See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/223211246

Starch granules, dental calculus and new perspectives on ancient diet

Article  in  Journal of Archaeological Science · February 2009

DOI: 10.1016/j.jas.2008.09.015

CITATIONS READS

179 1,830

6 authors, including:

Karen Hardy Les Copeland

Autonomous University of Barcelona The University of Sydney
89 PUBLICATIONS   1,980 CITATIONS    199 PUBLICATIONS   8,547 CITATIONS   

SEE PROFILE SEE PROFILE

Matthew James Collins

University of Copenhagen
500 PUBLICATIONS   18,807 CITATIONS   

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Archaeometry in Heritage Science View project

ancient plant use View project

All content following this page was uploaded by Karen Hardy on 31 December 2018.

The user has requested enhancement of the downloaded file.

This article appeared in a journal published by Elsevier. The attached
copy is furnished to the author for internal non-commercial research
and education use, including for instruction at the authors institution
and sharing with colleagues.
Other uses, including reproduction and distribution, or selling or
licensing copies, or posting to personal, institutional or third party
websites are prohibited.
In most cases authors are permitted to post their version of the
article (e.g. in Word or Tex form) to their personal website or
institutional repository. Authors requiring further information
regarding Elsevier’s archiving and manuscript policies are
encouraged to visit:
http://www.elsevier.com/copyright
 Author's personal copy

Journal of Archaeological Science 36 (2009) 248–255

Contents lists available at ScienceDirect

Journal of Archaeological Science

journal homepage: http://www.elsevier.com/locate/jas

Starch granules, dental calculus and new perspectives on ancient diet

Karen Hardy a, *, Tony Blakeney b, Les Copeland c, Jennifer Kirkham d,
Richard Wrangham e, Matthew Collins a
a
BioArch, University of York, YO10 5YW, United Kingdom
b
Cereal Solutions, P.O. Box 201 North Ryde NSW 1670, Australia
c
Faculty of Agriculture, Food and Natural Resources, The University of Sydney, NSW 2006, Australia
d
Leeds Dental Institute, Clarendon Way, Leeds LS2 9LU, United Kingdom
e
Department of Anthropology, Harvard University, 11 Divinity Avenue, Cambridge, MA 01238, USA

a r t i c l e i n f o a b s t r a c t

Article history: Recent work in various parts of the world has suggested the possibility of ancient starch granules
Received 26 April 2008 surviving and adhering to archaeological artefacts. Often this information is used to infer aspects of diet.
Received in revised form 22 August 2008 One additional source for recovery of archaeological starch granules is dental calculus. The presence of
Accepted 5 September 2008
plant food debris in dental calculus is well known but has not been not widely investigated using
archaeological material. The extraction of starch granules from dental calculus represents a direct link to
Keywords:
the consumption of starchy food by humans or animals. Using dental calculus also sidesteps many other
Hunter gatherer diet
questions still inherent in using starch granules to reconstruct aspects of ancient diet, such as the effects
Carbohydrates
Starch degradation of diagenesis on their morphology; as the starches are trapped inside a concreted matrix they are less
Amylase digestion likely to alter over time. We used amylase digestion by a starch-specific enzyme to confirm the material
as starch.
Ó 2008 Elsevier Ltd. All rights reserved.

Starch-based foods constitute 50-70% of the energy intake of
most humans today (Atkins and Bowler, 2001). Evidence in the
form of a combination of archaeological remains and ethnographic
records suggests that starchy food also had an important role in
pre-agricultural human diet. But evidence for starchy foods such as
tubers, roots and seeds can be difficult to find on some archaeo-
logical sites and the recovery and analysis of starch granules found
in archaeological contexts have shed new light on far reaching
questions surrounding plant use and domestication (e.g. Aranguren
et al., 2007; Fullagar, 2006; Horrocks and Nunn, 2007; Perry et al.,
2006; Piperno et al., 2000, 2004; Samuel, 1996; Van Peer et al.,
2003).
Starches used for archaeological studies are extracted from
residual material that is found in places such as adhering to the
edges of flaked stone tools; as material that has accumulated in the
pores of the coarse granular structure of stones used for grinding;
inside pots and in sediment (Hardy, 2008a; Iriarte et al., 2004; Perry
et al., 2006; Piperno et al., 2000, 2004; Van Peer et al., 2003). But
this assumes the function of certain tool types; for example
grinding tools are assumed to have been used for food preparation.
While this is very probable in many cases, Baysal and Wright (2006)
have demonstrated that grinding tools may also have other non-
* Corresponding author.
E-mail address: karhardy@gmail.com (K. Hardy).
food-related uses. Similarly, with regard to flaked stone tools, while
some of these may have been used for collection and processing of
food, they had many other uses notably in relation to raw material
preparation (Hardy, in press-a; Hardy and Sillitoe, 2003).
The use of dental calculus to extract starch granules for dietary
reconstruction offers a direct link to consumption. The presence of
plant food debris in archaeological dental calculus has been known
about for some time (Cummings and Magennis, 1997; Dobney and
Brothwell, 1986, 1988; Gobetz and Bozarth, 2001; Hardy, in press-b;
Lieverse, 1999; Lilley et al., 1994) but has not been not widely
investigated. Assuming that things recovered from the mouth are
likely to have been consumed, (though there are exceptions, for
example inner bast fibres can be chewed to soften them, Hardy,
2008b), the extraction of starch granules from dental calculus
represents a direct link to the consumption of starchy food by
humans or animals.
Edible starchy plants can be found in most environments, for
example over 60 indigenous edible starchy plant species can be
found in Britain alone. Starch is a reserve polysaccharide of plants,
an end product of carbon fixation by photosynthesis. It is present in
most green plants and can be found in every type of plant tissue
including leaves, stems, roots fruit, seeds and even pollen grains. It
is the major carbohydrate and energy reserve in seeds and plant
tubers. Most cereal starch is located in the endosperm which is the
central and largest part of the grain, while starch granules are the
dominant component of tuberous root crops such as potatoes.

