Microbial Taphonomy of Archaeological Bone

Author(s): A. M. Child
Reviewed work(s):
Source: Studies in Conservation, Vol. 40, No. 1 (Feb., 1995), pp. 19-30
Published by: International Institute for Conservation of Historic and Artistic Works
Stable URL: http://www.jstor.org/stable/1506608 .
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MICROBIALTAPHONOMY OF ARCHAEOLOGICALBONE
A.M. Child
Summary-Taphonomy is the study of all changes that occur within an animal or plant following death. Bone
is the predominant material of animal origin to survive within the archaeological environment.It is the source
of a wealth of information concerning the relationships, diets, disease and ages of people (and animals) in
past cultures. For this information to be extracted and interpreted, there is a need for conservators and
archaeological scientists to be aware of the processes of taphonomic change which may occur in bone. These
changes are legion; this paper attempts to define some of the major changes seen in bone which may be attributable to the various actions of microorganisms.

Introduction
Taphonomy is the study of all changes occurring
within a substrate following death. These changes
are many and varied, and it is not the aim of this
article to present a comprehensive list. The main
factors involved in the taphonomic changes of bone
in the burial environment will be discussed below.
Bone is the predominant material of animal origin to survive within the archaeological environment. It is a composite material, comprising both
inorganic and organic fractions. Its study reveals
considerable information to the conservator, the
bone specialist and the archaeological scientist. For
this information to be retrieved, the conservator
must be aware of the likely mechanisms of decomposition and the state of preservation of the excavated bone.
One of the major problems confronting analysts
and conservators of archaeological bone is the
assessment of its integrity, since conservation treatments will be dictated by degree of preservation.
Both the inorganic and the organic phases of bone
can be changed by taphonomic processes, and estimation of the degree of preservation of these phases
based upon external morphology has been proved
to be unreliable [1].
Definitions
For clarity, some of the terms used in this paper
have been defined here. Other definitions exist,
however; the definitions given here have been
selected because they represent the most common
usage.
Biodegradation: any change in the properties of a
material caused by the activities of organisms.

Biodeterioration: any undesirable change in the

properties of a material caused by the activities of
organisms [2].
Biostratinomy: changes occurring in an animal or
plant after death but before burial [3].
Diagenesis: changes occurring in the animal or
plant following death and burial [3].
Microbial decomposition: deleterious changes to a
substrate due to the action of microorganisms, their
metabolic by-products and their enzymes.

Bone structure
Bone is a highly specialized composite material
comprising both inorganic (mineral) and organic
(mostly protein) phases. Its inorganic fraction (90%)
is calcium hydroxyapatite. The remaining 10% is
organic, made up of collagen, non-collagenous proteins (NCP), lipids, mucopolysaccharides and other
carbohydrates [4]. There are two types of bone,
cancellous bone (spongy bone) and compact bone
(cortical bone), and their distribution within the
body is dictated by biomechanical considerations.
Both compact and cancellous bone are formed by
the deposition of collagen fibrils in layers. In compact bone, these layers are laid down in rings
around the osteone, their alignment changing by
approximately 90? in each layer [5].
Bone mineral

Received 30 June 1993

Received in revisedform 22 August 1994

Hydroxyapatite, Ca10(PO4)6(OH)2,is the basis of

the inorganic component of bone. Bone mineral is a
complex substance, not stoichiometric with respect
to hydroxyapatite. Its calcium to phosphorus ratio
is significantly less than 10:6, so that the apatite
requires, in addition to its calcium, phosphate and
hydroxy ions, substantial amounts of carbonate and

Studies in Conservation40 (1995) 19-30

19

A.M. Child
lesser quantities of pyrophosphate, magnesium,
sodium and potassium [6]. Strontium and lead,
ingested as part of the diet of an individual, are
stored in the skeleton. Fluoride ions have a high
affinity for bone mineral, converting hydroxyapatite
to fluorapatite, and fluoride analyses have been
used to check provenance [7].

Bone proteins
The organic fraction of bone comprises 10-15% of
the total bone weight. There is one major protein
component, collagen, and a group of proteins
termed collectively non-collagenous protein (NCP).

Collagen

The collagen triple-helix is present in abundance in

a range of different vertebrate tissues. Thirteen different collagen types have been described, varying
in their chain composition and amino acid sequence
[8]. The collagen present in bone is Type I, which
comprises two chains of one composition, al(I),
and one of another, a2(I). Bone collagen has an
average composition of 30% glycine, 14% proline
and 11% hydroxyproline, the remainder making up
less than half the total amino acids [9]. Collagen is
the only human protein in which hydroxyproline
occurs in significant amounts; this amino acid
restricts the action of proteases [10]. In the Type I
collagen molecule, the protein chains are approximately 1000 amino acids (residues) long with
glycine (the smallest amino acid) occurring every
third residue to give the dominant sequence:
glycine-proline-hydroxyproline
Because of the presence of glycine residues, the collagen chains are tightly twisted; chains which are
tightly twisted have an increased resistance to the
action of proteolytic enzymes. The chains contain
hydrogen bonds between the amide nitrogen of
glycine in one chain and the non-glycine carboxyl
oxygen in an adjacent chain [11]. Mature Type I
collagen is further strengthened by a covalent bond
which forms between lysyl amino acid [12]. This
covalent bond is peculiar to bone collagen, and
makes the molecule even more resistant to enzymatic cleavage and laboratory extraction procedures. The presence of the mineral, which is in
intimate association with the collagen, further
inhibits the access and action of enzymes.
Non-collagenousproteins
Non-collagenous proteins (NCP) are a complex
group. Bone contains two sources of NCP, one
arising from outside the bone (i.e., a-2HS-glycoprotein and albumin) and the other from the bone
itself (i.e., osteonectin and osteocalcin). Osteonectin

20

is thought to be involved in the deposition and

development of primary bone tissue, and osteocalcin is concerned with the remodelling and development of secondary bone tissue. The bone-specific
NCPs are high in aspartic and ycarboxyglutamic
acid residues [13].

Bone as an archaeological resource

As stated above, bone is the predominant material
of animal origin to survive within the archaeological environment. Its study reveals considerable
information to the conservator, the bone specialist
and the archaeological scientist. Archaeological
skeletal materials provide two avenues of investigation: palaeopathology and archaeological science.

