Booth 2015 Archaeometry

7242019 Booth 2015 Archaeometry

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A N I N V E S T I G A T I O N I N T O T H E R E L A T I O N S H I P B E T W E E N

F U N E R A R Y T R E A T M E N T A N D B A C T E R I A L B I O E R O S I O N I N

E U R O P E A N A R C H A E O L O G I C A L H U M A N B O N E

T J BOOTHdagger

Department of Earth Sciences Natural History Museum Cromwell Road London SW7 5BD UK

A central problem in funerary archaeology is interpreting how the corpse was manipulated in

the immediate post mortem period The extent of bacterial bioerosion to the internal bone

microstructure has been proposed as a means to infer the early post mortem history of a

corpse as it has been suggested that this form of bone diagenesis is produced by an organ-

ismrsquos putrefactive gut bacteria Under this model different forms of funerary treatment would

be expected to leave characteristic signatures of bioerosion in archaeological bone Here wetested the extent to which bacterial bioerosion of ancient human bones re 1047298 ected funerary

treatment through histological analysis of 301 archaeological human bone thin sections from

25 European archaeological sites We found that bioerosion was signi 1047297cantly in 1047298 uenced by

whether a bone originated from a neonatal individual or an anoxic context When these

remains were excluded bioerosion was controlled by archaeological phase in a manner

consistent with known early post mortem treatment and forensic models of bodily decomposi-

tion These 1047297ndings suggest that microscopic analyses of bone have useful applications in

reconstructions of funerary processes and provide some insight into factors that may control

the persistence of organic biomolecules and fossilization

KEYWORDS BONE DIAGENESIS TAPHONOMY FUNERARY TREATMENT NORTH-WESTEUROPE THIN-SECTION LIGHT MICROSCOPY

INTRODUCTION

After the death of a vertebrate organism the skeleton is subject to a variety of physico-chemical

changes that lead to its destruction or fossilization (Hedges 2002 Trueman and Martill 2002

Nielsen-Marsh et al 2007 Smith et al 2007 Lee-Thorp and Sealy 2008) Microbial bioerosion

is the most common diagenetic change observed within archaeological bone (Hackett 1981

Nielsen-Marsh and Hedges 2000 Hedges 2002 Turner-Walker et al 2002 Jans et al 2004

Nielsen-Marsh et al 2007) Hackett (1981 250) de1047297ned four categories of micro-foci of destruc-tion (MFD) that constitute bioerosion Inoculation experiments established that Wedl tunnelling

is caused by saprophytic fungi (Marchiafava et al 1974 Fernaacutendez-Jalvo et al 2010) and similar

experiments combined with microscopic analyses of archaeological bone determined that all

three forms of non-Wedl MFD are produced by bacteria (Yoshino et al 1991 Balzer et al

1997 Grupe and Turban-Just 1998 Jackes et al 2001 Turner-Walker and Syversen 2002

Dixon et al 2008) Bacterial bioerosion is the predominant form of microbial attack observed

within archaeological bone (Nielsen-Marsh and Hedges 2000 Turner-Walker et al 2002

Hedges 2002 Jans et al 2004 Nielsen-Marsh et al 2007)

Received 28 August 2014 accepted 2 February 2015

daggerCorresponding author email tboothnhmacuk

Archaeometry bullbull bullbull (2015) bullbullndashbullbull doi 101111arcm12190

copy 2015 The Authors

Archaeometry published by John Wiley amp Sons Ltd on behalf of University of Oxford

This is an open access article under the terms of the Creative Commons Attribution License which permits use distribution and

reproduction in any medium provided the original work is properly cited

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Early theories regarding the aetiology of bacterial bone bioerosion were concerned with soil

micro-organisms (Marchiafava et al 1974 Hackett 1981 Piepenbrink 1986 1989 Hanson

and Buikstra 1987 Yoshino et al 1991 Grupe and Dreses-Werringloer 1993) Successive

studies have suggested that non-Wedl MFD are produced by a dead organism rsquos gut bacteria (Child

1995a Bell et al 1996 Jans et al 2004 Guarino et al 2006 Nielsen-Marsh et al 2007 Hollundet al 2012 White and Booth 2014) The post mortem deterioration of an organism rsquos immune sys-

tem and mucosal membranes facilitate the transmigration of gut bacteria within the 1047297rst few days

after death (Polson et al 1985 Janaway 1987 Child 1995ab Gill-King 1997) These micro-

organisms are primarily responsible for bodily putrefaction (Polson et al 1985 Janaway 1987

Child 1995ab) An endogenous origin of osteolytic microbiota suggests that bacterial bone

bioerosion should re1047298ect the extent to which the skeleton was exposed to putrefaction

This hypothesis presents a potentially useful prospect for the study of funerary archaeology

Different post mortem treatments should expose the skeleton to divergent levels of putrefaction

and leave characteristic signatures of bacterial bioerosion within the bone microstructure (Bell

et al 1996 Jans et al 2004 Parker Pearson et al 2005 Hollund et al 2012 White and Booth2014) Certain studies have already detected logical relationships between bacterial bone

bioerosion and early taphonomic events (Bell et al 1996 Jans et al 2004 Nielsen-Marsh

et al 2007 White and Booth 2014)

Measures of bacterial bioerosion in archaeological human bone combined with models of de-

composition could help to distinguish between discrete funerary rites that leave similar archaeo-

logical records or help to identify hidden complexities in funerary treatment (Bell et al 1996

Jans et al 2004 Parker Pearson et al 2005 Nielsen-Marsh et al 2007 Smith et al 2007

Turner-Walker and Jans 2008 Hollund et al 2012) Furthermore microbial exploitation is a pri-

mary mechanism of bone collagen loss and has been variably linked with successful extraction of

organic biomolecules including DNA (Hagelberg et al 1991 Grupe 1995 Colson et al 1997Cipollaro et al 1998 Geigl 2002 Goumltherstroumlm et al 2002 Haynes et al 2002 Rollo et al

2002 Collins et al 2009 Ottoni et al 2009 Deviegravese et al 2010) Establishing the factors that

affect bone bioerosion may help to account for variation in organic biomolecular yield amongst

archaeological bones and contribute to predictive models of preservation

There have been no systematic investigations into whether speci1047297c funerary treatments pro-

duce predictable signatures of bacterial bioerosion within archaeological human remains It has

yet to be determined whether the effects of anthropogenic processes can be distinguished from

changes elicited by the burial environment Seasonality is often cited as the most signi1047297cant fac-

tor that affects the nature of bodily decomposition (Rodriguez and Bass 1983 1985 Mann et al

1990 Manhein 1997 Campobasso et al 2001 Wilson et al 2007 Zhou and Bayard 2011Meyer et al 2013) Anoxic burial environments can arrest bodily putrefaction and have been

consistently observed to discourage bacterial bioerosion of bone microstructures (Polson et al

1985 Cotton et al 1987 Mant 1987 Janaway 1996 Turner and Wiltshire 1999 Fielder and

Graw 2003 Wilson et al 2007 Turner-Walker and Jans 2008 OrsquoConnor et al 2011 Hollund

et al 2012) The endogenous model of bacterial bioerosion and the in1047298uence of early taphonomic

events have been questioned more recently (Turner-Walker 2008 2012 Fernaacutendez-Jalvo et al

2010) The strength of the relationship between early post mortem processes and bacterial bone

bioerosion is questionable and needs to be explored further to determine how it might be used in

reconstructions of funerary activities in the past

In order to establish the relationship between bacterial bone bioerosion and funerary treat-ment and to assess whether microscopic analysis of archaeological human remains could aid

reconstructions of funerary treatments practised by past populations we conducted

2 T J Booth


Archaeometry published by John Wiley amp Sons Ltd on behalf of University of Oxford Archaeometry bullbull bullbull (2015) bullbullndashbullbull



microscopic analysis of archaeological human bones recovered from European later prehis-

toric (4000 BC ndash AD 43) and British historical (AD 43 ndash present day) sites These two assem-

blages were used as proxies for funerary treatment The historical British archaeological

record represents a period of time in which the majority of people were consistently buried

soon after death Burial protects the body from invertebrates that promote rapidskeletonization such as Diptera ensuring that bones experience the maximal levels of putre-

factive activity (Rodriguez and Bass 1983 1985 Mann et al 1990 Campobasso et al 2001

Breitmeier et al 2005 Simmons et al 2010 Zhou and Bayard 2011 White and Booth 2014)

Human remains from European later prehistoric sites are often recovered in variable states of

articulation and numerous strands of archaeological evidence have been used to infer that

these individuals were subject to highly variable practices (Parker Pearson 1999 Parker