0305-4403/$ – see front matter Ó 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jas.2008.09.015
 Author's personal copy

K. Hardy et al. / Journal of Archaeological Science 36 (2009) 248–255 249

Starch is composed of a mixture of two glucose polymers – amylose

and amylopectin – which together form discrete and morphologi-
cally identifiable granules (Evers et al., 2004; Zobel, 1997). Starch
has a very low osmotic pressure and this provides plants with a way
of storing large reserves of glucose without influencing water
relations within their cells. Starch granules are relatively stable
structures, but they are also able to be readily decomposed to
water-soluble sugars by digestive enzymes (Radley, 1968).
Though starch is normally susceptible to enzymatic attack, it is
semicrystalline and can survive for long periods of time in a stable
environment, for example in micro-cracks in flaked and ground
stone tools, within fragments of plant cell walls or within dental
calculus.

1. Evidence of pre-Neolithic plant consumption

Understanding ancient diet is a key component of studies into

early prehistory even though there is often little direct surviving
evidence. Pre-Neolithic diet is studied using different techniques
including counts of bones and shellfish, stable isotope analysis, Fig. 1. Processing wild seeds Central Australia.
assessments of calorific values and residues in pots, but with a few
exceptions (Clark, 1976; Zvelebil, 1994), reconstructions of pre-
The domestication of plants and cultivation of crops is the
Neolithic diet tend to be based largely around meat or seafood with
control and management of starch sources. Examples include cereal
little attention paid to plant foods except hazelnuts. However, there
grains in the Near East, rice and millet in East Asia, maize and
is extensive evidence for starchy plants on pre-agricultural sites in
potatoes in the Americas and underground plant storage tissues in
Europe and the Near East (Aura et al., 2005; Hardy, 2007; Moore
SE Asia. The fact that starch-based foods were adopted so widely
et al., 2000; Weiss et al., 2004).
across the world as a primary food source suggests that human
We need plants to live (Hladik and Pasquet, 2002). Apart from
consumption of starch has a very long heritage.
the vitamins and minerals that plants provide, starch is a macro-
nutrient and an important source of carbohydrate for dietary
energy. Starch occurs as granules which are broken down into 2. Ancient starch analysis
glucose in the digestive system. Today dietary starch comes
mainly from foods such as bread, pasta, potatoes and rice. While Analysis of ancient plants from the remains of starch granules
the proportion of starchy foods may not have been as high in has been expanding rapidly as a research area in archaeology, but
a hunter gatherer society, a completely starch free diet is also a number of outstanding issues remain.
unlikely.
Humans have a basic metabolic need for high energy carbohy-
2.1. Starch identification
drates and it is very likely that starchy plants were important
possibly even as far back in evolutionary time as the ability to
Starch granules from archaeological contexts are identified on
exploit underground storage organs may be linked to the Last
the basis of the size and shape of granules, through the use of stains
Common Ancestor’s (LCA) ability to survive in savannah environ-
(Torrence and Barton, 2006) and by the birefringent Maltese cross
ments (Laden and Wrangham, 2005). Though a reduction in starch
pattern which is clearly visible under the light microscope in
intake is advocated in some modern diets, the long-term implica-
polarized light. Birefringence is a characteristic commonly found in
tions of an absence of starch in the diet may be negative, while the
materials with crystalline layers arranged in a concentric pattern.
amount of carbohydrate from animal tissue is too small to be
Together these tests give a good indication of whether a material is
considered quantitatively important (Garrow et al., 2000).
indeed starch, but none is sufficiently specific to confirm it.
One human group that has managed to survive with a low
The only unequivocal test for starch is to degrade it with an
carbohydrate input is the high Arctic population as their traditional
alpha amylase, an enzyme that is specific for the chemical linkages
diet was based almost exclusively on fish and seal products. They
contained in starch and that uniquely degrades starch. Pure prep-
would have replaced carbohydrates as their predominant dietary
arations of this enzyme may be obtained commercially; the
energy source with protein and fat. The negative physical conse-
currently favoured source for starch analysis is from Bacillus
quences of this diet, such as excessive urea production necessi-
licheniformis (see Section 3).
tating a high intake of water, mean that it is unlikely that this
would have been followed unless there was no alternative.
However the incorporation of large amounts of fat in the diet does 2.2. Cooking and gelatinization
suggest a way in which people could survive in periglacial envi-
ronments where few plants might have been available. Starch is insoluble in cold water but when heat is applied the
Ethnographic evidence indicates almost every known hunter granules go through a process called gelatinization. Temperatures of
gatherer group included plants in their diet. Percentages of plant gelatinization vary but gelatinization does not normally occur below
consumption for hunter gatherers range from 6–15% at high lati- 60  C. The rate and extent of gelatinization is influenced by the
tudes to 45–55% in tropical grasslands and desert scrub (Cordain application of shear forces (mixing, stirring) with heating. Gelati-
et al., 2000) (Fig 1). Additionally, there is a small but significant nization results in the disruption of molecular order, leading to
number of pre-Neolithic sites where charred remains of edible granular swelling and loss of birefringence caused by disruption of
tubers, seeds and legumes have been found (e.g. Aura et al., 2005; the semicrystalline structure. Retrogradation, the process in which
Clark, 1954; Kubiak-Martens, 1996, 1999, 2002; Mason and Hather, the starch chains begin to reassociate in an ordered structure, can
2000; Moore et al., 2000). occur once the gelatinized starch cools. This can result in a starch
 Author's personal copy