Palaeopathology
The gross morphology of archaeological bone can
be changed by the burial environment, but physical
measurements of skeletal bone dimensions will still
yield information on growth, health and incidence
of some diseases within human populations. Similar
examination of animal bone produces information
relating to butchering methods, diet, hunting and/or
farming techniques practised by ancient populations
[14]. Microscopic examination of archaeological
bone (and the related surviving soft tissues) will
yield more information on disease [15].

Archaeological science
The preservation of bone depends not upon the
length of burial but upon the environment of burial. If bones are sufficiently well-preserved, they still
contain indigenous organic macromolecules. These
can be used to provide information on:
-the age of the bone by radiocarbon dating
[16-18] or by amino acid racemization [19-22];
-the diet of the animal by stable isotope analysis
[23-26];
-genetic relationships [27-29].
This information is only reliable where the proteins
used have survived unchanged by biological, physical or chemical degradative processes [30].

Survival of proteins in archaeological bone

This article is restricted to a discussion of the
effects on bone of the microbes within terrestrial
environments. It is assumed here that the bone
(with or without flesh) enters the burial environment in direct contact with the soil, and is not
buffered by the presence of a coffin, shroud or
other protective layer.

40 (1995) 19-30
Studiesin Conservation

Microbialtaphonomyof archaeologicalbone
Bone protein is unusual in that it survives over
archaeological, even geological, time scales.
Generally, protein loss from unmineralized substrates is considered to be exponential, so that the
amount which survives depends upon the initial
concentration [31]. The protein loss within archaeological bone does not follow this pattern and, as a
result, archaeological bone can contain only 1-2%
nitrogen [26]. Indeed, the carbon to nitrogen ratio
in archaeological bone collagen does not change
from that of modem bone until almost all of the
collagen (97%) has been lost [23]. Where 98% of
the original bone protein has been lost, the amino
acid content is enriched in aspartic and glutamic
acid residues [26, 32]. The original source of these
residues is uncertain; however, three possibilities
exist:
-contamination of the bone proteins with soil proteins [33];
-survival of the NCP, rich in these residues, whose
presence is masked when collagen survives well
[32];
-selective decomposition of the collagen helix, to
leave acidic amino acids of collagen origin.
The degree to which proteins survive intact in
bones from archaeological contexts can be measured using immunochemical means. Although
these methods are well established for application
to modem materials, they have only recently been
applied to archaeological ones [34-39]. Results must
be viewed with caution [40]; however, they show
that indigenous proteins (i.e., apohaemoglobin and
osteocalcin) survive in an immunologically recognizable form in bones.
Proteins (both collagens and NCP) are thought
to survive in bone because of their close association
with the mineral phase, which protects against
enzyme attack. The collagens are further preserved
because of their innately protective structure and
chemistry (see 'Collagen' above).

Dissolution of the bone mineral

The response of the mineral and organic phases of
bone to the burial environment will differ. Janaway
[41] suggested that decomposition of bone probably
occurs in the later stages of the decomposition of a
corpse, but this is not likely. Deterioration, both
biochemical (autolysis) and microbiological, of tissue begins immediately after death, the soft tissues
usually showing the effects first. If the microbiological decomposition of bone is inhibited by the continued presence of the mineral phase, then the
degradation of collagen will proceed by chemical
means (see below).

40 (1995) 19-30
Studiesin Conservation

Dissolution due to the soil chemistry

The chemicaland physical deteriorationof buried
bone depends solely upon the chemistry(and biochemistry)of the surroundingburial environment.
Acidic soils will dissolve hydroxyapatite,but the
threshold of acidity requiredis not clear; at pH
< 5, demineralizationis promoted[42]. The rate of
dissolutionwill depend on the pH of the soil, the
concentrationof chelatingagents and the degreeof
water percolation.Bone tends to survivebetter in
soils with a neutral or very slightly alkaline pH.
This is because the bone mineral is less likely to
dissolve, but it must be noted that soils low in
phosphatewill also promotedemineralization[43].
Calcium hydroxyapatite, Ca10(PO4)6(OH)2, can

be changedboth by dissolutionand recrystallization

and by hetero-ionic substitution (e.g. Fe2+, Ca2+).

Vivianite (Fe3(PO4)2 8H20), brushite (CaHPO4) and

calcite (CaCO3) will form within archaeological

bones dependingupon the pH and chemistryof the
soil matrix. Soils high in Ca2+will lead to the
break-upof the bone, because CaCO3,formed in
high-calciumsoils, occupiesa largerspace than the
crystals of Ca10(PO4)6(OH)2. The presence of CaCO3

exerts internal stresseswithin the bone which will

result in embrittlement and cracking. Internal
stresseswhich cause damageto the gross morphology of bone are inducedwhen bone is exposed to
alternating wetting/drying and freezing/thawing.
These stresses are aggravatedby the presence of
solublesalts [44].
Dissolution due to microbial decomposition
Generally,in well-aeratedsoils, the decomposition
of organic matter is rapid. The high mineralcontent of bone, however, will initially inhibit the
microbialdecompositionof collagen. This decomposition will be affected by the relative numbers
constituting the microbial population of different
soils. The composition of the soil microflora is
highly dependentupon soil pH, and the metabolic
activityof microorganismswill influencethe pH of
the soil, especially those activities that involve
redox reactions [45]. The conditions which affect
the survivalof bone in the archaeologicalenvironment [46] are also those which will affect the
growth and survivalof microorganismsin the soil
[45, 47].
Microorganismswill penetrate into bone using
naturalspaces(e.g. blood vessel and nervelacunae),
releasingtheir enzymesand depositingamino acids
[48, 49]. They will grow using the surrounding
unmineralizedtissues as a food source and excrete
their secondary metabolites. The nature of these
productsof microbialmetabolismwill dependupon
21