Pearson et al 2005 Redfern 2008 Darvill 2010) Any funerary practice that did not involve

immediate burial after death would be expected to variably reduce the levels of putrefaction

experienced by a skeleton Comparison of bacterial bioerosion within human remains from

historical and later prehistoric sites provided a rudimentary but effective scrutiny of the rela-tionship between funerary treatment and bacterial bioerosion by testing a simple dichotomy of

variable and consistent treatment Therefore we devised three predictions based on the as-

sumption that bone bioerosion is related to funerary treatment as de1047297ned by models of bodily

decomposition

(1) Archaeological phase (later prehistoric versus historical) has a signi1047297cant independent

in1047298uence on bacterial bone bioerosion

(2) Historical bone samples demonstrate consistently high levels of bacterial bioerosion

(3) Later prehistoric bone samples demonstrate variable levels of bacterial bioerosion

MATERIALS AND METHODS

Materials

The histological preservation of archaeological human bone thin sections from 301 individuals

retrieved from 25 European sites was assessed using conventional light microscopy (Table S1

and Fig 1) A proportion of these samples (46) were not produced speci1047297cally for our study

but constituted the University of Shef 1047297eldrsquos human bone thin-section collection The remaining

thin sections were produced from the University of Shef 1047297eldrsquos archaeological collections as well

as from a series of samples accessed from various European institutions For historical periods

we focused on simply obtaining bone samples from dispersed sites while sampling of prehistoricremains also attempted to capture variation in post mortem treatment as determined by state of

skeletal articulation and evidence for post mortem manipulation (eg cut marks)

Bacterial bioerosion can vary between skeletal elements (Hanson and Buikstra 1987 Bell

et al 1996 Jans et al 2004) Long bones are usually chosen for histological analysis due to their

cortical bone content robusticity and survival rate We preferentially targeted femora to maxi-

mize comparability with previous studies (Nielsen-Marsh and Hedges 2000 Jans et al 2004

Hollund et al 2012) When sampling from disarticulated assemblages femora from the same

side of the body were taken consistently to ensure that there was no replication of individuals

The volume of human bone recovered from later prehistoric contexts was sometimes too low

to acquire a good sample size from femora alone and samples had to be taken from alternativelong bones Some of these bones originated from disarticulated assemblages but in each case

the taphonomic and contextual evidence was scrutinized to con1047297rm that every sample

Funerary treatment and bacterial bioerosion in human bone 3





represented a separate individual Some of the University of Shef 1047297eldrsquos thin sections had been

taken from non-femoral skeletal elements All thin sections included in the current study came

from long bones and the majority (97) originated from femora It was unlikely that the non-

femoral samples would have affected overall 1047297ndings although the effect of different skeletal el-ements was monitored

Preparation and assessment of thin sections

Samples around 1 cm times 1 cm were cut from the mid-section of each long bone diaphysis using a

Foredom K1070 rotary saw Transverse thin sections 50ndash120μm thick were produced from

these samples using a Leica 1600 diamond-saw microtome Particularly friable samples were em-

bedded using Araldite 2020 (Huntsman Advanced Material) or LR White Acrylic resin (Agar

Scienti1047297c) Each undecalci1047297ed and unstained thin section was mounted on to a glass slide using

Entellan (Merck Chemicals) or Euparal (Alpha Chemika) All sections were analysed under nor-mal and polarized light using transmitted light binocular microscopes 1047297tted with polarizing 1047297l-

ters at 25times 40times and 100times magni1047297cation Bacterial bioerosion was assessed using the standard

Oxford Histological Index (OHI) (Hedges et al 1995 Millard 2001) Additionally the occur-

rence of bacterial attack was recorded on a presenceabsence basis to document variation in its

appearance as well as its extent All statistical tests were carried out using IBM SPSS

Other potentially in 1047298 uential variables

The speci1047297city of the predictions set out above ensured that they would be refuted if bacterial

bioerosion was not signi1047297

cantly in1047298

uenced by funerary treatment However we attempted tocontrol for other potential factors to mitigate the possibility of false results The precise effects

of these variables on bacterial bioerosion are ill-de1047297ned and their in1047298uence had to be gauged

Figure 1 The distribution of the later prehistoric (white triangles) and historical (grey circles) archaeological sites in-

cluded in the current study

4 T J Booth





on a post hoc basis There is con1047298icting evidence over whether soil composition affects bone

bioerosion either through the constitution of soil bacteria or interference with bodily decompo-

sition (Hanson and Buikstra 1987 Nielsen-Marsh et al 2007 Turner-Walker and Jans 2008

Turner-Walker 2008 2012) Species of collagenase-producing bacteria capable of exploiting

bone protein are found commonly within most types of soil (Vranyacute et al 1988) However it isstill unclear whether these bacteria produce characteristic MFD (Balzer et al 1997) A large pro-

portion of these soil bacteria may be unable to produce collagenase in the low temperatures of the

burial environment (Child 1995b)

Burial sediments could be split into one of four broad categories sand silt clay gravel and

open (no sediment) These categories crudely represented collective features of environments that

affect early bodily decomposition and the abundance and composition of resident bacteria such

as matrix potential composition and density (Janaway 1996 Dent et al 2004 Carter et al

2008 2010 Jagger and Rogers 2009) The survival of archaeological bones at these sites

suggested that soil pH varied little from slightly acidic to alkaline (Gordon and Buikstra

1981 Bethel and Carver 1987 Nielsen-Marsh and Hedges 2000 Nielsen-Marsh et al 2007Smith et al 2007)

Anaerobic conditions were recorded on a presenceabsence basis in cases where the burial sed-

iment was intrinsically anoxic where there was evidence that the sediment had been waterlogged

previously andor where there was extraordinary survival of organic material A proportion

(42) of the historical thin sections originated from disarticulated charnel bone recovered from

grave 1047297lls or formal charnel deposits It was likely that these remains represented individuals that

had been buried and disturbed post-skeletonization (Daniell 1997) However whether a bone

sample originated from a charnel deposit was recorded to ensure that this factor had no effect

on bacterial bioerosion

The progressive age-related increase in bone porosity attributable to cumulative secondaryosteon formation (remodelling) may render bones from older individuals more prone to bacte-

rial invasion (Turner-Walker et al 2002 Turner-Walker 2008) In addition the sterility of

mammalian intestinal tracts before birth (Mackie et al 1999 Sharon et al 2013 Jakobsson

et al 2013) has also been proposed as a factor in eliminating bioerosion of neonates

(Economou 2003 White and Booth 2014) All of the skeletons whose bones were used in

the present study had been assessed using modern standard osteological methods (Buikstra

and Ubelaker 1994 Bass 1995 Schwartz 1995 Cox and Mays 2000 Scheuer and Black

2000 Brickley and McKinley 2004)The inclusion of disarticulated long bones meant that

many specimens could only be assigned broad age-at-death estimates All samples were classi-

1047297ed into one of four categories of neonate (foetal or less than 1 month) child (between 1 monthand 10 years) juvenile (between 11 and 18 years) and adult (over 18 years) This methodology

suited the aims of our study as all posited age-related variations in bone bioerosion describe

differences between sub-adult and adult remains (Jans et al 2004 Turner-Walker 2008 White

and Booth 2014)

Some of the bone samples originated from the medieval cemetery uncovered at East

Smith1047297eld London (Grainger et al 2008) A proportion of these skeletons came from a ceme-

tery that had been founded in response to the AD 1348ndash50 Black Death epidemic Some of the

Black Death skeletons had been recovered in variable states of articulation from undisturbed

graves which indicated that there had been a delay between their death and burial Levels of

putrefaction experienced by these remains may have deviated from those encouraged by imme-diate burial Samples of bone from the Black Death cemetery were recorded to account for this

possibility






RESULTS

The Oxford histological index

The OHI categorizes underlying continuous data therefore a normal distribution was taken to

represent insigni1047297cant variation The highest proportion of samples (42) demonstrated OHI

scores of zero All of the non-zero OHI scores were replicated as negative numbers to create a

distribution around scores of 0 (Fig 2) There was a signi1047297cant difference between this distribu-

tion and an idealized normal model which suggested that there was notable variation in the OHI

score (n = 475 KolmogorovndashSmirnov Z = 2914 p = 0000)

OHI score variation amongst all but one sample was attributable to non-Wedl MFD The

anomalous specimen demonstrated no bacterial tunnelling but had lost a small proportion of

its microstructure to Wedl-type attack The small number and variety of non-femoral samples

meant that variation in bacterial bioerosion with skeletal element could not be tested statistically

However OHI scores were never observed to vary with skeletal element amongst our sample set

and there was no justi1047297cation for excluding them from the analysisThe OHI scores were placed in an ordinal regression model to determine which variables had

independently in1047298uenced variation in bacterial bioerosion Age at death anoxic environment

whether a bone originated from a Black Death cemetery and archaeological phase all had a sig-

ni1047297cant effect on the OHI score (Table 1) This ordering re1047298ected the descending in1047298uence of

each variable as de1047297ned by associated parameter estimates The OHI data are displayed as

jittered scatter graphs alongside conventional box-and-whisker plots to better visualize the dis-

tribution density of all cases The data were jittered in SPSS by adding a randomly generated

number between 0 and 09 to each OHI score Neonatal remains explained the majority of the

variation in OHI score with age at death (Fig S1) The transition between child and juvenile

ages also had a signi1047297cant in1047298uence on the OHI score but the parameter estimate for this out-come was smaller than those produced for the other variables Neonatal samples demonstrated

higher OHI scores than post-neonatal specimens The histological preservation of bone samples