250 K. Hardy et al. / Journal of Archaeological Science 36 (2009) 248–255

which is more resistant to enzyme breakdown, and with greatly calculus as a source of granules rather than relying on artefacts and
altered morphological characteristics (van de Velde et al., 2002). sediments.
It still is not clear how starch granules and granule morphology Most people have dental plaque biofilms on and around their
persist in archaeological contexts (Torrence and Barton, 2006; teeth. Plaque biofilms comprise of complex mixed microbial
Haslam, 2004). Starch is readably biodegradable in the burial communities within an extracellular polysaccharide matrix.
environment, indeed it is a component of many biodegradable Calculus, which occurs when plaque biofilms accumulate and
plastics (Gordon et al., 2000; Imam et al., 1995, 1999). Perhaps mineralize, is associated with chronically poor oral hygiene (Little
retrograded starch granules which are resistant to biodegradation et al., 1963; Little and Hazen, 1964).
are selectively preserved? The rate of calculus formation is variable and is associated with
individual differences in diet, salivary flow, local pH and genetic
factors. Saliva is a rich source of amylase, which is produced by the
2.3. Granule morphology
salivary glands in addition to the pancreas. The starch granules in
uncooked or partially cooked starch need to be broken down for
If granule morphology changes during food processing (Angold,
digestion to occur and salivary amylase begins this process in the
1979), can it also change during diagenesis and if so would this
mouth. But some starch gets trapped in plaque before it has
compromise identification? At least in the case of bone, there
degraded. Once the starch is incorporated into the plaque it is
appear to be a number of parallels between cooking and diagenesis
protected from the salivary amylase.
(Roberts et al., 2002). Most starch granules range from 1 mm to
Saliva differs across species and among individuals in its
around 100 mm in size and their size, shape and morphological
chemical composition and mineral content and human dental
characteristics vary and can be used to identify the granule. Iden-
plaque biofilms differ at the mineral and microbial levels from one
tification is carried out using the typological concept of ‘best fit’.
person to another and between different ethnic groups (Roberts-
This method was used widely in earlier starch work (e.g. Reichart,
Harry et al., 2000). It is not clear to what extent differences in
1913, 1919) but is little used today by starch biochemists beyond
calculus among human groups is genetically determined or deter-
genus level identification. In archaeological reports, however,
mined by diet. However, it does not appear that these differences
granules are sometimes identified to species.
have an effect on the survival of starch in calculus. Calculus was
Much of the starchy material consumed would have been
common among non-industrialized communities in the past,
cooked before being ingested, raising questions as to how it
probably more due to the lack of dental hygiene than to genetic or
survived and how its origin could be identified. It is very likely that
dietary factors.
much open air cooking resulted in ungelatinised starch granules in
Dental calculus can be found around the teeth in the supra-
starchy food surviving the cooking process, particularly if there was
gingival area, which is above the gum-line, or in the subgingival
insufficient water or the material was not stirred adequately while
area, below the gum-line in the gingival crevice. Subgingival
heating. Recent ethnographic work has shown that open air
calculus in particular can accumulate and endure for long periods
roasting results in food that is not necessarily ‘cooked through’.
and may slowly build up throughout extended periods if it is not
Fig. 2 shows a cross-section of a bread (damper) that was cooked
removed (Ånerud et al., 1991). The gingival crevice is protected to
recently in a traditional open fire by an indigenous group in Central
a large extent from salivary amylase and may well therefore form
Australia. Only the exterior crust was indeed ‘cooked through’
an area of preferential survival of starch particularly as microbial
while the rest of the bread contained many unaltered starch
communities here are proteolytic rather than sacchrolytic (utilising
granules.
protein as substrates rather than sugars). The resulting metabolic
by-products of proteolytic metabolism, such as ammonia, result in
2.4. Starch in dental calculus localized raised pH. This in turn encourages plaque mineralization
as precipitation of calcium phosphate is favoured. Once calcified,
One final concern is the possibility of contamination by modern calculus is as hard as bone and is commonly preserved on
starch granules (Haslam, 2004; Mercader et al., 2007). Indeed the
presence of starch granules at a site or on an artefact does not prove
the use of plants for consumption. There has recently been renewed
interest (Boyadjian et al., 2007; Henry and Piperno, 2008) in dental

Fig. 3. Human mandible with subgingival calculus, Tarbat Medieval site, Portmaho-
Fig. 2. Internal structure of traditionally cooked bread, Central Australia. mack, Scotland.
 Author's personal copy