A.M. Child
the environment in which the decomposition
occurred.
Anaerobic and aerobic decomposition
Microorganisms can be divided into three categories, depending upon their metabolism: they may
be either obligately aerobic, facultatively anaerobic
or obligately anaerobic [50].
In aerobic environments, complete decomposition
of the protein to carbon dioxide, water, nitrogen
dioxide and sulphur dioxide occurs without a considerable reduction in pH; therefore the proteins in
bone are more likely to survive. All the fungi and
some of the bacteria have an obligately aerobic
metabolism, degrading complex proteins with proteolytic enzymes. Rapid aerobic growth will promote
anaerobic conditions; this occurs when the rate of
oxygen consumption exceeds the rate of oxygen diffusion into the system. When anaerobic conditions
have been achieved, the obligate aerobic microorganisms will stop growing, but the bacteria that
have the facility for both anaerobic and aerobic
decomposition will switch to anaerobic pathways.
The obligate anaerobes will start to flourish.
Anaerobic fermentation uses substances other
than molecular oxygen as a terminal electron acceptor [50]. Microbial decomposition of protein under
anaerobic conditions leads, initially, to a drop in
pH. This is due to the generation of acidic compounds such as low molecular weight fatty acids
and amino acids, which are by-products of anaerobic metabolism. Their presence will result in the
demineralization of hydroxyapatite and the exposure of collagen to collagenolytic enzymes. Extrapolating from work on dental caries [51], an
increase in proton concentration will result in the
partial demineralization of hydroxyapatite, because
it will buffer the changes in pH at its own expense.
Microbial fermentation of bone proteins by
anaerobic pathways could lead to the total loss of
the organic fraction of bone since the products of
the reaction perpetuate the reaction; total loss of
the organic fraction, however, is not likely. As the
rate of anaerobic decomposition slows due to the
lack of metabolites, oxygen will diffuse into the system. Eventually, the rate of anaerobic degradation
will be surpassed by the rate of oxygen diffusion
and the system will return to its aerobic state, when
the rate of aerobic degradation will be determined
by the concentration of bio-available organic matter [52].
Catalyzing bone decomposition
To decompose bone, a microorganism must be able
to obtain energy from the collagen and mineral.
22

For the microbial enzymes to gain access to the collagen, the microbe must also be able to demineralize the bone, or grow in an environment where
demineralization occurs (e.g. a low-pH soil).
Enzymes
Enzymes are proteins which can increase the rate of
(catalyze) biological reactions between 106- and 108fold. As well as increasing rate, enzymes are specific
in the reactions which they will affect. An individual enzyme will catalyze a specific reaction with a
unique substrate or group of substrates. Enzymes
that disrupt proteins are called proteolytic enzymes
or proteases, and enzymes that are unique in their
ability to hydrolyze efficiently the triple-helical
regions of collagen under physiological conditions
(i.e., moderate temperature and around neutral pH)
are called collagenases. Collagenases are complexes
of several different enzymes, each of which has a
different catalytic function. There are two accepted
types of collagenase [53].
Collagen degradation by vertebrate collagenases
Vertebrate collagenases (tissue collagenases) disrupt
unmineralized collagen into only two fragments by
action at a specific site within the al(I)-chain, a single glycine-isoleucine bond between residues 772
and 773 [54]. The enzyme appears, therefore, to recognize the whole collagen molecule configuration.
Collagen degradation by microbial collagenases
On the other hand, microbial collagenases recognize
small amino acid sequences. All microbial collagenases examined so far appear to have the same
amino acid sequence requirements for cleavage
[55-59]; hence examination of the action of a wellcharacterized bacterial collagenase can be included
here.
The anaerobic bacterium Clostridiumhistolyticum
produces a collagenase complex of six enzymes. All
are highly active against collagen and devoid of
other proteolytic activities. Study of their amino
acid sequence requirements for cleavage is challenging, since purification of these enzymes is difficult;
however, the requirement for one of them (,3-collagenase) has been demonstrated [60]. Unlike vertebrate collagenases, 3-collagenase does not have a
rigid sequence requirement, but will cleave between
the X- and the glycine- residues in any of the
sequences below:
-glycine-Y-X-glycine-proline-hydroxyproline-glycine-Y-X-glycine-alanine-arginine-glycine-Y-X-glycine-Z-alaninewhere X-, Y- and Z- can be any of the amino acids
which constitute collagen, except glycine. Microbial
Studies in Conservation40 (1995) 19-30

Microbial taphonomy of archaeological bone

collagenases, therefore, have active sites along the
length of the collagen molecule which are dictated
by amino acid sequence rather than whole molecule
configuration. Like tissue collagenases, they are
metalloproteinases containing a zinc ion at the
active site [61, 62].
Microbial collagenases cleave the collagen helix
into a range of short peptides by hydrolysis at multiple sites along the triple helix. The major product
following treatment of Type I collagen with the collagenase complex from Cl. histolyticum is the
tripeptide glycine-proline-hydroxyproline, although
many other products are generated. Like vertebrate
collagenases, microbial collagenases require that the
collagen triple-helix be demineralized before the
enzyme can gain access [63].
Collagen degradation by proteases
Microorganisms that produce collagenase may vary
in the rate at which they will decompose the bone.
The microbial by-products of metabolism and the
presence of other enzymes (e.g. proteases) can also
affect the collagen. Proteases appear to have no
action on the collagen triple-helix; it could be
argued that their only value in collagen decomposition is that of reducing the short peptides released
by collagenase action to single amino acids. It was
suggested above (see 'Collagen degradation by
microbial collagenases') that the only enzyme system capable of cleaving the chains of Type I collagen in their helical regions is the collagenase
system. This may not be the case.
One protease, chymotrypsin (enzyme classification number: E.C. 3.4.21.1), has been shown to
have collagenolytic activity.
Classically, chymotrypsin should have little effect upon collagen,
since peptide bonds which involve hydroxyproline
are resistant to its action [10]. Chymotrypsins from
the mid-gut of shrimps (Penaeus monodon, P. japonicus and P. penicillatus) have been shown to have
some collagenolytic activity [64]. The shrimp chymotrypsins are resistant to the standard a-chymotrypsin inhibitors, and some have been shown to
have a higher affinity for collagenolytic activity
than for other proteolytic activities [65].
Collagenolysis by non-specific proteases has been
promoted by low pH [66]. Pepsin (E.C. 3.4.23.1) in
acetic acid is the standard laboratory method by
which Type I collagen is removed from demineralized bone [67]. This enzyme is used to cleave the
covalent bonds holding the triple-helix together.
Unlike collagenase, it has no action upon the
helices themselves. Indeed, low pH could be the
mechanism by which some fungi produce 'tunnels'
(see 'Diagenetic changes' below) within archaeological bone [68].
Studies in Conservation40 (1995) 19-30

Although no microbial proteases capable of

cleaving collagen in its helical parts have yet been
described, the possibility of this decomposition
pathway cannot be ruled out.

