the extractsplotting between the confidence ellipses for the reference fats. Inorder to determine the approximate proportion of porcine fats in thesemixtures we appl ied a mixing model utilising the percentage abundance ofeach fatty acid in flesh porcine and ruminant fats and their[delta.sup.13.C] values (Woodbury et al. 1995; Dudd & Evershed1998). Vessels with a contribution of porcine fat equal to or greaterthan 75 per cent of the total mixture were classed as'predominantly porcine'. This provides a measure of therelative 'importance of pig' in a given vessel- or site-typeenabling comparison of pig versus ruminant exploitation across differentvessel types and sites. Chi squared ([chi square chi square (kī),n a nonparametric statistic used with discrete data in the form of frequency count (nominal data) or percentages or proportions that can be reduced to frequencies. ]) tests for associationwere also applied to investigate whether any associations werestatistically significant. A second statistic, Cramer's V([upsilon up¡¤si¡¤lonor yp¡¤si¡¤lonn. Symbol The 20th letter of the Greek alphabet. ]) is calculated, to measure the st rength of an association andhas a value between 0 and 1, with values close to 1 indicating a strongrelationship (Fletcher & Lock 1991). On this basis 16 per cent ofGrooved Ware vessels contained significant proportions of porcine fatand half of these were used to process solely porcine products. Thenon-Grooved Ware vessels, while predominantly used for the processing ofdairy products, also showed evidence for the processing of porcineproducts, though only four (7 per cent) of the extracts were composed ofpredominantly porcine fats. [FIGURE 4 OMITTED] To put these results into a wider prehistoric context they wereincorporated into an accumulated dataset of [[delta].sup.13] C valuesfor pottery lipid residues from prehistoric Britain, which includesvalues for 167 Neolithic, 111 Bronze Age and 128 Iron Age potsherds(Copley et al. 2003; 2005a; 2005b; 2005c). This allowed us toinvestigate patterns of pig consumption/processing based on fats frompotsherds spanning the entire period of British prehistory. Residuescomposed of predominantly porcine lipids were found in 7 per cent ofNeolithic vessels (excluding Grooved Ware), 5 per cent of Bronze Agevessels and 0 per cent of Iron Age vessels; compared to 16 per cent ofthe Grooved Ware vessels. Furthermore, a statistically significantassociation was found between Grooved Ware pottery and the occurrence ofporcine lipids ([chi square] = 20.94, p < 0.0001, df = 1, [upsilon] =0.19), whereas no association was observed for the Neolithic(non-Grooved Ware), Bronze Age or Iron Age pottery. This finding is inaccordance with the available faunal data for Neolithic assemblages(e.g. Darvill 1998; Thomas 1999). Comparison of fats in Grooved Ware with faunal remains At sites where faunal information was available (Durrington Walls,Harcourt 1971; West Kennet, Edwards & Home 1997; Wyke Down, Legge1991; and Skara Brae, Clutton- Brock unpublished report), the proportionof pigs in the faunal assemblage was com pared with the number of vesselsfound to contain predominantly porcine lipid residues. A goodcorrelation ([R.sup.2] = 0.76) was observed. This suggests that at leastsome fat containing pork products (meat, fat, bone marrow, blood) fromthe majority of pigs at the site were processed or stored in potteryvessels. However, we must be cautious as the faunal data is a mixture ofMNI See Merom New Instructions. values and number of bones; a more accurate picture could beobtained with further faunal data and by calculating meat mass indices.This is, nevertheless, encouraging as it suggests that stable carbonisotope analysis of animal fat residues in archaeological pottery can beused to estimate pig numbers at sites where bones have not survived(e.g. due to taphonomic factors such as acidic soils). Conclusions This is the first study to specifically investigate pigexploitation using lipid residue analysis and, as such, has shown thatGC-C-IRMS is a valuable tool for the identificatio n of pork consumptionfrom lipid residues preserved in archaeological potsherds. We haveestablished that: 1. Pork was cooked in Grooved Ware vessels and 16 per cent ofGrooved Ware vessels were used solely or predominantly to process pigcarcass products (meat, fat, bone marrow or blood). Other vessels mayalso have contained small quantities of porcine lipids mixed with otheranimal fats. 2. Pork was cooked in Grooved Ware to a greater extent than inother prehistoric pottery wares. Grooved Ware vessels show astatistically significant association with porcine fats that is not seenin Neolithic (non-Grooved Ware), Bronze Age or Iron Age vessels. 3. Linear regression Linear regressionA statistical technique for fitting a straight line to a set of data points. analysis was used to compare the percentage ofpig bones (from MNI and number of bones) from four sites with thepercentage of vessels containing predominantly porcine lipids at thosesites. This demonstrated that the proportio n of pig lipids in pottery isa statistical reflection ([R.sup.2] = 0.76) of the proportion of pigbones in the faunal assemblages at Grooved Ware sites. Acknowledgements We would like to thank Ian Bull and Jim Carter, for technicalassistance and the Natural Environment Research Council (NERC NERC Natural Environment Research Council (UK)NERC North American Electric Reliability Corporation (Princeton, New Jersey, USA)NERC Northeast Recycling CouncilNERC National Environment Research Council ) forfunding the Bristol node of the Life Sciences Mass Spectrometry Facility(Agreement No. F14/6/13/01). The Wellcome Trust is thanked for providinga Bioarchaeology PhD studentship for A.J.M. (061666/Z/00/Z). NERC andEnglish Heritage are thanked for financial support for the workundertaken by M.S.C. and R.B. We also wish to thank the manyarchaeologists and museum curators who have provided potsherds makingthis work possible: Alison Sheridan, National Museums of Scotland;Rosamund Cleal, A lexander Keiller Museum; Andrew Deathe, Salisbury andSouth Wiltshire Museum; Gill Hey, Oxford Archaeology; Andrew Fitzpatrickand Rachael Seager Smith, Wessex Archaeology; Richard Jones, Universityof Glasgow The University of Glasgow (Scottish Gaelic: Oilthigh Ghlaschu, Latin: Universitas Glasguensis) was founded in 1451, in Glasgow, Scotland. ; Andrew Jones, University of Southampton In the most recent RAE assessment (2001), it has the only engineering faculty in the country to receive the highest rating (5*) across all disciplines.[3] According to The Times Higher Education Supplement ; Jonathan Cotton,David Williams and Martin Green. References ALBARELLA, U. & D. SERJEANTSON. 2002. A passion for pork: meatconsumption at the late Neolithic site of Durrington Walls, in P.Miracle & N. Milner (ed.) 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Evershed (1) * (1) Organic Geochemistry Unit, Bristol Biogeochemistry bi¡¤o¡¤ge¡¤o¡¤chem¡¤is¡¤try?n.The study of the relationship between the geochemistry of a region and the animal and plant life in that region.bi ResearchCentre, School of Chemistry, University of Bristol, Cantock'sClose, Bristol BS8 1 TS, UK (2) Department of Archaeological Sciences, University of Bradford The University of Bradford is a university in Bradford, West Yorkshire in the United Kingdom. HistoryThe university has its origins in the Bradford Schools of Weaving, Design and Building which in 1882 became the Bradford Technical College. ,Bradford, West Yorkshire BD7 1DP, UK * Author for correspondence (Email: r.p.evershed@bristol.ac.uk)