K. Hardy et al. / Journal of Archaeological Science 36 (2009) 248–255 251

archaeological teeth and is even found on some Pliocene hominid the dental calculus, each sample was placed in a small amount
teeth (Blumenschine et al., 2003). of 0.6 M HCl to determine whether it was indeed dental calculus, or
The aim of this paper is to analytically prove, for the first time, adhering sediment, in which case it quickly fizzes and dissolves.
the persistence of starch in the archaeological record and examine (This only occurred on one occasion). This process also serves to
the potential for using starch granules trapped in dental calculus to clean off any sediment adhering to the dental calculus. Following
gain insight into the consumption of starchy plants in prehistoric this, the calculus needs to be dissolved in order to extract the starch
diet (Fig. 3). and other plant material. Twenty milliliters 0.6 M, HCl was added to
each tube which were then rotated continuously at 4  C for 5 days.
The samples were mixed by vortexing and centrifuged at 3000g
3. Materials
for 15 min, washed, re-centrifuged in ultra pure water three times
and air dried. Once dry, samples were placed on microscope slides
Six samples each were taken from two recently deceased
and mounted in glycerol for viewing.
chimpanzees from the Kibale Chimpanzee Project, Uganda
(Carter et al., 2008). Small pieces of calculus, between 1 and 2 mm
long, were extracted from modern skeletal material in Kibale. To 4.2. Methods for amylase digestion
extract the calculus, skulls or jaws were held (using rubber gloves)
above a tray of silver foil. Teeth were scraped gently with a clean The method to degrade starch using thermostable B. lichen-
sharp knife. Grains or flakes of calculus that dropped onto the silver iformis a-amylase was proposed by Batey (1982) and refined by
foil were transferred directly into plastic tubes by forming the foil Hattey et al. (1994) before being developed into an analytical kit for
into a funnel and tapping it to direct the calculus residue into the total starch analysis described in McCleary et al. (1997) and
tube (Table 1). currently available as ‘‘A Total Starch Assay Kit’’ (Megazyme, 2006).
Samples were taken from three sets of human remains. Seven- To hydrolyse starch from archaeological samples the method had to
teen samples were taken from 16 human skeletons from the site of be modified for use with the minute samples normally obtained.
Kaman Kalehoyuk, Anatolia (Hardy, in press-b).Of these, 14 samples Samples of archaeological material were placed on microscope
contained starch granules. Calculus was also extracted from 6 slides, mounted in glycerol and sealed with high vacuum grease.
skeletons from Tarbat, a Pictish monastery site at Portmahomak, (Glycerol was chosen as a mounting medium as it is miscible with
northern Scotland (Carver, 2007). A further 2 samples were taken aqueous buffers but viscous enough to inhibit movement on the
from a skeleton from Roman York. slide.) Blakeney and Stone (1985) have used aqueous ethanol
buffers up to 80% to degrade starch with B. licheniformis a-amylase
and have shown (in unpublished studies) glycerol could also be
4. Methods
used.
A micropipette was used to inject the a-amylase under the cover
4.1. Calculus extraction method
slip and directly adjacent to a starch granule. The progress of the
enzyme digestion was followed photographically. First attempts
Where possible, samples of subgingival calculus were taken. The
used a heated stage directly on the microscope, however, the liquid
Kaman Kalehoyuk calculus was prized off the tooth using a dental
enzyme that was injected into the slide caused the starch to move
pick and caught in a clean sheet of paper and placed in plastic zip-
around the slide and the starches were repeatedly ‘lost’ while
top bags. Samples from Tarbat and Roman York were caught in
degradation occurred too quickly. In order to better control the
a Petri dish then placed in centrifuge tube. Before degradation of
speed of degradation and enable the starch to be located in time for
it to be imaged, the heating stage was removed and the slide was
heated for short intervals (2–3 min). Once the slide was removed
Table 1
from the heated stage, the degradation slowed down or stopped
Dental calculus samples from Kibale chimpanzees
and it became possible to maintain a better control over the
Individual Tooth Comments degradation process. This enabled the process to be photographed
Sebitoli 1 Lower left C Subgingival flake at different intervals during the degradation process.
Sebitoli 1 Lower left M3 A substantial flake of enamel, chipped off
accidentally
Sebitoli 1 Lower left C Tiny grains of black supragingival calculus 5. Results
Sebitoli 1 None Grains accumulated on silver foil from processing
above 3 samples, i.e. mostly from the lower left
5.1. Starch degradation
canine
Sebitoli 1 Lower right I2 Grains from scraping black tartar from valleys on
worn surface (supragingival) Starch granules from modern, medieval and ancient samples
Sebitoli 1 Lower right C Scraped from beyond gum-line (subgingival) were degraded using the method described. The speed of degra-
Sebitoli 1 Lower left C Scraping around gum-line, both beyond it and dation appeared to vary randomly, the fastest occurring in around
from exposed sections (sub- and supragingival)
30 min. Botanical source may be more important than starch age in
Sebitoli 1 Upper left M3 Calculus around gum-line, both beyond it and
from exposed sections (sub- and supragingival) determining the rate of degradation (Fig. 4).
KFB 18 Upper left M3 Calculus around gum-line, both beyond it and
from exposed sections; also a chip of enamel
(sub- and supragingival)
5.2. Chimpanzee calculus
KFB 18 Upper right M2 White detachable encrustation from beyond
gum-line on the outer side of the tooth Numerous starches were found to be present in a matrix, so far
(subgingival) unidentified, but likely to be plant material (Fig. 5). Grains consis-
KFB 18 Upper left PM1 Similar to above sample, easily detached
tent in size and morphology with starch granules were present.
(i.e. strictly PM3) encrustation (hopefully, calculus) from beyond
gum-line. Outer and inner sides of tooth. A chunk Some granules were between 35 and 50 mm in length and their
also came from the Upper left PM2 (subgingival) hilum (the growth centre for the concentric rings) was located at
KFB 18 Upper right C Less calculus visible than in those specimens, but one end of the elongated granule. Many (>100 in some samples)
it was not hard to remove. Outer and inner small (15–20 mm), angular granules were also present. Calculus
(subgingival)
from Sebitoli 1 contained both elongated and small, angular
 Author's personal copy