The taphonomy of bone

Taphonomy was defined above as the study of all
changes which occur within an animal after death
[3]. The taphonomy of bone is influenced by the
physical and chemical characteristics of the surrounding environment and the destruction of the
surrounding soft tissue. There are two main
processes by which bones and soft tissue become
degraded, both of which affect the stability of the
associated collagen. These processes are enzymatic
(which includes both autolysis and microbial
decomposition) and chemical; both are affected by
temperature.
Limitations of burial temperature
The range of soil temperatures is dictated by the air
temperature and the soil depth. In Britain, at a
depth of 300cm, the temperature range is 10.5 ?
2?C [69]. Published studies of the action and characteristics of microbial collagenase have been carried out at 28?C and 37?C. The microorganisms
that will produce collagenases at soil burial temperatures cannot be extrapolated from this literature,
since a significant temperature reduction of 1827?C will severely restrict many different species,
including Clostridiumhistolyticum [70].
Microbial studies at low temperatures are
required. Although only a few studies of this
nature have been undertaken, results so far indicate
that certain types of bacteria and fungi can be isolated repeatedly. The bacteria and fungi able to
produce collagenases at low temperatures are present in archaeological bone and associated soils in
very low numbers when compared to the total
microbial population. The bacteria and fungi are
obligate aerobes, able to grow over a wide range of
temperatures (4-39?C) and pH values (3-6-9-0) [68,
71]. The bacterial species isolated (Pseudomonas,
Aeromonas, Xanthomonas) produce metabolites
which are toxic to fungi and therefore can compete
successfully with a wide range of fungal species (see
'Microbial interactions' below).
Autolytic changes
As a cell dies, it releases a mixture of enzymes from
the lysosomes. These enzymes have an autolytic
(self-destructive) function and usually take the form
of proteinases and DNAses, which speed up the
destruction of the tissue and its component cells.

23

A.M. Child
Autolysis usually follows very rapidly after death;
even in the short interval between death and burial,
the osteocytes, marrow and neuro-vascular bundles
undergo autolysis [72]. Autolysis is self-limiting
and, after a certain stage is reached, the cells
remain stable for long periods of time [73]. The
majority of changes in bone, however, affect the
extracellular matrix proteins (i.e., collagen) which
are not amenable to autolysis.
Microbial changes
Augmenting the processes of autolysis is microbial
decomposition: autolysis opens up the soft tissues,
thus increasing access for microorganisms.
Decomposition of whole bodies involves the loss of
both soft and hard tissues, and both aerobic and
anaerobic environments will be achieved within the
rotting flesh.
Due to the presence and high numbers of gut
flora (which include some microorganisms capable
of inducing dental caries [74]), the bones within the
abdomen and thorax will suffer the demineralizing
effects of putrefaction for longer than the long
bones or the skull. This is borne out in some studies, but not in others [75, 76]. Laboratory studies
using bones inoculated with microorganisms [77]
support the premise that soft tissue destruction augments hard tissue loss.
Biostratinomic changes
Autolytic and microbial changes appear very soon
after death. Autolytic destruction can occur within
10 seconds following death of the cell [73]. The
types and degree of microbial destruction will be
dictated by the environment in which the corpse
lies. If burial does not immediately follow death,
the growth of microorganisms which would normally be prohibited by the low burial temperatures
will be promoted (e.g. Cl. histolyticum). Once the
body has cooled, these microorganisms are unlikely
to grow, but it is possible that the temperature of
the decaying body may increase sufficiently to allow
for their growth.
It is probable that Cl. histolyticum has its most
important role in the breakdown of collagen in the
pre-burial stage. Collagen breakdown will occur as
the bones and associated tissues lie on (or slightly
below) the surface of the soil. Cl. histolyticum is
present in the soil, but also in the gut of humans
and some animals as normal flora.
Diagenetic changes
Janaway [41] considered that soil was less important in the initial stages of decomposition, arguing
that the body will create its own environment which
in turn will modify the effects of the surrounding
soil. Studies in soil microbiology have shown that
24

this is not likely. Microorganisms are introduced

into farming soils so that certain growth characteristics of the soil may be improved. These microorganisms have been genetically manipulated to
give required characteristics. In almost all cases,
these introduced microorganisms die out; they do
so as a result of competition with the normal soil
flora, although some success has been achieved by
genetically manipulating soil isolates and re-introducing them into the same environment [78].
If a substrate (i.e., a corpse) which contains
microflora is added to soil, its own microflora will
be destroyed by competitive interactions with the
normal soil flora. This may take only a few months
or years, but the microorganisms present within the
corpse at death may induce diagenetic changes
before the indigenous population is destroyed.
Overall, the initial taphonomic changes induced in
a corpse may be the result of autolytic mechanisms
and inherent microbes. The most significant diagenetic changes seen in a corpse are more likely to be
due to the action of soil macro- and microflora.
A particular type of diagenetic change is that
seen in microscopical focal destruction (MFD).
These are localized 'tunnels' thought to be produced in bone tissue by the action of microorganisms (for a review, see Bell [79]). The dimensions of
the MFD are identical with the dimensions of
microorganisms; the microbial metabolites cannot
diffuse far into the dense bone tissue. In some types
of MFD, mineral redeposition along the interior
walls of the 'tunnels' has been noted. It is thought
that this is due to the death of the microorganisms:
when a microorganism dies, the pH of the surrounding medium rises due to the release of ammonia and other microbial metabolites. This rise in pH
will promote the re-precipitation of dissolved
hydroxyapatite.
An unexpected alteration of bone protein by the
bacterium Pseudomonasfluorescens has been shown
[77]. Decomposition studies using various strains of
this bacterium, both singly and in concert, has
shown that the bacterium has the facility to change
the rate of racemization of the aspartic acid
residues in the insoluble collagen of bone.
Post-excavation
Microbial communities are affected by environmental disturbances. The inherent microbial population
within archaeological bone probably receives its
greatest disturbance on excavation. The problems
created by the sudden alteration of redox potential
will be exacerbated by storage in a warm 'finds
tent'. The microbial loading within the bone will
already be significant; alteration of temperature
alone will increase microbial growth rates since,
generally, enzyme reaction rates are doubled for
Studies in Conservation40 (1995) 19-30