Figure 2 The distribution of the OHI scores amongst the whole study sample when all non-zero values were replicated

as negative numbers with a superimposed normal curve

6 T J Booth





from anoxic environments was generally higher and more varied than within bone samples from

aerobic contexts (Fig S2) Samples of bone from Black Death graves demonstrated higher and

more variable OHI scores than those from nonndashBlack Death contexts (Fig S3) Bones from later

prehistoric contexts demonstrated higher and more variable OHI scores compared to historicalspecimens (Fig 3)

Table 1 Results of the ordinal logistic regression model applied to the OHI score signi 1047297cant p-values are

highlighted in bold

Variable Outcome

Number of

samples

Parameter

estimate

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)

Anoxia Absent 261 2065 0367 31732 lt00005

Present 40 0 ndash ndash ndash

Phase Later prehistoric 93 1358 0398 11628 0001

Historical 208 0 ndash ndash ndash

Soil type Clay 159 0405 0538 0566 0452

Gravel 65 0197 0639 0095 0757

Sand 24 0640 0593 1163 0281

Silt 35 1006 0530 3605 0058

Open 18 0 ndash ndash ndash

Black

Death

NonndashBlack Death 276 1442 0483 8914 0003

Black Death 25 0 ndash ndash ndash

Charnel Non-charnel 213

0067 0389 0030 0862Charnel 88 0 ndash ndash ndash

Age range Neonate 31 2164 0437 24507 lt00005

Child 23 1036 0501 4270 0039

Juvenile 39 0206 0343 0362 0548

Adult 208 0 ndash ndash ndash

Figure 3 The distribution of jittered OHI scores amongst bone samples separated by archaeological phase (1 later pre-historic 2 historical) after specimens from neonatal skeletons anoxic environments and the Black Death cemetery were

removed






The in1047298uence of each of these factors had been anticipated The historical post-neonatal bones

that had not been retrieved from an anoxic or Black Death context were taken to represent those

that had been exposed to consistently high levels of putrefaction We had predicted that bacterial

bioerosion amongst this historical baseline sample set would be consistently high However

there was a signi1047297cant difference between the historical baseline OHI distribution and an ideal-ized normal curve (n = 170 KolmogorovndashSmirnov Z = 2761 p = 0000 Fig S4) The historical

baseline distribution differed signi1047297cantly from a normalized curve by its substantial leptokurtic

shape (Pearsonrsquos measure of kurtosis = 1544 standard error of kurtosis = 0370 ratio = 417)

Leptokurtic distributions are symptomatic of low variance The historical baseline distribution

was signi1047297cantly less variable around scores of 0 than a normal curve It was possible that larger

site assemblages had enacted an inordinate in1047298uence on the baseline historical distribution of

OHI scores However there were no signi1047297cant differences in distributions of OHI scores be-

tween separate historical site assemblages included within the baseline sample set (n =121

KruskalndashWallis Χ 2 = 11402 p = 0122 Fig S5)

The distribution of OHI scores amongst the later prehistoric assemblage deviated signi1047297cantlyfrom a normal distribution when the samples from neonatal skeletons and anoxic environments

were excluded (n = 140 KolmogorovndashSmirnov Z = 1437 p = 0032) The modal OHI score of

the later prehistoric remains was also 0 but the overall distribution displayed a surplus of varia-

tion at scores of 2 and 5 There were signi1047297cant differences in OHI scores between samples of

bone from discrete later prehistoric sites (n = 87 KruskalndashWallis Χ 2 = 24576 p = 0026)

The presence of bacterial tunnelling

Non-Wedl MFD were identi1047297ed within 87 of the study sample The factors that in1047298uenced the

presence of bacterial bioerosion were discerned using binary logistic regression (Table 2) In de-scending order age at death phase and an anoxic environment had affected the presence of bac-

terial bioerosion All signi1047297cant variation in the occurrence of bacterial attack with age at death

was encompassed by the neonatal samples (Fig S6) The elevated distribution of neonatal OHI

scores was solely attributable to the high proportion of samples that were free from bacterial

bioerosion

Higher proportions of later prehistoric bone samples were free from bacterial tunnelling

(Fig S7) Higher proportions of historical bones from anoxic contexts were free from bacterial

bioerosion (Fig S8) When the bone samples from neonatal individuals and anoxic burial environ-

ments were removed there was no statistically signi1047297cant difference in the occurrence of non-

Wedl MFD amongst bones from discrete historical sites (n = 146 Fisher rsquos exact test = 7119

p = 0431) By contrast there were signi1047297cant differences in the appearance of bacterial bioerosion

amongst later prehistoric site assemblages after the same samples were disregarded (n =87

Fisher rsquos exact test = 26104 p = 0001)

DISCUSSION

Bacterial bioerosion and soil type

Neither the extent nor the occurrence of bacterial bioerosion correlated with burial soil Burial

soil was de1047297

ned crudely although a lack of association between histological preservation of ar-chaeological bone and burial sediment has been noted by previous studies (Hanson and Buikstra

1987 Nielsen-Marsh et al 2007 Smith et al 2007) In several instances articulated and

8 T J Booth





disarticulated bones of comparable age that lay in close proximity within similar sediment dem-

onstrated opposing levels of histological preservation The specimens that constituted the histor-

ical baseline assemblage originated from dispersed sites and variable burial sediments yet displayed no signi1047297cant variation in bacterial bioerosion The Carsington Pasture Cave assem-

blage had probably never lain within sediment yet most of the bones demonstrated extensive

levels of bacterial bioerosion (Papakonstantinou 2009)

No samples demonstrated signi1047297cant occurrences of diagenetic features indicative of chemical

corrosion such as micro1047297ssures or generalized destruction (Nielsen-Marsh et al 2007 Hollund

et al 2012) The absence of these features supported the assumption that most sampled bones

originated from benign soils These observations suggested that bacterial bioerosion of archaeo-

logical bones was not linked to features of the burial environment beyond anoxia contrary to

what would be expected had bioerosion been caused by soil bacteria (Jans et al 2004)

Bacterial bioerosion and age at death

The in1047298uence of age at death on bacterial bioerosion was dictated by the dichotomy between neo-

natal and post-neonatal remains This 1047297nding was inconsistent with Turner-Walker rsquos (2008 16)

hypothesis of bacterial bioerosion being controlled by age-related microstructural changes to the

bone which has been used previously to argue against a relationship between bacterial

bioerosion and early taphonomy (Turner-Walker et al 2002 Turner-Walker 2008) The opposi-

tion between neonatal and post-neonatal remains was de1047297ned by the absence of bacterial

bioerosion from the former White and Booth (2014) found that at 1 year post mortem bones

from buried stillborn neonatal piglet carcasses were free from bioerosion whereas those from born neonatal and juvenile pigs had been extensively bioeroded by bacteria This pattern was

most probably linked to the development of the gut microbiome at or soon after birth (Mackie

Table 2 Results of the binary logistic regression analysis applied to the presence of bacterial bioerosion signi 1047297cant

p-values are highlighted in bold

Variable Outcome

Number of

samples B

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)

Anoxia Presentabsent 301 3362 0982 11716 0001

Phase Later prehistoric

historical

301 2792 0771 13121 lt00005

Soil type Overall 301 ndash ndash 5338 0254

Clay 159 0339 0833 0165 0684

Gravel 65 1023 1020 1005 0316

Sand 24 1726 0855 2227 0136

Silt 35 20660 6310078 0000 0997


Black

Death

NonndashBlack Death

Black Death

301 0744 1046 0506 0477

Charnel Non-charnel charnel

301

0948 0881 1159 0282

Age range Overall 301 ndash ndash 24408 lt00005

Neonate 31 4159 0856 23623 lt00005

Child 23 0252 1172 0046 0830

Juvenile 39 0034 0708 0002 0962

Adult 208 0 ndash 0569 0450






et al 1999 White and Booth 2014) The binary preservation of the neonatal archaeological hu-

man bones echoed White and Boothrsquos (2014) results and was consistent with a dichotomy be-

tween infants that had variably lived long enough to develop colonies of osteolytic gut

microbiota The majority (1215) of the unbioeroded neonatal human samples originated from

dispersed historical cemeteries where most samples had been extensively bioeroded These re-sults are inconsistent with a scenario in which bioerosion is caused by exogenous bacteria par-

ticularly when the low mineralization of neonatal bone is likely to increase its susceptibility to

bacterial attack from the soil (Hackett 1981)

Bacterial bioerosion and anoxia

The relationship between anoxic environments putrefaction and bone bioerosion is well docu-

mented and the correlation between histological preservation and anoxia was expected