252 K. Hardy et al. / Journal of Archaeological Science 36 (2009) 248–255

Fig. 4. Degradation sequence, following injection of a-amylase, of ancient starch granule from Kaman Kalehoyuk, Turkey.

granules while calculus from KFB 18 contained only the small agriculture exists in Iron Age Central Anatolia, this may represent
angular granules. an example of wild foods collection and consumption.

5.4. Pictish Tarbat, Scotland

5.3. Iron Age Kaman Kalehöyük
A range of different shaped granules was found in the calculus
from an Iron Age skeleton from the site of Kaman Kalehöyük in
Anatolia. Fig. 6 shows a large elongated granule 80 mm long, with
the hilum at one end and an obvious extensive growth ring pattern
on the elongated surface. While some starches can be less easy to
identify, a large granule such as this one is most likely to have come
from a tuber because of its large size, elongated shape and smooth
sides, all of which suggest a tuber starch. As no record of tuber
Cereal starches, from wheat, rye, barley and triticale have
a bimodal distribution: that is they have two granule sizes, type A
and type B (Fig. 7a) which are unique to these cereals. At Tarbat,
based on the presence of charred barley grains, it is understood that
barley was cultivated (Hall, personal communication). Fig. 7b
demonstrates the bimodal distribution of a sample of starch from
dental calculus from this skeleton. Wheat and triticale are unlikely
to have been cultivated in northern Scotland at this time. Rye and
barley can sometimes be differentiated on the basis of the size of
 Author's personal copy

K. Hardy et al. / Journal of Archaeological Science 36 (2009) 248–255 253

Fig. 5. Starches in dental calculus from a chimpanzee skeleton, Kibale Research

Station, Uganda.

their type a granules, as rye are on average larger (10–35 mm)

compared to barley (10–25 mm with an average of 12–15 mm)
(Eliasson and Larsson, 1993). A sample of granules was measured
and the result falls within both the rye and barley parameters
(mean ¼ 20.33 mm, n ¼ 20). As barley (Hordeum vulgare) is the only
cereal to have been identified at this site, it is likely that the starch
in Fig. 7b is barley.

5.5. Roman York, England

Two partially degraded starches (Fig. 8) can be seen in the

image. Here the degradation is visible as cracking around the
outside of the starches and may be the result of impact such as
chewing.

6. Discussion
Fig. 7. Bimodal starches, (a) modern wheat, (b) from dental calculus, probably barley.

Structures that appear morphologically similar to starch gran-

ules have been used extensively in recent years to obtain far be produced by other material, even water bubbles. Likewise iodine
reaching interpretations on plant domestication and the use of stains can be used for starch identification, but the intensity of
plants. But the identification methods used, such as observation of staining varies with the type of starch, and some other poly-
a rotating Maltese cross visible under cross-polarized light, can also saccharides also stain. We argue that in order to use the material