Microbialtaphonomyof archaeologicalbone
every 10?C rise in temperature[80]. The concomitantincreasein oxygen concentrationwill promote rapid aerobic decomposition of organic
components.Fast-growingfungi, previouslyheld in
check by burial conditions,tend to flourishat the
expense of other organisms.With the presence of
metabolitesfrom fungal action, the rate of decomposition of the bone will, to a certain extent, be
controlledby the interactionof the inherentmicrobial species (see 'Microbial interactions' below)
until physicalconditions(e.g. storage at controlled
relativehumidity)performthis function.
Chemical hydrolysis
If the mineral phase of bone retains its integrity,
then collagen can only be lost following chemical
alteration.A number of chemical transformations
will occur and will continueafter death, such as the
process of non-enzymiccrosslinking,but the most
significantchange will be hydrolysisof the peptide
bond.
A simple, conceptualmodel of collagen peptide
bone hydrolysishas been suggested.This model still
requires much work before it can be usefully
appliedto mineralizedcollagen,but it fits well with
many observedphenomena[81]. The rate of collagen loss from the bone will dependupon the rate of
peptide bond hydrolysis, the rate of diffusion of
these small fragmentsout of the bone and the rate
of post-mortemcrosslinking.
Crosslinkswill form as a result of the continued
non-enzymic process. 'Vegetable tannates' in the
burial environmentwill bond to the collagen [82].
The degree of bonding by these tannates will
depend upon the rate of diffusion of these chemicals into the bone. This diffusionwill be enhanced
following demineralizationof the bone. It is likely
that the presenceof these compoundsin the bone
will physically inhibit the action of collagenolytic
enzymes.
Limitations of microbial decomposition

matrix will fall. The fall in temperaturewill slow

microbialactivity.This slowingwill have a positive
effect on the fall in temperature,until the temperature of the decomposingmaterial reaches that of
the surroundingsoil.
Alkaline and acid soils
Alkaline soils will buffer the microbial acids and
inhibit the dissolution of the bone mineral.
Conversely,acidic soils will tend to promote bone
decomposition,although this is only a generalization. The rate of hydrolysis of peptide bonds is
increasedin both acid and alkalineenvironments.
Microbial interactions
Many bacteria and fungi generate by-productsof
metabolism that are antagonisticto other microorganisms(i.e., antibiotics). These chemicalshave
a suppressanteffect both on membersof the same
genusand on completelydifferentgenera[83, 84].
Generally,competition between microorganisms
tends to prolong the life of the substrate. The
greaterthe varietyof microorganismswithin a substrate, the longer it is likely to survive,since some
microbialenergymust be directedtowardscontrolling other microorganismsrather than substrate
digestionalone.
Non-microbial inhibitorysubstances
Burial environmentsmay inhibit the growth of
microorganismscapable of degrading bone. The
presenceof copper(I)-but probablynot iron(II)ions as well as other inhibitory substances (e.g.
vegetable tannates) may promote the survival of
both the soft and the hardtissues[85].
Extremes of temperature
Extremesof temperaturecan promote preservation
of both the soft and the hard tissues: they are
known to survivein desertsand permafrostregions
[37]. Desiccationwill reducemicrobialactivity and
hence the only mechanismfor collagendegradation
is chemical hydrolysis, the rate of which would
dependupon temperature.

Once microbialdecompositionis stopped, the only

degradation process occurring is chemical. The
causes of the reductionin microbialactivitymay be
multifactorial.If any one or a combinationof these Enclosed environments
conditions is met, then microbial decomposition Enclosed environmentswhere there is little or no
may slow or even stop, but chemicalhydrolysiswill leachingwill promotethe preservationof bone. An
continue.
equilibriumwill be reached between the microbial
products of metabolism and the bone mineral
Burial temperature
bufferingcapacity. The presence of the microbial
As the rate of decomposition of the soft tissue metabolites will inhibit further microbial growth,
slows, due to its removalby aerobicand anaerobic since they are toxic to the microorganisms,and
decomposition,the temperatureof the decomposing thus the microbialpopulationswill slowly decline.
Studiesin Conservation
40 (1995) 19-30

25

A.M. Child

Conclusion
The various pathways for the microbial taphonomy
of archaeological (and historical) bone have been
defined here and, even though an exhaustive list has
not been given, it can be seen that a bewildering
range of reactions is possible. These include demineralization of the bone, damage due to the action
of microbial enzymes (both collagenases and proteases), the microbial production of MFD (microscopical focal destruction) and the alteration of the
proteins following decomposition.
Acknowledgements
The author would like to thank Professor R.D.
Gillard, Dr Matthew Collins, Dave Watkinson,
Susan Hardman and Naomi Earl-Turner for their
helpful comments during the preparation of this
manuscript. Thanks are also due to Dr J. Morgan
and the staff of the Cardiff Royal Infirmary,
Bacteriology Department, for their kind permission
to use the laboratory facilities for the microbial isolation work. The project was funded by the Science
& Engineering Research Council (Science-Based
Archaeology Committee).
References
1 BUNN,M. K., 'Saran as a treatment for archaeological bone', undergraduate dissertation,
Cardiff (1987).
2 HUECK,H. J., 'The biodeterioration of materials-an appraisal' in The Biodeterioration of
Materials, ed. A. H. WALTERSand J. J.
ELPHICK,
Elsevier, London (1968) 6-12.
3 EFREMOV,
I. A., 'Taphonomy, a new branch of
paleontology', Pan-American Geologist 74
(1940) 81-93.
4 TRIFFITT,J. T., 'The organic matrix of bone' in
Fundamental and Clinical Bone Physiology,
ed. M. R. URIST, Lippincott, Philadelphia/
Toronto (1980).
5 GREEN,M., ISAAC,D. H., and JENKINS,G. M.,
'The structure of bone mineral and its spatial
relationship with collagen' in American
Society of Mechanical Engineers, Winter
Meeting, Boston 1987 (1987) 59-60.
6 MISRA,D. N., 'Surface chemistry of bone and
tooth mineral' in Methods of Calcified Tissue
Preparation, ed. G. R. DICKSON,Elsevier,
Amsterdam/New York/Oxford (1984).
7 OAKLEY,K. P., and HOSKINS,C. R., 'New evidence on the antiquity of Piltdown Man',
Nature 165 (1950) 379-382.
26