(Turner-Walker and Jans 2008 Hollund et al 2012 Turner-Walker 2012) Anoxic environments

limited rather than prevented bacterial bone bioerosion This result was probably a consequenceof anoxia at most sites having been caused by intermittent waterlogging Variation in the OHI

scores was likely to be related to how far bodily decomposition had progressed before each grave

had become inundated although episodic waterlogging would have a similar effect on any aer-

obic osteolytic soil bacteria (Turner-Walker and Jans 2008 Hollund et al 2012) Histological

preservation of bones from anoxic environments cannot be used to reliably infer early anthropo-

genic taphonomic treatment beyond the level of bodily decomposition that had occurred before

the environment had become anoxic (Turner-Walker and Jans 2008 Hollund et al 2012)

Bacterial bioerosion and the Black Death cemetery

The evidence that the Black Death remains had decomposed above ground had suggested that theywould demonstrate deviant signatures of bacterial bioerosion compared with the rest of the consis-

tently buried historical assemblage (Grainger et al 2008 19) Unburied bodies would have been

rapidly skeletonized by insects thereby reducing the levels of soft tissue putrefaction that the

bones experienced post-burial (Simmons et al 2010 White and Booth 2014) The variably ele-

vated histological preservation of the Black Death samples was consistent with this model The

partial anatomical articulation of these skeletons suggested that burial had taken place before de-

composition had progressed signi1047297cantly (Grainger et al 2008 19) This observation was consis-

tent with the lack of association between Black Death and the presence of non-Wedl MFD as all

skeletons would have experienced some post-burial soft tissue putrefaction

Bacterial bioerosion and archaeological phase

The signi1047297cant in1047298uence of archaeological phase on bacterial bone bioerosion inferred that there

was a detectable relationship between bacterial attack and funerary treatment The temporal origin

of a bone sample had a larger in1047298uence on the appearance of bacterial bioerosion than anoxia This

result may be explained by the observation that bones that survive well over longer timescales

tend to display well-preserved microstructures (Trueman and Martill 2002) However bone sam-

ples of similar antiquity demonstrated highly variable diagenetic signatures and previous studies

have consistently failed to identify a correlation between chronological age and bacterial attack

(Hedges et al 1995 Hedges 2002) Bacterial bioerosion amongst the historical baseline assem-blage was invariably high consistent with what would be expected within bones of intact bodies

that had been interred soon after death (Hedges 2002 Jans et al 2004 Nielsen-Marsh et al 2007

10 T J Booth





White and Booth 2014) The persistent variation in bacterial bioerosion amongst the later prehis-

toric assemblages was consistent with the knowledge that these individuals were subject to vari-

able early post mortem treatment that exposed the bones to diverse levels of bacterial attack

The dichotomy in OHI scores between historical and later prehistoric samples could be ex-

plained by soil bacteria Most of the historical samples came from large established cemeterieswhich would be more likely to contain bacterial colonies specially adapted to breaking down

bone proteins (Janaway 1987) The rapid removal of soft tissue involved in putative prehistoric

funerary treatments such as excarnation or dismemberment may reduce the attractiveness of bone

to bioerosive soil bacteria (Hedges 2002) However the lack of association between OHI and soil

type as well as the stark results from the neonatal samples were more consistent with an endog-

enous model of bone bioerosion and contradicted expectations related to exogenous bacterial in-

vasion Within the context of the current study the best explanation for the relationship between

archaeological phase and bone diagenesis was that bacterial bioerosion re1047298ected the extent to

which variable phase-speci1047297c funerary processes had exposed the bones to bodily putrefaction

The lack of variation in bacterial bioerosion amongst the historical bone samples suggestedthat universally variable natural and anthropogenic factors that are conventionally thought to af-

fect bodily decomposition had not in1047298uenced bacterial bone bioerosion These factors included

season of death climatic variation within a temperate zone cof 1047297n burial clothingwrapping

burial depth microbiome health and composition diet infectious disease and penetrative trauma

(Rodriguez and Bass 1983 1985 Mant 1987 Mann et al 1990 Campobasso et al 2001 Cross

and Simmons 2010 Vass 2011 Zhou and Bayard 2011 Ferreira and Cunha 2013) The com-

mon feature of these variables is that they affect the rate of bodily decomposition but not the

overall level of putrefaction experienced by the bones (Rodriguez and Bass 1983 1985

Janaway 1996 Rodriguez 1997 Campobasso et al 2001 Vass 2011 Zhou and Bayard 2011

Ferreira and Cunha 2013) Only processes that suf 1047297ciently reduce the level of putrefaction ex-perienced by a bone such as rapid extraneous soft tissue loss will affect bacterial bone

bioerosion (Jans et al 2004 Nielsen-Marsh et al 2007)

The association between histological bone preservation and ancient organic biomolecular yield

(collagen and aDNA) suggests that the results of the current study posit some counter-intuitive

ideas regarding models of biomolecular preservation Neonatal bones may be more likely to re-

tain higher levels of intact organic biomolecules with the caveat that their small size renders

them more susceptible to contamination chemical diagenesis and mechanical weathering Most

discussions of biomolecular preservation are concerned with environmental conditions and time

but the results of our study emphasize the role of cultural practices (Perry et al 1988 Grupe

1995 Geigl 2002 Goumltherstroumlm et al 2002 Haynes et al 2002 Rollo et al 2002 Deviegraveseet al 2010) Temporal variation in funerary practices suggests that in certain cases older bones

may be more viable for organic biomolecular analyses Future studies of biomolecular survival

in archaeological bones could test these assertions Fossilized bones usually demonstrate low

levels of bioerosion and therefore these same factors may also have some in1047298uence on biases

within the fossil record (Trueman and Martill 2002)

CONCLUSIONS

In conclusion the results of our study suggested that early anthropogenic treatment is not the pri-

mary factor that in1047298

uences bacterial bioerosion within archaeological human bones The immac-ulate histological preservation of almost half of neonatal samples inferred that the sterility of

stillborn infant intestinal tracts ensured that their bones were unaffected by bacterial tunnelling






Anoxic burial environments primarily dictated patterns of bacterial bone bioerosion When these

factors were controlled for there was a signi1047297cant relationship between bacterial bioerosion and

archaeological phase the nature of which corresponded with patterns of bodily decomposition

encouraged by known variable phase-speci1047297c funerary treatments The best explanation for these

observations was a predictable relationship between funerary treatment and bacterial bioerosionof bone Measures of putrefactive bacterial bioerosion within archaeological bones should pro-

vide a useful tool for re1047297ning reconstructions of past funerary practices as part of wider holistic

taphonomic analyses These results could be incorporated into predictive models of organic

biomolecular preservation and fossilization

ACKNOWLEDGEMENTS

This research was undertaken as part of an Arts and Humanities Research Council PhD student-

ship (Award No AHI0099571) at the University of Shef 1047297eld I am grateful to my PhD super-

visors Andrew Chamberlain Mike Parker Pearson Pia Nystrom and John Barrett I offer thanks

to those individuals and institutions that facilitated sampling Alison Sheridan (National Museum

of Scotland) Geoff Morley (MOLES Archaeology) Paul Wilkinson (Swale and Thames Archae-

ological Survey Company) Mark Knight (Cambridge Archaeological Unit) Olivia Lelong

(Northlight Heritage) Richard Madgwick (University of Cardiff) Dave Allen (Hampshire

Museum Service) Justyna Miszkiewicz and Patrick Mahoney (University of Kent) Peter Rowe

(Tees Archaeology) and Karl-Goumlran Sjoumlgren (University of Gothenburg) I also thank Ian

Barnes Matthew Collins and Selina Brace for commenting on early versions of this paper

REFERENCES

Balzer A Gleixner G Grupe G Schmidt H L Schramm S and Turban-Just S 1997 In vitro decomposition of

bone collagen by soil bacteria the implications for stable isotope analysis in archaeometry Archaeometry 39

415ndash29

Bass W 1995 Human osteology a laboratory and 1047297eld manual 4th edn Missouri Archaeological Publications

Columbia OH

Bell L S Skinner M F and Jones S J 1996 The speed of post mortem change to the human skeleton and its

taphonomic signi1047297cance Forensic Science International 82 129ndash40

Bethel P H and Carver M O H 1987 Detection and enhancement of decayed inhumations at Sutton Hoo in Death

decay and reconstruction approaches to archaeology and forensic science (eds A Boddington A N Garland and

R C Janaway) pp 10ndash21 Manchester University Press Manchester

Breitmeier D Graefe-Kirci U Albrecht K Weber M Troumlger H D and Kleemann W J 2005 Evaluation of the

correlation between time corpses spent in in-ground graves and 1047297ndings at exhumation Forensic Science Interna-tional 154 218ndash23

Brickley M and McKinley J I 2004 Guidelines to the standards for recording human remains BABAO and IFA