Fig. 6. Starch from human dental calculus, Kaman Kalehoyuk, Turkey. Fig. 8. Damaged starches, Roman York.
 Author's personal copy
254 K. Hardy et al. / Journal of Archaeological Science 36 (2009) 248–255
thought to be starch granules found on archaeological sites, it is
essential to be sure of their identification first. The only way to do
this with complete confidence is degradation of a sample by
a starch specific enzyme such as alpha amylase.
While extraction of starches from archaeological tools under-
stood to be used for food processing can provide information on
possible dietary components, the interpretation is based on
assumed use of the artefacts. The use of starches trapped inside
dental calculus provides a direct link to food consumption. Using
dental calculus also sidesteps the other problems still inherent in
using starch granules to reconstruct aspects of ancient diet, as the
starches are trapped inside a concreted matrix they are unlikely to
be subject to diagenetic change. Though in most cases genus level
identification is possible, there is currently no biochemical method
that can identify starches to species level.
A range of samples of different ages and locations were tested.
Almost every sample of dental calculus that was observed con-
tained starch granules. When combined with recent reports from
other authors (Boyadjian et al., 2007; Henry and Piperno, 2008) this
suggests dental calculus is an excellent source of biochemically
unaltered starch granules. The next stage in this research is to
assess how representative of overall diet these starch granules are.
Acknowledgements
Hayfa Salman provided Fig. 7a. Prof. Don Brothwell is thanked
for providing samples of dental calculus from Tarbat, Dr. Sachichiro
Omura for providing samples of dental calculus from Kaman
Kalehöyük, Dr. Veronica Hunt for collecting these samples, Dr. Barry
McCleary (Megazyme International) for providing the pure alpha
amylase enzyme from Bacillus licheniformis. Thanks to Prof. Martin
Carver and Cecily Spall for access to analyses on Tarbat material by
Don Brothwell and Allan Hall in advance of publication, to Barbara
Tjikatu and family for collecting the grass seeds and cooking the
bread illustrated in Fig. 3 and to Judy Birmingham for her assis-
tance. Thanks also to Hannah Koon for her assistance in developing
the calculus extraction method. The chimpanzee samples were
made possible by the National Science Foundation (proposal
0416125 to R. Wrangham). Hardy was funded by an EU Marie Curie
Outgoing International Fellowship.
References
Ånerud, Å., Löe, H., Boysen, H., 1991. The natural history and clinical course of
calculus formation in man. Journal of Clinical Periodontology 18, 160–170.
Angold, R.E., 1979. Cereals and bakery products. In: Vaughn, J.G. (Ed.), Food
Microscopy. Academic Press London.
Aranguren, B., Becattini, R., Lippi, M.M., Revedin, A., 2007. Grinding flour in Upper
Palaeolithic Europe (25000 years bp). Antiquity 81 (314), 845–855.
Atkins, P.J., Bowler, I.R., 2001. Food in Society: Economy, Culture, Geography. Hodder
Arnold, London, New York.
Aura, J.E., Carrión, Y., Estrelles, E., Pérez Jordà, G., 2005. Plant economy of hunter–
gatherer groups at the end of the last Ice Age: plant macroremains from the
cave of Santa Maira (Alacant, Spain) ca. 12000–9000 b.p. Vegetation History and
Archaeobotany 14 (4), 542–550.
Batey, I.L., 1982. Starch analysis using thermostable alpha-amylases. Starch/Stärke
34, 125–128.
Baysal, A., Wright, K., Members of the Çatalhöyük Teams, 2006. Cooking, crafts and
curation: ground-stone artefacts from Çatalhöyük. In: Hodder, I. (Ed.), Changing
Materialities at Çatalhöyük: Reports from the 1995–1999 Seasons. Excavations
at Çatalhöyük, vol. 5, Monographs of the McDonald Institute for Archaeological
Research, University of Cambridge, British Institute for Archaeology at Ankara,
Cambridge and London, pp. 307–324.
Blakeney, A.B., Stone, B.A., 1985. Activity and action pattern of Bacillus licheniformis
alpha-amylase in aqueous ethanol. FEBS Letters 186, 229–232.
Blumenschine, R.J., Peters, C.R., Masao, F.T., Clarke, R.J., Deino, A.L., Hay, R.L.,
Swisher, C.C., Stanistreet, I.G., Ashley, G.M., McHenry, L.J., Sikes, N.E., van der
Merwe, N.J., Tactikos, J.C., Cushing, A.E., Deocampo, D.M., Njau, J.K., Ebert, J.I.,
2003. Late Pliocene Homo and hominid land use from Western Olduvai Gorge,
Tanzania. Science 299, 1217–1221.
Boyadjian, C.H.C., Eggers, S., Reinhard, K., 2007. Dental wash: a problematic method
for extracting microfossils form teeth. Journal of Archaeological Science 34,
1622–1628.
Carter, M.L., Pontzer, H., Wrangham, R.W., Kerbis Peterhans, J., 2008. Skeletal
pathology in Pan troglodytes schweinfurthiiin Kibale National Park, Uganda.
American Journal of Physical Anthropology AJPA, 135, 431–437.
Carver, M., 2007. Portmahomack. Monastery of the Picts. Edinburgh University
Press.
Clark, D.L., 1976. Mesolithic Europe: the economic basis. In: Sieveking, G.de G.,