8 VEIS,A., and SABSAY,B., 'The collagen of mineralised matrices' in Bone and Mineral
Research 5, Elsevier Science Publishers,
Amsterdam (1987) 1-64.
9 MILLER,E. J., and GAY, S., 'Collagen: an
overview', Methods in Enzymology 82 (1982)
33-64.
10 AITKEN,A., GEISOW,M. J., FINDLAY,J. B. C.,
HOLMES,C., and YARWOOD,A., 'Peptide
preparation and characterisation' in Protein
Sequencing: A Practical Approach, IRL Press,
Oxford (1989) 43-68.
11 LEES, S., 'Some characteristics of mineralised
collagen' in Calcified Tissue: Topics in
Molecular and Structural Biology, ed. D. W.
HUKINS, Macmillan Press, London (1989)
153-174.
12 GLIMCHER,
M. J., and KATZ,E. P., 'The organisation of collagen in bone; the role of noncovalent bonds in the relative insolubility of
bone collagen', Ultrastructural Research 12
(1965) 705-729.
13 INGRAM,R. T., CLARKE,B. L., FISCHER,L.W.,
and FITZPATRICK,
L. A., 'Distribution of
noncollagenous proteins in the matrix of
adult human bone. Evidence of anatomic and
functional heterogeneity', Journal of Bone and
Mineral Research 8(9) (1993) 1019-1029.
14 POTTS,R., and SHIPMAN,P., 'Cutmarks made
by stone tools on bones from Olduvai Gorge,
Tanzania', Nature 291 (1981) 577-580.
15 GRUPE, G., and GARLAND, A. N., (eds),
Histology of Ancient Human Bone. Methods
and Diagnosis
of
the
(proceedings
'Palaeopathology Workshop', Gottingen, 3-5
October 1990), Springer-Verlag, Berlin (1993).
16 HASSAN,A. A., and HARE, P. E., 'Amino acid
analysis in radiocarbon dating of bone collagen' in Archaeological ChemistryII, American
Chemical Society (1978) 108-116.
17 HEDGES,R. E. M., and LAW,I. A., 'The radiocarbon dating of bone', Applied Geochemistry
4 (1989) 249-253.
18 LONGIN,R., 'New method of collagen extraction for radiocarbon dating', Nature 230
(1971) 241-242.
19 BADA,J. L., 'Amino acid racemization dating of
fossil bones', Annual Reviews of Earth and
Planetary Science 13 (1985) 241-268.
20 BELLOUMINI,G., and BADA, J. L., 'Isoleucine

epimerisation ages of the dwarf elephants of

Sicily', Geology 13 (1985) 451-452.
21 DENNISON, K. J., and HOUGHTON, P., 'Amino

acids in archaeological bone (2)', Journal of

Archaeological Science 13 (1986) 393-401.
22 ELSTER, H.,

GIL-AV,

E.,

and

WEINER, S.,

'Amino acid racemization of fossil bone',

Studies in Conservation40 (1995) 19-30

Microbial taphonomy of archaeological bone

23

24

25

26

27

28

29

30

31

32

33
34

35

Journal of Archaeological Science 18 (1991)

605-617.
AMBROSE,S. H., 'Effects of diet, climate and
physiology on nitrogen isotope abundances in
terrestrial foodwebs', Journal of Archaeological Science 18 (1991) 293-317.
DENIRO,M. J., and EPSTEIN, S., 'Influence of
diet on the distribution of carbon isotopes in
animals', Geochimica et Cosmochimica Acta
45 (1981) 341-351.
AMBROSE,S. H., and NORR, L., 'Experimental
evidence for the relationship of the carbon
isotope ratios of whole diet and dietary protein to those of bone collagen and carbonate'
in Prehistoric Human Bone. Archaeology at
the Molecular Level, Springer-Verlag, Berlin
(1993) 1-38.
HARE,P. E., 'Organic geochemistry of bone and
its relation to the survival of bone in the natural environment' in Fossils in the Making,
University of Chicago Press (1980) 208-219.
HEDGES,R. E. M., and WALLACE,C. J. A.,
'The survival of protein in bone' in
Biogeochemistry of the Amino Acids, Wiley,
New York (1980) 35-40.
THUESON,I., and ENGBERG,J., 'Recovery and
analysis of human genetic material from
mummified tissue and bone', Journal of
Archaeological Science 17 (1990) 679-689.
PAABO,S., HIGUCHI,R. G., and WILSON,A. D.,
'Ancient DNA and the polymerase chain
reaction: the emerging field of molecular
archaeology', Journal of Biological Chemistry
264 (1989) 1-4.
SILLEN,A., SEALY,J. C., and VAN DER MERWE,
N. J., 'Chemistry and palaeodietary research:
no more easy answers', American Antiquity 54
(1989) 504-512.
VON ENDT, D. W., and ORTNER, D. J.,
'Experimental effects of bone size and temperature on bone diagenesis', Journal of
Archaeological Science 11 (1984) 247-253.
MASTERS,P. M., 'Preferential preservation of
noncollagenous protein during bone diagenesis: implications for chronometric and stable
Geochimica et
measurements',
isotope
CosmochimicaActa 51 (1987) 3209-3214.
WYCKOFF,R. W. G., The Biochemistry of
Animal Fossils, Scientechnica Ltd, Bristol
(1972).
ASCENZI,A., BRUNORI,M., CITRO, G., and
ZITO,R., 'Immunological detection of hemoglobin in bones of ancient Roman times and
Eneolithic ages', Proceedings of the National
Academy of Sciences USA 82 (1985)
7170-7172.
CATTANEO,
K., PHILLIPS,P.,
C., GELSTHORPE,

Studies in Conservation40 (1995) 19-30

and SOKOL,R. J., 'Blood in ancient human

bones', Nature 347 (1990) 339.
36 HUQ, N. L., RAMBAUD,S. M., TEH, L. C.,
DAVIES,A. D., MCCULLOCH,
B., TROTTER,
M. M., and CHAPMAN,G. E., 'Immunochemical detection and characterisation of
osteocalcin from Moa bone', Biochemical and
Biophysical Research Communications 129
(1985) 714-720.
37 LOWENSTEIN,
J. M., 'Immunological reactions
from fossil material', Philosophical Transactions of the Royal Society of London B 292
(1981) 143-149.
38 SMITH, P. R., and WILSON, M. T., 'Detection

of

haemoglobin in human skeletal remains by

ELISA', Journal of Archaeological Science 17
(1990) 255-268.
39 TUROSS, N., 'Albumin preservation in the
Taima-taima mastodon skeleton', Applied
Geochemistry4 (1989) 255-259.
40 CHILD, A. M., and POLLARD, A. M., 'A review

of the applications of immunochemistry to

archaeological bone', Journal of Archaeological Science 19 (1992) 39-47.
41 JANAWAY,
R. C., 'Dust to dust: the preservation
of textile materials in metal artefact corrosion
products with reference to inhumation
graves', Science and Archaeology 27 (1985)
29-34.
42 O'CONNOR, T. P., 'On the structure, chemistry