Paper No 7

Buikstra J E and Ubelaker D H 1994 Standards for data collection from human skeletal remains Arkansas

Archeological Survey Fayetteville AR

Campobasso C P Giancarlo D V and Introna F 2001 Factors affecting decomposition and Diptera colonization

Forensic Science International 120 18ndash27

Carter D O Yellowlees D and Tibbett M 2008 Temperature affects microbial decomposition of cadavers ( Rattus

rattus) in contrasting soils Applied Soil Ecology 40 129ndash37

Carter D O Yellowlees D and Tibbett M 2010 Moisture can be the dominant environmental parameter governing

cadaver decomposition in soil Forensic Science International 200 60ndash6

Child A M 1995a Microbial taphonomy of archaeological bone Studies in Conservation 40(1) 19ndash30Child A M 1995b Towards an understanding of the microbial decomposition of archaeological bone in the burial

environment Journal of Archaeological Science 22 165ndash74

12 T J Booth





Cipollaro M Di Bernardo G Galano G Galderisi U Guarino F Angelini F and Cascino A 1998 Ancient DNA

in human bone remains from Pompeii archaeological site Biochemical and Biophysical Research Communications

247 901ndash4

Collins M J Penkman K E Rohland N Shapiro B Dobberstein R C Ritz-Timme S and Hofreiter M 2009 Is

amino acid racemization a useful tool for screening for ancient DNA in bone Proceedings of the Royal Society B

Biological Sciences 276(1669) 2971ndash7Colson I B Bailey J F Vercauteren M Sykes B C and Hedges R E M 1997 The preservation of ancient DNA

and bone diagenesis Ancient Biomolecules 1(2) 109ndash17

Cotton G E Aufderheide A C and Goldschmidt V G 1987 Preservation of human tissue immersed for 1047297ve years in

fresh water of known temperature Journal of Forensic Sciences 32(4) 1125ndash30

Cox M and Mays S 2000 Human osteology in archaeology and forensic science Greenwich Medical Media London

Cross P and Simmons T 2010 The in1047298uence of penetrative trauma on the rate of decomposition Journal of Forensic

Sciences 55(2) 295ndash301

Daniell C 1997 Death and burial in medieval England 1066 ndash 1550 Routledge London

Darvill T 2010 Prehistoric Britain Routledge London

Dent B B Forbes S L and Stuart B H 2004 Review of human decomposition processes in soil Environmental

Geology 45 576ndash85

Deviegravese T Colombini M P Regert M Stuart B H and Guerbois J P 2010 TGMS analysis of archaeologicalbone from burials of the late Roman period Journal of Thermal Analysis and Calorimetry 99 811ndash13

Dixon R A Dawson L and Taylor D 2008 The experimental degradation of archaeological human bone by

anaerobic bacteria and the implications for the recovery of Ancient DNA in The proceedings of the 9th International

Conference on Ancient DNA and Associated Biomolecules 19 ndash 22 October 2008 Pompeii Italy

Economou C 2003 Behind the north wall of sleep microbial degradation of foetal and neonatal bone with a case study

from Bolsover Unpublished MSc dissertation University of Shef 1047297eld

Fernaacutendez-Jalvo Y Andrews P Pesquero D Smith C Mariacuten-Monfort D Saacutenchez B Geigl E-M and Alonso

A 2010 Early bone diagenesis in temperate environments part I surface features and histology Palaeogeography

Palaeoclimatology Palaeoecology 288 62ndash81

Ferreira M T and Cunha E 2013 Can we infer post mortem interval on the basis of decomposition rate A case from a

Portuguese cemetery Forensic Science International 226(1) 298e1ndash6

Fielder S and Graw M 2003 Decomposition of buried corpses with special reference to the formation of adipocere Naturwissenschaft 90 291ndash300

Geigl E-M 2002 On the circumstances surrounding the preservation and analysis of very old DNA Archaeometry 44

337ndash42

Gill-King H 1997 Chemical and ultrastructural aspects of decomposition in Forensic taphonomy the postmortem fate

of human remains (eds W D Haglund and M H Sorg) pp 93 ndash108 CRC Press Boca Raton FL

Gordon C C and Buikstra J E 1981 Soil pH bone preservation and sampling bias at mortuary sites American

Antiquity 46(3) 566ndash71

Goumltherstroumlm A Collins M J Angerbjoumlrn A and Lideacuten K 2002 Bone preservation and DNA ampli1047297cation

Archaeometry 44 395ndash404

Grainger I Hawkins D Cowal L and Mikulski R 2008 The Black Death cemetery East Smith 1047297eld London

Archaeology Monograph 43 Museum of London London

Grupe G 1995 Preservation of collagen in bone from dry sandy soil Journal of Archaeological Science 22 193ndash

9Grupe G and Dreses-Werringloer U 1993 Decomposition phenomena in thin-sections of excavated human bones in

Histology of ancient human bone methods and diagnosis (eds G Grupe and A N Garland) pp 27ndash36 Springer-

Verlag Berlin

Grupe G and Turban-Just S 1998 Serum proteins in archaeological human bone International Journal of

Osteoarchaeology 6(3) 300ndash8

Guarino F M Angelini F Vollono C and Ore1047297ce C 2006 Bone preservation in human remains from the Terme del Sarno

at Pompeii using light microscopy and scanning electron microscopy Journal of Archaeological Science 33 513ndash20

Hackett C J 1981 Microscopical focal destruction (tunnels) in exhumed human bones Medicine Science and the Law

21(4) 243ndash66

Hagelberg E Bell L S Allen T Boyde A Jones S J Clegg J B Hummel S Brown T A and Ambler R P

1991 Analysis of ancient bone DNA techniques and applications (and discussion) Philosophical Transactions of the

Royal Society B Biological Sciences 333

(1268) 399ndash

407Hanson D B and Buikstra J E 1987 Histomorphological alteration in buried human bone from the Lower Illinois

Valley implications for palaeodietary research Journal of Archaeological Science 14 549ndash63






Haynes S Searle J B Bretman A and Dobney K M 2002 Bone preservation and ancient DNA the application of

screening methods for predicting DNA survival Journal of Archaeological Science 29(6) 585ndash92

Hedges R E M 2002 Bone diagenesis an overview of processes Archaeometry 44 319ndash28

Hedges R E M Millard A R and Pike A W G 1995 Measurements and relationships of diagenetic alteration of

bone from three archaeological sites Journal of Archaeological Science 22 201ndash9

Hollund H I Jans M M E Collins M J Kars H Joosten I and Kars S M 2012 What happened here Bonehistology as a tool in decoding the postmortem histories of archaeological bone from Castricum The Netherlands

International Journal of Osteoarchaeology 22(5) 537ndash48

Jackes M Sherburne R Lubell D Barker C and Wayman M 2001 Destruction of microstructure in archaeological

bone a case study from Portugal International Journal of Osteoarchaeology 11 415ndash32

Jagger K A and Rogers T L 2009 The effects of soil environment on postmortem interval a macroscopic analysis

Journal of Forensic Sciences 54(6) 1217ndash21

Jakobsson H E Abrahamsson T R Jenmalm M C Harris K Quince C Jernberg C Bjoumlrksteacuten B Engstrand L

and Andersson A F 2013 Decreased gut microbiota diversity delayed Bacterioidetes colonisation and reduced Th1

responses in infants delivered by Caesarean section Gut 63 559ndash66

Janaway R C 1987 The preservation of organic materials in association with metal artefacts deposited in inhumation

graves in Death decay and reconstruction approaches to archaeology and forensic science (eds A Boddington

A N Garland and R C Janaway) pp 109 ndash26 Manchester University Press ManchesterJanaway R C 1996 The decay of buried human remains and their associated material in Studies in crime an introduc-

tion to forensic archaeology (eds J Hunter C Roberts and A Martin) pp 58 ndash85 Batsford London

Jans M M E Nielsen-Marsh C M Smith C I Collins M J and Kars H 2004 Characterisation of microbial attack

on archaeological bone Journal of Archaeological Science 31 87ndash95

Lee-Thorp J A and Sealy J C 2008 Beyond documenting diagenesis the Fifth International Bone Diagenesis

Workshop Palaeogeography Palaeoclimatology Palaeoecology 266 129ndash33

Mackie R I Sghir A and Gaskins H R 1999 Developmental microbial ecology of the neonatal gastrointestinal tract

American Journal of Clinical Nutrition 69(suppl) 1035ndash45

Manhein M H 1997 Decomposition rates of deliberate burials a case study of preservation in Forensic taphonomy

the postmortem fate of human remains (eds W D Haglund and M H Sorg) pp 469ndash81 CRC Press Boca

Raton FL

Mann R W Bass W M and Meadows L 1990 Time since death and decomposition of the human body variablesand observations in case and experimental 1047297eld studies Journal of Forensic Sciences 35(1) 103ndash11

Mant A K 1987 Knowledge acquired from post-war exhumations in Death decay and reconstruction approaches to

archaeology and forensic science (eds A Boddington A N Garland and R C Janaway) pp 65ndash78 Manchester