Longworth, I.H., Wilson, K.E. (Eds.), Problems in Economic and Social Archae-
ology. Duckworth, London, pp. 449–481.
Clark, J.G.D., 1954. Excavations at Star Carr: an Early Mesolithic Site at Seamer, Near
Scarborough. Cambridge University Press, Yorkshire.
Cordain, L., Miller, J.B., Eaton, S.B., Mann, N., Holt, S.H.A., 2000. Plant–animal
subsistence ratios and macronutrient energy estimations in worldwide hunter–
gatherer diets. American Journal of Clinical Nutrition 71 (3), 682–692.
Cummings, L.S., Magennis, A., 1997. A phytolith and starch record of food and grit in
Mayan human tooth tartar. In: Pinilla, A., Juan-Tresserras, J., Machado, M.J.
(Eds.), Estado Actual de los Estudios de Fitolitos en Suelos y Plantas: The State-
of-the-Art of Phytoliths in Soils and Plants, Monografı́as del Centro de Ciencias
Medioambientales, CSIC, 4, Madrid, pp. 211–218.
Dobney, K., Brothwell, D., 1986. Dental calculus: its relevance to ancient diet and
oral ecology. In: Cruwys, E., Foley, R.A. (Eds.), Teeth and Anthropology. BAR
International Series, vol. 291. BAR, Oxford, pp. 55–81.
Dobney, K., Brothwell, D., 1988. A scanning electron microscope study of archaeo-
logical dental calculus. In: Olsen, S. (Ed.), Scanning Electron Microscopy in
Archaeology. BAR International Series, vol. 452. BAR, Oxford.
Eliasson, A.-C., Larsson, K., 1993. Cereals in Breadmaking: a Molecular Colloidal
Approach. CRC Press, USA.
Evers, A.D., Blakeney, A.B., Miller, C., 2004. A Short Course on Grain Morphology.
Cereal Chemistry Division, Royal Australian Chemistry Institute.
Fullagar, R., 2006. Starch grains, stone tools and modern hominin behaviour. In:
Ulm, S., Lilley, I. (Eds.), An Archaeological Life. Papers in Honour of Jay Hall.
Aboriginal and Torres Islander Studies Unit, The University of Brisbane.
Garrow, J.S., Philip, W., Trehearne, J., Ralph, A., 2000. Human Nutritional Dietetics.
Elsevier, Health Sciences.
Gobetz, K., Bozarth, S.R., 2001. Implications for Late Pleistocene mastodon diet from
opal phytoliths in tooth calculus. Quaternary Research 55, 115–122.
Gordon, S.H., Imam, S.H., Shogren, R.L., Govind, N.S., Greene, R.V., 2000. A semi-
empirical model for predicting biodegradation profiles of individual polymers
in starch-poly b-hydroxybutyrate-co-b-hydroxyvalerate bioplastic. Journal of
Applied Polymer Science 76 (12), 1767–1776.
Hardy, K. Recent stone tool use and material culture of the Wola, Papua New
Guinea. In: McCartan, S., Woodman, P., Schulting, R., Warren, G. (Eds.), Meso-
lithic Horizons: Papers Presented at the Seventh International Conference on
the Mesolithic in Europe, Belfast 2005. Oxbow Books, Oxford, in press-a.
Hardy, K. Survival, extraction and identification of starch granules at Kaman-
Kalehöyük, Turkey. In: AAS XVI. Kaman, Turkey, in press-b.
Hardy, K., 2007. Food for thought: starch in Mesolithic diet. Mesolithic Miscellany
18 (2), 2–11. Available from: http://www.york.ac.uk/depts/arch/Mesolithic/
mmpdf/18.2.pdf.
Hardy, K., 2008a. Starch. Catalhoyuk 2007 Archive Report Available from: http://
www.catalhoyuk.com.
Hardy, K., 2008b. Prehistoric string theory. How twisted fibres helped to shape the
world. Antiquity 82, 271–280.
Hardy, K., Sillitoe, P., 2003. Material perspectives: stone tool use and material
culture among the Wola, PNG. Internet Archaeology 14 Available from: http://
intarch.ac.uk/journal/issue14/hardy_index.html.
Haslam, M., 2004. The decomposition of starch grains in soils: implications for
archaeological residue analyses. Journal of Archaeological Science 31 (12),
1715–1734.
Hattey, J.A., Sabbe, W.E., Blakeney, A.B., Batten, G.D., 1994. Nitrogen and starch
analysis of cotton leaves using near infrared reflectance spectroscopy (NIRS).
Communications in Soil Science and Plant Analysis 25, 1855–1863.
Henry, A.G., Piperno, D.R., 2008. Using plant microfossils from dental calculus to
recover human diet: a case study from Tell al-Raqa’I, Syria. Journal of Archae-
ological Science 35, 1943–1950.
Hladik, C.M., Pasquet, P., 2002. The human adaptations to meat eating: a reap-
praisal. Human Evolution 17, 3–4.
Horrocks, M., Nunn, P., 2007. Evidence for introduced taro (Colocasia esculenta) and
lesser yam (Dioscorea esculenta) in Lapita-era (c. 3050–2500 cal. Yr BP) deposits
from Bourewa, southwest Viti Levu Islands, Fiji. Journal of Archaeological
Science 34, 739–748.
Imam, S.H., Gordon, S.H., Burgess-Cassler, A., Greene, R.V., 1995. Accessibility of
starch to enzymatic degradation in injection-molded starch–plastic composites.
Journal of Polymers and the Environment 3 (2).
Imam, S.H., Gordon, S.H., Shogren, R.L., Tosteson, T.R., Govind, N.S., Greene, R.V.,
1999. Degradation of starch-poly (b-hydroxybutyrate-co-b-hydroxyvalerate)
bioplastic in tropical coastal waters. Applied and Environmental Microbiology
65 (2), 431–437.
Iriarte, J., Holst, I., Marozzi, O., Listopad, C., Alonso, E., Rinderknecht, A., Montana, J.,
2004. Evidence for cultivar adoption and emerging complexity during the Mid-
Holocene in the La Plata Basin. Nature 432, 614–617.