43

44
45
46

47

and decay of bone, antler and ivory' in

Archaeological Bone, Antler and Ivory,
Occasional Paper No. 5, United Kingdom
Institute for Conservation, London (1987)
6-8.
R. C. A., 'Variation in the chemiROTTLANDER,
cal composition of bone as an indicator of
climatic change', Journal of Archaeological
Science 3 (1976) 83-88.
WHITE,E. M., and HANNUS,L. A., 'Chemical
weathering of bone in archaeological soils',
American Antiquity 48 (1983) 316-322.
LYNCH, J. M., and POOLE,N. J., Microbial
Ecology, A Conceptual Approach, Blackwell,
Oxford (1979) 72-73.
HENDERSON,
J., 'Factors determining the state
of preservation of human remains' in Death,
Decay and Reconstruction, Manchester University Press (1987) 43-54.
PAUL, E. A., and CLARKE,F. E., Soil Microbiology and Biochemistry, Academic Press,
San Diego (1989) 11-31.

48 GRUPE, G., and PIEPENBRINK, H., 'Trace ele-

ment contamination in excavated bones by

microorganisms' in Trace Elements in
Environmental History,
Springer-Verlag,
Berlin (1988) 103-112.
27

A.M. Child
49 GRUPE, G., and PIEPENBRINK,
H., 'Impact of
microbial activity on trace element concentrain
tions
excavated
bones',
Applied
Geochemistry4 (1989) 293-298.
50 GOTTSCHALK, G.,
Bacterial Metabolism,
Springer-Verlag, New York (1986).
51 MARGOLIS,H. C., and MORENO, E. C.,
'Kinetics of hydroxyapatite dissolution in
acetic, lactic and phosphoric acid solutions',
Calcified Tissue International 50 (1992)
137-143.
52 BRIGGS,D. E. G., and KEAR,A. J., 'Decay and
preservation of polychaetes: taphonomy
in
soft-bodied
thresholds
organisms',
Palaeobiology 19(1) (1993) 107-135.
53 BARMAN, T. E., 'Collagenases' in Enzyme
Handbook, 2nd edn, Vol. II, Springer-Verlag,
Berlin/Heidelberg (1985) 636.
54 WEINGARTEN,
H., and FEDER,J., 'Cleavage site
specificity of vertebrate collagenases', Biochemical and Biophysical Communications139
(1986) 1184-1187.
C. A.,
55 BERRY, L., and SHUTTLEWORTH,
'Bacterial collagenase and collagen identification', Connective Tissue Research 17 (1988)
153-158.
56 BOND, M. D., and VAN WART, H. E.,
'Purification and separation of individual collagenases of Clostridium histolyticum using
red dye ligand chromatography', Biochemistry
23 (1984) 3077-3085.
57 CHAKRABORTY,
R., and CHANDRA, A. L.,
'Purification and characterisation of a streptomycete collagenase', Journal of Applied
Bacteriology 61 (1986) 331-337.
58 ENDO, A., MURAKAWA,
S., SHIMUZU,H., and
SHIRAISHI,Y., 'Purification and properties of

59

60

61

62

28

collagenases from a Streptomyces species',

Journal of Biochemistry 102 (1987) 163-170.
MAKINEN, K. K., and MAKINEN, P. L.,
'Purification and properties of an extracellular collagenolytic protease produced by the
human oral bacterium Bacillus cereus (Strain
Soc 67)', Journal of Biological Chemistry 262
(1987) 12488-12495.
BOND, M. D., and VAN WART, H. E.,
'Characterisation of the individual collagenases of Clostridiumhistolyticum', Biochemistry
23 (1984) 3085-3091.
ANGELTON,E. L., and VAN WART, H. E.,
'Preparation and reconstitution with divalent
metal ions of Class I and Class II Clostridium
histolyticum apocollagenases', Biochemistry 27
(1988) 7406-7412.
VAN WART,H. E., and BIRKEDAL-HANSEN,
H.,
'The cysteine switch: a principle of regulation
of metalloproteinase activity with potential

63

64

65

66

67

68

69

70

71

72

applicability to the entire matrix metalloprotease gene family', Proceedings of the

National Academy of Science USA 87 (1990)
5578-5582.
KRANE,S., 'Degradation of collagen in connective tissue diseases: rheumatoid arthritis' in
Dynamics of Connective Tissue Macromolecules, North Holland Publishing Co.
(1974) 309-326.
TSAI, I. H., Lu, P.-J., and CHUANG,J.-I., 'The
midgut of shrimps Penaeus monodon, Penaeus
japonicus and Penaeus penicillatus', Biochimica et Biophysica Acta 1080 (1991) 59-67.
WELGUS,H. G., GRANT,G. A., JEFFERY,J. J.,
and EISEN,A. Z., 'Substrate specificity of the
collagenolytic serine protease from Uca pugilator. Studies with collagenous substrates',
Biochemistry 21 (1982) 5183-5189.
KATZ, J., and BIRKEDAL-HANSEN,H.,
'Degradation of reconstituted collagen fibrils
at acidic pH', Journal of Dental Research 65
(1986) 183.
MILLER,E. J., and RHODES,R. K., 'Preparation
and characterisation of the different collagen
types', Methods in Enzymology 82 (1982)
33-64.
CHILD,A. M., 'Towards an understanding of
the microbial decomposition of archaeological bone in the burial environment' in proceedings of the Oxford Bone Diagenesis
Meeting (July 1993) for Journal of
Archaeological Science (in press).
E. A., 'Factors affecting soil forFITZPATRICK,
in
mation'
Their Formation,
Soils,
Classification and Distribution, Longman,
London (1983) 27.
CATO, E. P., GEORGE,W. L., and FINEGOLD,
S. M., 'Clostridium' in Bergey's Manual of
Williams
&
Bacteriology,
Systematic
Williams, Baltimore (1986) 1141-1200.
CHILD,A. M., GILLARD,R. D., and POLLARD,
A. M., 'Microbially-induced racemization:
isolation of the microorganisms and demonstration of their enzymes', Journal of
Archaeological Science 20 (1993) 159-164.
GARLAND,A. N., 'Microscopical analysis of fossil bone', Applied Geochemistry 4 (1987)
109-126.