University Press Manchester

Marchiafava V Bonucci E and Ascenzi A 1974 Fungal osteoclasia a model of dead bone resorption Calci 1047297ed

Tissue Research 14 195ndash210

Meyer J Anderson B and Carter D O 2013 Seasonal variation of carcass decomposition and gravesoil chemistry in

a cold (Dfa) climate Journal of Forensic Sciences 58(5) 1175ndash82

Millard A 2001 The deterioration of bone in Handbook of archaeological sciences (eds D Brothwell and A M Pollard)

pp 637ndash47 Wiley Chichester

Nielsen-Marsh C M and Hedges R E M 2000 Patterns of diagenesis in bone I the effects of site environments

Journal of Archaeological Science 27 1139ndash

50Nielsen-Marsh C M Smith C I Jans M M E Nord A Kars H and Collins M J 2007 Bone diagenesis in the

European Holocene II taphonomic and environmental considerations Journal of Archaeological Science 34(9)

1523ndash31

OrsquoConnor S Ali E Al-Sabah S Anwar D Bergstroumlm E Brown K A Buckberry J Buckley S Collins M

Denton J Dorling K Dowle A Duffey P Edwards H G M Correia Faria E Gardner P Gledhill A

Heaton K Heron C Janaway R C Keely B J King D Masinton A Penkman K Petzold A Pickering

M D Rumsby M Schutkowski H Shackleton K A Thomas J Thomas-Oates J Usai M-R Wilson A S

and OrsquoConnor T 2011 Exceptional preservation of a prehistoric human brain from Heslington Yorkshire UK

Journal of Archaeological Science 38 1641ndash54

Ottoni C Koon H E Collins M J Penkman K E Rickards O and Craig O E 2009 Preservation of ancient

DNA in thermally damaged archaeological bone Naturwissenschaften 96(2) 267ndash78

Papakonstantinou N 2009 Human skeletal remains from Neolithic caves in the Peak District an osteoarchaeological and taphonomic approach Unpublished MSc dissertation University of Shef 1047297eld

Parker Pearson M 1999 The archaeology of death and burial Sutton Stroud

14 T J Booth





Parker Pearson M Chamberlain A Craig O Marshall P Mulville J Smith J Chenery C Collins M Cook G

Craig G Evans J Hiller J Montgomery J Schwenninger J-L Taylor G and Wess T 2005 Evidence for

mummi1047297cation in Bronze Age Britain Antiquity 79 529ndash46

Perry W L Bass W M Riggsby W S and Sirotkin K 1988 The autodegradation of deoxyribonucleic acid (DNA)

in human rib bone and its relationship to the time interval since death Journal of Forensic Sciences 33(1) 144ndash53

Piepenbrink H 1986 Two examples of biogenous dead bone decomposition and their consequences for taphonomicinterpretation Journal of Archaeological Science 13 417ndash30

Piepenbrink H 1989 Examples of chemical change during fossilisation Applied Geochemistry 4 273ndash80

Polson C J Gee D J and Knight B 1985 The essentials of forensic medicine Pergamon Press Oxford

Redfern R 2008 New evidence for Iron Age secondary burial practice and bone modi1047297cation from Gussage All Saints

and Maiden Castle (Dorset England) Oxford Journal of Archaeology 27(3) 281ndash301

Rodriguez W C 1997 Decomposition of buried and submerged bodies in Forensic taphonomy the postmortem fate of

human remains (eds W D Haglund and M H Sorg) pp 93 ndash108 CRC Press Boca Raton FL

Rodriguez W C and Bass W M 1983 Insect activity and its relationship to decay rates of human cadavers in East

Tennessee Journal of Forensic Sciences 28(2) 423ndash32

Rodriguez W C and Bass W M 1985 Decomposition of buried bodies and methods that may aid in their location


Rollo F Ubaldi M Marota I Luciani S and Ermini L 2002 DNA diagenesis effects of environment and time onhuman bone Ancient Biomolecules 4(1) 1ndash7

Scheuer J L and Black S 2000 Development and aging of the juvenile skeleton in Human osteology in archaeology

and forensic science (eds M Cox and S Mays) pp 9ndash23 Greenwich Medical Media London

Schwartz J 1995 Skeleton keys an introduction to human skeletal morphology development and analysis Oxford

University Press Oxford

Sharon I Morowitz M J Thomas B C Costello E K Relman D A and Ban1047297eld J F 2013 Time series com-

munity genomics analysis reveals rapid shifts in bacterial species strains and phage during infant gut colonization

Genome Research 23 111ndash20

Simmons T Cross P A Adlam R E and Moffatt C 2010 The in1047298uence of insects on decomposition rate in buried

and surface remains Journal of Forensic Sciences 55(4) 889ndash92

Smith C I Nielsen-Marsh C M Jans M M E and Collins M J 2007 Bone diagenesis in the European Holocene I

patterns and mechanisms Journal of Archaeological Science 34(9) 1485ndash

93Trueman C N and Martill D M 2002 The long-term survival of bone the role of bioerosion Archaeometry 44 371ndash82

Turner B and Wiltshire P 1999 Experimental validation of forensic evidence a study of the decomposition of buried

pigs in a heavy clay soil Forensic Science International 101 113ndash22

Turner-Walker G 2008 The chemical and microbial degradation of bones and teeth in Advances in human

palaeopathology (eds R Pinhasi and S Mays) pp 3ndash30 Wiley Chichester

Turner-Walker G 2012 Early bioerosion in skeletal tissues persistence through deep time Neues Jahrbuch fuumlr

Geologie und Palaumlontologie 265 165ndash83

Turner-Walker G and Jans M 2008 Reconstructing taphonomic histories using histological analysis

Palaeogeography Palaeoclimatology Palaeoecology 266 227ndash35

Turner-Walker G and Syversen U 2002 Quantifying histological changes in archaeological bones using BSEndashSEM

image analysis Archaeometry 44 461ndash8

Turner-Walker G Nielsen-Marsh C M Syversen U Kars H and Collins M J 2002 Sub-micron spongiform porosity is the major ultra-structural alteration occurring in archaeological bone International Journal of

Osteoarchaeology 12 407ndash14

Vass A A 2011 The elusive universal post-mortem interval formula Forensic Science International 204 34ndash40

Vranyacute B Hnaacutetkovaacute Z and Lettl A 1988 Occurrence of collagen-degrading microorganisms in associations of

mesophilic heterotrophic bacteria from various soils Folia Microbiologica 33 458ndash61

White L and Booth T J 2014 The origin of bacteria responsible for bioerosion to the internal bone microstructure

results from experimentally-deposited pig carcasses Forensic Science International 239 92ndash102

Wilson A S Janaway R C Holland A D Dodson H I Baran E Pollard A M and Tobin D J 2007 Modelling

the buried human body environment in upland climes using three contrasting 1047297eld sites Forensic Science Interna-

tional 169 6ndash18

Yoshino M Kimijima T Miyasaka S Sato H and Seta S 1991 Microscopic study on estimation of time since

death in skeletal remains Forensic Science International 49

143ndash

58Zhou C and Bayard R W 2011 Factors and processes causing accelerated decomposition in human cadaversmdashan

overview Journal of Forensic and Legal Medicine 18 6ndash9






SUPPORTING INFORMATION

Additional Supporting Information may be found in the online version of this article at the

publisher rsquos website

Table S1 Catalogue of archaeological sites included in this study

Figure S1 Distribution of jittered OHI scores amongst samples separated by age at death

(1 = Neonate 2 = Child 3 = Juvenile 4 = Adult)

Figure S2 Distribution of jittered OHI scores amongst samples of bone variably retrieved from

anoxic environments (0 = anoxia was absent 1 = anoxia was present) after the neonatal specimens

had been excluded

Figure S3 Distribution of jittered OHI scores amongst samples of bone that had been variably re-

trieved from the Black Death cemetery (0 = Non-Black Death grave 1 = Black Death grave) after

specimens from neonatal skeletons and anoxic environments had been removed

Figure S4 Distribution of OHI scores amongst the historical rsquobaselinersquo assemblage when all non-

zero scores were replicated as negative numbers with a superimposed normal curve

Figure S5 Distribution of OHI scores amongst the samples that constituted the historical baseline

assemblage separated by site

Figure S6 Proportions of samples affected by bacterial tunnelling separated by age-at-death

Figure S7 Proportions of samples affected by bacterial tunnelling separated by archaeological

phase after the neonatal specimens had been excluded

Figure S8 Proportions of historical samples that demonstrated bacterial tunnelling separated by

whether they had been retrieved from an anoxic environment after the neonatal samples had

been excluded

16 T J Booth





Early theories regarding the aetiology of bacterial bone bioerosion were concerned with soil

micro-organisms (Marchiafava et al 1974 Hackett 1981 Piepenbrink 1986 1989 Hanson

and Buikstra 1987 Yoshino et al 1991 Grupe and Dreses-Werringloer 1993) Successive

studies have suggested that non-Wedl MFD are produced by a dead organism rsquos gut bacteria (Child