Kubiak-Martens, L., 1996. Evidence for possible use of plant foods in Palaeolithic and
Mesolithic diet from the site of Ca1owanie in the central part of the Polish Plain.
Vegetation History and Archaeobotany 5 (1–2), 33–48.
 Author's personal copy
K. Hardy et al. / Journal of Archaeological Science 36 (2009) 248–255
Kubiak-Martens, L., 1999. The plant food component of the diet at the Late Meso-
lithic (Ertebolle) settlement at Tybrind Vig, Denmark. Vegetation History and
Archaeobotany 8 (1–2), 117–127.
Kubiak-Martens, L., 2002. New evidence for the use of root foods in pre-agrarian
subsistence recovered from the Late Mesolithic site at Halsskov, Denmark.
Vegetation History and Archaeobotany 11 (1–2), 23–32.
Laden, G., Wrangham, R.W., 2005. The rise of the hominids as an adaptive shift in
fallback foods: plant underground storage organs (USOs) and australopith
origins. Journal of Human Evolution 49, 482–498.
Lieverse, A., 1999. Diet and the aetiology of dental calculus. International Journal of
Osteoarchaeology 9, 219–232.
Lilley, J.M., Stroud, G., Brothwell, D.R., Williamson, M.H., 1994. The jewish burial
ground at Jewbury. In: The Archaeology of York, 12 (3). C.B.A., York, pp. 507–531.
Little, M.F., Hazen, S.P., 1964. Dental calculus composition. 2. Subgingival calculus:
ash, calcium, phosphorus, and sodium. Journal of Dental Research 43, 645–651.
Little, M.F., Casciani, C.A., Rowley, J., 1963. Dental calculus composition. 1. Supra-
gingival calculus: ash, calcium, phosphorus, sodium, and density. Journal of
Dental Research 42, 78–86.
Mason, S., Hather, J., 2000. Parenchymatous plant remains. In: Mithen, S. (Ed.),
Hunter–gatherer Landscape Archaeology: the Southern Hebrides Mesolithic
Project 1988–1998. McDonald Institute Monographs, pp. 415–425.
Megazyme, 2006. Total Starch Assay Procedure (Amyloglucosidase/a-Amylase
Method). Megazyme International Ireland Limited.
Mercader, J., Barton, H., Gillespie, J., Harris, J., Kuhn, S., Tyler, R., Boesch, C., 27 February
2007. 4300-Year-old chimpanzee sites and the origins of percussive stone tech-
nology. Proceedings of the National Academy of Sciences 104 (9), 3043–3048.
McCleary, B.V., Gibson, T.S., Mugford, D.C., 1997. Measurement of total starch in
cereal products by amyloglucosidase – a-amylase method: collaborative study.
Journal of AOAC International 80, 571–579.
Moore, A.M.T., Hillman, G., Legge, A.J., 2000. Village on the Euphrates. From
Foraging to Farming at Abu Hureyra. Oxford University Press.
Perry, L., Sandweiss, D.H., Piperno, D.R., Rademaker, K., Malpass, M.A., Umire, A., de
la Vera, P., 2006. Early maize agriculture and interzonal interaction in southern
Peru. Nature 440, 76–79.
View publication stats
255
Piperno, D.R., Ranere, A.J., Holst, I., Hansell, P., 2000. Starch grains reveal early root
crop horticulture in the Panamanian tropical forest. Nature 407, 894–897.
Piperno, D.R., Weiss, E., Holst, I., Nadel, D., 2004. Processing of wild cereal grains
in the Upper Palaeolithic revealed by starch grain analysis. Nature 430,
670–673.
Radley, J.A., 1968. The a-amylases. In: Radley, J.A. (Ed.), Starch and its Derivatives,
fourth ed. Chapman and Hall, London.
Reichart, E.T., 1913. The Differentiation and Specificity of Starches in Relation to
Genera, Species, Etc., vol. 1. Washington Carnegie Institute.
Reichart, E.T., 1919. The Differentiation and Specificity of Starches in Relation to
Genera, Species, Etc., vol. 2. Washington Carnegie Institute.
Roberts, S.J., Smith, C.I., Millard, A., Collins, M.J., 2002. The taphonomy of cooked
bone: characterizing boiling and its physico-chemical effects. Archaeometry 44
(3), 485–494.
Roberts-Harry, E.A., Clerehugh, V., Shore, R.C., Kirkham, J., Robinson, C., 2000.
Morphology and elemental composition of subgingival calculus in two ethnic
groups. Journal of Periodontology 71 (9), 1401–1411.
Samuel, D., 1996. Investigation of ancient Egyptian baking and brewing methods by
correlative microscopy. Science 273, 4888–4890.
Torrence, R., Barton, H., 2006. Ancient Starch Research. Left Coast Press, California.
van de Velde, F., van Riel, J., Tromp, R.H., 2002. Visualisation of starch granule
morphologies using confocal scanning laser microscopy (CSLM). Journal of the
Science of Food and Agriculture 82 (13), 1528–1536.
Van Peer, P., Fullagar, R., Stokes, S., Bailey, R.M., Moeyersons, J., Steenhoudt, F.,
Geerts, A., Vanderbeken, T., De Dapper, M., Geus, F., 2003. The Early to Middle
Stone Age transition and the emergence of modern human behaviour at site 8-
B-11. Sai Island, Sudan. Journal of Human Evolution 45, 187–193.
Weiss, E., Kislev, M.E., Simchoni, O., Nadel, D., 2004. Small-grained wild grasses as
staple food at the 23 000 year-old site of Ohalo II, Israel. Economic Botany 58
(Suppl.), s125–s134.
Zobel, H.F., 1997. Molecules to granules: a comprehensive starch review. Starch/
Stärke 40, 44–50.
Zvelebil, M., 1994. Plant use in the Mesolithic and its role in the transition to
farming. Proceedings of the Prehistoric Society 60, 35–74.