73 TRUMP, B. F., BEREZSKY,I. K., and OSORNIO-

VARGAS,A. R., 'Cell death and the disease

process. The role of calcium' in Cell Death in
Biology and Pathology, Chapman Hall,
London (1981).
74 HAGIHARA, Y., KAMINISHI, I. K., CHO, T.,
TANAKA, M., and KAITA, H., 'Degradation

of the human dentine collagen by an enzyme

produced by the yeast Candida albicans',
Studies in Conservation40 (1995) 19-30

Microbial taphonomy of archaeological bone

75

76

77

78

79

80
81

Archives of Oral Biology 33(8) (1988)

617-619.
WALDREN,T. E., 'The relative survival of the
human skeleton: implications for palaeopathology' in Death, Decay and Reconstruction, Manchester University Press (1987)
55-64.
BODDINGTON,
A., 'Chaos, disturbance and decay
in an Anglo-Saxon cemetery' in Death, Decay
and Reconstruction, Manchester University
Press (1987) 27-42.
CHILD, A. M., 'Microbially-induced racemization of aspartic acid: decomposition studies',
in preparation for Journal of Archaeological
Science.
VAN ELSAS, J. D., TREVORS,J. T., VAN
OVERBEEK,L. S., and STARODUB,M. E.,
'Survival of Pseudomonas fluorescens containing plasmids RP4 or pRK2501 and plasmid
stability after introduction into two soils of
different textures', Canadian Journal of
Microbiology 35 (1989) 951-959.
BELL,L. S., 'Palaeopathology and diagenesis; an
SEM evaluation of structural changes using
backscattered electron imaging', Journal of
Archaeological Science 17 (1990) 85-102.
CHAPLIN, M. F., and BUCKE, C., Enzyme
Technology, Cambridge University Press
(1990) 18-19.
COLLINS,M. J., RILEY,M., CHILD,A. M., and
TURNER-WALKER,
G., 'The potential importance of chemical degradation to the analysis
of ancient collagen' in proceedings of the
Oxford Bone Diagenesis Meeting (July 1993)
for Journal of Archaeological Science (in
press).

82 HARLAN, J. W., and FAIERHELLER,

S. H.,
'Chemistry of the cross-linking of collagen
during tanning', Advances in Experimental
Medicine and Biology 86A (1977) 425-440.
83 RAYNOR,A. D. M., and BODDY, L., Fungal
Decomposition of Wood. Its Biology and
Ecology, Wiley & Sons, Chichester (1990).
84 YOUNG, A. M., 'Microbial activity in waterlogged wood' in Preprints for the 30th
Anniversary Conference: Conservation Today,
United Kingdom Institute for Conservation,
London (1988) 123-127.
85 BROWN,N. L., LEE, T. 0., and SILVER,S.,
'Bacterial transport of and resistance to copper' in Metal Ions in Biological Systems 30 (in
press).

Author
ANGELA M. CHILD studied microbiology at the

University of London and worked for several years

within the National Health Service. She went on to
study archaeological conservation at the University
of Wales College of Cardiff, graduating in 1988.
Her special interest was the decomposition of
organic materials and she completed her PhD on
aspects of bone decomposition in 1992, followed by
a two-year SERC (SBAC) post-doctoral fellowship
which continued the investigation of the interaction
of microbial populations in the decomposition of
archaeological bone. She is currently working in the
field of ancient biomolecules, on the interaction of
osteocalcin with hydroxyapatite. Address: Fossil
Fuels & Environmental Geochemistry, NRG,
Drummond Building, University of Newcastle,
Newcastle upon Tyne NE1 7RU, UK.

Resum--La taphonomie est l'etude de tout changement qui intervientsur un animal ou sur -uneplante apres sa
mort. Les os sont les principaux materiaux d'origine animale d survivre dans l'environnementarcheologique.
C'est une source importante d'informations sur les relations, les regimes, les maladies et les dges des peuples
(et des animaux) des anciennes cultures. Pour trouver et interpreter ces informations, il est besoin de conservateurs et d'hommes de sciences archeologues qui connaissent les processus de changements taphonomiquesqui
peuvent advenir aux os. Ces changements sont nombreux: ce papier tente de definir quelques-unsde ces principaux changements, qui peuvent etre attribues a l'action variee des microorganismes.
Zusammenfassung-Taphonomie bezeichnet das Studium aller Veranderungenan Lebewesen und Pflanzen
nach dem Tod. Knochen sind das vorherrschendeMaterial lebenden Ursprunges, das sich im archaologischen
Rahmen erhalt. Sie sind die Quelle fir eine Vielzahl von Informationen uiber Beziehungen, Nahrung,
Krankheiten und Alter von Menschen und Tieren vergangenerKulturen. Damit diese Informationengesammelt
und interpretiertwerden k6nnen, miissen Konservatorenund archdologische Wissenschaftlerein Bewujftseinfur
taphonomische Veranderungen entwickeln, die sich in Knochenmaterial abspielen konnen. Die vorliegende
Arbeit versucht, wichtigste in Knochen feststellbare Veranderungenzu definieren, welche den zahlreichen
Aktivitdten von Mikroorganismenzuzuschreibensind.
Resumen-Tafonomia es el estudio de todos los cambios efectuados en las sustdncias animales y vegetales
despues de la muerte de las mismas. De los restos de origen animal, el hueso es el principal material perStudies in Conservation40 (1995) 19-30

29

A.M. Child
durable dentro del dmbito arqueol6gico. El hueso es una abundantefuente de informacion acerca de las relaciones sociales, dietas, enfermedades y edades de gentes (y animales) de culturas pasadas. Para extraer e
interpretaresta informacion, es necesario que los conservadoresy cientificos arqueol6gicos esten conscientes de
los procesos de cambio tafonomicos que pueden ocurir en el hueso. Estos procesos de cambio son numerosisimos. Este trabajopropone la definicion de algunos de los principales cambios observados en hueso y atribuibles
a las acciones varias de los microorganismos.

30

Studies in Conservation40 (1995) 19-30