1995a Bell et al 1996 Jans et al 2004 Guarino et al 2006 Nielsen-Marsh et al 2007 Hollundet al 2012 White and Booth 2014) The post mortem deterioration of an organism rsquos immune sys-

tem and mucosal membranes facilitate the transmigration of gut bacteria within the 1047297rst few days

after death (Polson et al 1985 Janaway 1987 Child 1995ab Gill-King 1997) These micro-

organisms are primarily responsible for bodily putrefaction (Polson et al 1985 Janaway 1987

Child 1995ab) An endogenous origin of osteolytic microbiota suggests that bacterial bone

bioerosion should re1047298ect the extent to which the skeleton was exposed to putrefaction

This hypothesis presents a potentially useful prospect for the study of funerary archaeology

Different post mortem treatments should expose the skeleton to divergent levels of putrefaction

and leave characteristic signatures of bacterial bioerosion within the bone microstructure (Bell

et al 1996 Jans et al 2004 Parker Pearson et al 2005 Hollund et al 2012 White and Booth2014) Certain studies have already detected logical relationships between bacterial bone

bioerosion and early taphonomic events (Bell et al 1996 Jans et al 2004 Nielsen-Marsh

et al 2007 White and Booth 2014)

Measures of bacterial bioerosion in archaeological human bone combined with models of de-

composition could help to distinguish between discrete funerary rites that leave similar archaeo-

logical records or help to identify hidden complexities in funerary treatment (Bell et al 1996

Jans et al 2004 Parker Pearson et al 2005 Nielsen-Marsh et al 2007 Smith et al 2007

Turner-Walker and Jans 2008 Hollund et al 2012) Furthermore microbial exploitation is a pri-

mary mechanism of bone collagen loss and has been variably linked with successful extraction of

organic biomolecules including DNA (Hagelberg et al 1991 Grupe 1995 Colson et al 1997Cipollaro et al 1998 Geigl 2002 Goumltherstroumlm et al 2002 Haynes et al 2002 Rollo et al

2002 Collins et al 2009 Ottoni et al 2009 Deviegravese et al 2010) Establishing the factors that

affect bone bioerosion may help to account for variation in organic biomolecular yield amongst

archaeological bones and contribute to predictive models of preservation

There have been no systematic investigations into whether speci1047297c funerary treatments pro-

duce predictable signatures of bacterial bioerosion within archaeological human remains It has

yet to be determined whether the effects of anthropogenic processes can be distinguished from

changes elicited by the burial environment Seasonality is often cited as the most signi1047297cant fac-

tor that affects the nature of bodily decomposition (Rodriguez and Bass 1983 1985 Mann et al

1990 Manhein 1997 Campobasso et al 2001 Wilson et al 2007 Zhou and Bayard 2011Meyer et al 2013) Anoxic burial environments can arrest bodily putrefaction and have been

consistently observed to discourage bacterial bioerosion of bone microstructures (Polson et al

1985 Cotton et al 1987 Mant 1987 Janaway 1996 Turner and Wiltshire 1999 Fielder and

Graw 2003 Wilson et al 2007 Turner-Walker and Jans 2008 OrsquoConnor et al 2011 Hollund

et al 2012) The endogenous model of bacterial bioerosion and the in1047298uence of early taphonomic

events have been questioned more recently (Turner-Walker 2008 2012 Fernaacutendez-Jalvo et al

2010) The strength of the relationship between early post mortem processes and bacterial bone

bioerosion is questionable and needs to be explored further to determine how it might be used in

reconstructions of funerary activities in the past

In order to establish the relationship between bacterial bone bioerosion and funerary treat-ment and to assess whether microscopic analysis of archaeological human remains could aid

reconstructions of funerary treatments practised by past populations we conducted

2 T J Booth





















decomposition






Materials









































cantly in1047298





4 T J Booth







































and Booth 2014)







possibility






RESULTS


























6 T J Booth










highlighted in bold

Variable Outcome

Number of

samples

Parameter

estimate

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)

Anoxia Absent 261 2065 0367 31732 lt00005




Soil type Clay 159 0405 0538 0566 0452

Gravel 65 0197 0639 0095 0757

Sand 24 0640 0593 1163 0281

Silt 35 1006 0530 3605 0058


Black

Death






Child 23 1036 0501 4270 0039

Juvenile 39 0206 0343 0362 0548



removed






























bioerosion









DISCUSSION



soil was de1047297



8 T J Booth




























Variable Outcome

Number of

samples B

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)



historical

301 2792 0771 13121 lt00005


Clay 159 0339 0833 0165 0684

Gravel 65 1023 1020 1005 0316

Sand 24 1726 0855 2227 0136

Silt 35 20660 6310078 0000 0997


Black

Death

NonndashBlack Death

Black Death

301 0744 1046 0506 0477


301

0948 0881 1159 0282


Neonate 31 4159 0856 23623 lt00005

Child 23 0252 1172 0046 0830

Juvenile 39 0034 0708 0002 0962

Adult 208 0 ndash 0569 0450












































10 T J Booth









































CONCLUSIONS


















ACKNOWLEDGEMENTS











REFERENCES



415ndash29


Columbia OH









Paper No 7











12 T J Booth







247 901ndash4



























337ndash42












Verlag Berlin






21(4) 243ndash66




(1268) 399ndash


































Raton FL















1523ndash31











14 T J Booth





















































tional 169 6ndash18



143ndash
















had been excluded













been excluded

16 T J Booth





















decomposition






Materials









































cantly in1047298





4 T J Booth







































and Booth 2014)







possibility






RESULTS


























6 T J Booth










highlighted in bold

Variable Outcome

Number of

samples

Parameter

estimate

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)

Anoxia Absent 261 2065 0367 31732 lt00005




Soil type Clay 159 0405 0538 0566 0452

Gravel 65 0197 0639 0095 0757

Sand 24 0640 0593 1163 0281

Silt 35 1006 0530 3605 0058


Black

Death






Child 23 1036 0501 4270 0039

Juvenile 39 0206 0343 0362 0548



removed






























bioerosion









DISCUSSION



soil was de1047297



8 T J Booth




























Variable Outcome

Number of

samples B

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)



historical

301 2792 0771 13121 lt00005


Clay 159 0339 0833 0165 0684

Gravel 65 1023 1020 1005 0316

Sand 24 1726 0855 2227 0136

Silt 35 20660 6310078 0000 0997


Black

Death

NonndashBlack Death

Black Death

301 0744 1046 0506 0477


301

0948 0881 1159 0282


Neonate 31 4159 0856 23623 lt00005

Child 23 0252 1172 0046 0830

Juvenile 39 0034 0708 0002 0962

Adult 208 0 ndash 0569 0450












































10 T J Booth









































CONCLUSIONS


















ACKNOWLEDGEMENTS











REFERENCES



415ndash29


Columbia OH









Paper No 7











12 T J Booth







247 901ndash4



























337ndash42












Verlag Berlin






21(4) 243ndash66




(1268) 399ndash


































Raton FL















1523ndash31











14 T J Booth





















































tional 169 6ndash18



143ndash
















had been excluded













been excluded

16 T J Booth























cantly in1047298





4 T J Booth







































and Booth 2014)







possibility






RESULTS


























6 T J Booth










highlighted in bold

Variable Outcome

Number of

samples

Parameter

estimate

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)

Anoxia Absent 261 2065 0367 31732 lt00005




Soil type Clay 159 0405 0538 0566 0452

Gravel 65 0197 0639 0095 0757

Sand 24 0640 0593 1163 0281

Silt 35 1006 0530 3605 0058


Black

Death






Child 23 1036 0501 4270 0039

Juvenile 39 0206 0343 0362 0548



removed






























bioerosion









DISCUSSION



soil was de1047297



8 T J Booth




























Variable Outcome

Number of

samples B

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)



historical

301 2792 0771 13121 lt00005


Clay 159 0339 0833 0165 0684

Gravel 65 1023 1020 1005 0316

Sand 24 1726 0855 2227 0136

Silt 35 20660 6310078 0000 0997


Black

Death

NonndashBlack Death

Black Death

301 0744 1046 0506 0477


301

0948 0881 1159 0282


Neonate 31 4159 0856 23623 lt00005

Child 23 0252 1172 0046 0830

Juvenile 39 0034 0708 0002 0962

Adult 208 0 ndash 0569 0450












































10 T J Booth









































CONCLUSIONS


















ACKNOWLEDGEMENTS











REFERENCES



415ndash29


Columbia OH









Paper No 7











12 T J Booth







247 901ndash4



























337ndash42












Verlag Berlin






21(4) 243ndash66




(1268) 399ndash


































Raton FL















1523ndash31











14 T J Booth





















































tional 169 6ndash18



143ndash
















had been excluded













been excluded

16 T J Booth







































and Booth 2014)







possibility






RESULTS


























6 T J Booth










highlighted in bold

Variable Outcome

Number of

samples

Parameter

estimate

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)

Anoxia Absent 261 2065 0367 31732 lt00005




Soil type Clay 159 0405 0538 0566 0452

Gravel 65 0197 0639 0095 0757

Sand 24 0640 0593 1163 0281

Silt 35 1006 0530 3605 0058


Black

Death






Child 23 1036 0501 4270 0039

Juvenile 39 0206 0343 0362 0548



removed






























bioerosion









DISCUSSION



soil was de1047297



8 T J Booth




























Variable Outcome

Number of

samples B

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)



historical

301 2792 0771 13121 lt00005


Clay 159 0339 0833 0165 0684

Gravel 65 1023 1020 1005 0316

Sand 24 1726 0855 2227 0136

Silt 35 20660 6310078 0000 0997


Black

Death

NonndashBlack Death

Black Death

301 0744 1046 0506 0477


301

0948 0881 1159 0282


Neonate 31 4159 0856 23623 lt00005

Child 23 0252 1172 0046 0830

Juvenile 39 0034 0708 0002 0962

Adult 208 0 ndash 0569 0450












































10 T J Booth









































CONCLUSIONS


















ACKNOWLEDGEMENTS











REFERENCES



415ndash29


Columbia OH









Paper No 7











12 T J Booth







247 901ndash4



























337ndash42












Verlag Berlin






21(4) 243ndash66




(1268) 399ndash


































Raton FL















1523ndash31











14 T J Booth





















































tional 169 6ndash18



143ndash
















had been excluded













been excluded

16 T J Booth





RESULTS


























6 T J Booth










highlighted in bold

Variable Outcome

Number of

samples

Parameter

estimate

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)

Anoxia Absent 261 2065 0367 31732 lt00005




Soil type Clay 159 0405 0538 0566 0452

Gravel 65 0197 0639 0095 0757

Sand 24 0640 0593 1163 0281

Silt 35 1006 0530 3605 0058


Black

Death






Child 23 1036 0501 4270 0039

Juvenile 39 0206 0343 0362 0548



removed






























bioerosion









DISCUSSION



soil was de1047297



8 T J Booth




























Variable Outcome

Number of

samples B

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)



historical

301 2792 0771 13121 lt00005


Clay 159 0339 0833 0165 0684

Gravel 65 1023 1020 1005 0316

Sand 24 1726 0855 2227 0136

Silt 35 20660 6310078 0000 0997


Black

Death

NonndashBlack Death

Black Death

301 0744 1046 0506 0477


301

0948 0881 1159 0282


Neonate 31 4159 0856 23623 lt00005

Child 23 0252 1172 0046 0830

Juvenile 39 0034 0708 0002 0962

Adult 208 0 ndash 0569 0450












































10 T J Booth









































CONCLUSIONS


















ACKNOWLEDGEMENTS











REFERENCES



415ndash29


Columbia OH









Paper No 7











12 T J Booth







247 901ndash4



























337ndash42












Verlag Berlin






21(4) 243ndash66




(1268) 399ndash


































Raton FL















1523ndash31











14 T J Booth





















































tional 169 6ndash18



143ndash
















had been excluded













been excluded

16 T J Booth










highlighted in bold

Variable Outcome

Number of

samples

Parameter

estimate

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)

Anoxia Absent 261 2065 0367 31732 lt00005




Soil type Clay 159 0405 0538 0566 0452

Gravel 65 0197 0639 0095 0757

Sand 24 0640 0593 1163 0281

Silt 35 1006 0530 3605 0058


Black

Death






Child 23 1036 0501 4270 0039

Juvenile 39 0206 0343 0362 0548



removed






























bioerosion









DISCUSSION



soil was de1047297



8 T J Booth




























Variable Outcome

Number of

samples B

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)



historical

301 2792 0771 13121 lt00005


Clay 159 0339 0833 0165 0684

Gravel 65 1023 1020 1005 0316

Sand 24 1726 0855 2227 0136

Silt 35 20660 6310078 0000 0997


Black

Death

NonndashBlack Death

Black Death

301 0744 1046 0506 0477


301

0948 0881 1159 0282


Neonate 31 4159 0856 23623 lt00005

Child 23 0252 1172 0046 0830

Juvenile 39 0034 0708 0002 0962

Adult 208 0 ndash 0569 0450












































10 T J Booth









































CONCLUSIONS


















ACKNOWLEDGEMENTS











REFERENCES



415ndash29


Columbia OH









Paper No 7











12 T J Booth







247 901ndash4



























337ndash42












Verlag Berlin






21(4) 243ndash66




(1268) 399ndash


































Raton FL















1523ndash31











14 T J Booth





















































tional 169 6ndash18



143ndash
















had been excluded













been excluded

16 T J Booth





























bioerosion









DISCUSSION



soil was de1047297



8 T J Booth




























Variable Outcome

Number of

samples B

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)



historical

301 2792 0771 13121 lt00005


Clay 159 0339 0833 0165 0684

Gravel 65 1023 1020 1005 0316

Sand 24 1726 0855 2227 0136

Silt 35 20660 6310078 0000 0997


Black

Death

NonndashBlack Death

Black Death

301 0744 1046 0506 0477


301

0948 0881 1159 0282


Neonate 31 4159 0856 23623 lt00005

Child 23 0252 1172 0046 0830

Juvenile 39 0034 0708 0002 0962

Adult 208 0 ndash 0569 0450












































10 T J Booth









































CONCLUSIONS


















ACKNOWLEDGEMENTS











REFERENCES



415ndash29


Columbia OH









Paper No 7











12 T J Booth







247 901ndash4



























337ndash42












Verlag Berlin






21(4) 243ndash66




(1268) 399ndash


































Raton FL















1523ndash31











14 T J Booth





















































tional 169 6ndash18



143ndash
















had been excluded













been excluded

16 T J Booth




























Variable Outcome

Number of

samples B

Standard

error

Wald

X 2

Signi 1047297cance

( p-value)



historical

301 2792 0771 13121 lt00005


Clay 159 0339 0833 0165 0684

Gravel 65 1023 1020 1005 0316

Sand 24 1726 0855 2227 0136

Silt 35 20660 6310078 0000 0997


Black

Death

NonndashBlack Death

Black Death

301 0744 1046 0506 0477


301

0948 0881 1159 0282


Neonate 31 4159 0856 23623 lt00005

Child 23 0252 1172 0046 0830

Juvenile 39 0034 0708 0002 0962

Adult 208 0 ndash 0569 0450












































10 T J Booth









































CONCLUSIONS


















ACKNOWLEDGEMENTS











REFERENCES



415ndash29


Columbia OH









Paper No 7











12 T J Booth







247 901ndash4



























337ndash42












Verlag Berlin






21(4) 243ndash66




(1268) 399ndash


































Raton FL















1523ndash31











14 T J Booth





















































tional 169 6ndash18



143ndash
















had been excluded













been excluded

16 T J Booth











































10 T J Booth









































CONCLUSIONS


















ACKNOWLEDGEMENTS











REFERENCES



415ndash29


Columbia OH









Paper No 7











12 T J Booth







247 901ndash4



























337ndash42












Verlag Berlin






21(4) 243ndash66




(1268) 399ndash


































Raton FL















1523ndash31











14 T J Booth





















































tional 169 6ndash18



143ndash
















had been excluded













been excluded

16 T J Booth









































CONCLUSIONS


















ACKNOWLEDGEMENTS











REFERENCES



415ndash29


Columbia OH









Paper No 7











12 T J Booth







247 901ndash4



























337ndash42












Verlag Berlin






21(4) 243ndash66




(1268) 399ndash


































Raton FL















1523ndash31











14 T J Booth





















































tional 169 6ndash18



143ndash
















had been excluded













been excluded

16 T J Booth













ACKNOWLEDGEMENTS











REFERENCES



415ndash29


Columbia OH









Paper No 7











12 T J Booth







247 901ndash4



























337ndash42












Verlag Berlin






21(4) 243ndash66




(1268) 399ndash


































Raton FL















1523ndash31











14 T J Booth





















































tional 169 6ndash18



143ndash
















had been excluded













been excluded

16 T J Booth







247 901ndash4



























337ndash42












Verlag Berlin






21(4) 243ndash66




(1268) 399ndash


































Raton FL















1523ndash31











14 T J Booth





















































tional 169 6ndash18



143ndash
















had been excluded













been excluded

16 T J Booth































Raton FL















1523ndash31











14 T J Booth





















































tional 169 6ndash18



143ndash
















had been excluded













been excluded

16 T J Booth





















































tional 169 6ndash18



143ndash
















had been excluded













been excluded

16 T J Booth













had been excluded













been excluded

16 T J Booth



Booth 2015 Archaeometry

Documents

Transcript of Booth 2015 Archaeometry