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i
Recent Advances in Ageing
and Sexing Animal Bones

For Vasili and Marilena

Recent Advances in Ageing
and Sexing Animal Bones
Proceedings of the 9th Conference of the International Council
of Archaeozoology, Durham, August 2002
Series Editors: Umberto Albarella, Keith Dobney and Peter Rowley-Conwy
Edited by
Deborah Ruscillo
Oxbow Books

Published by
Oxbow Books, Park End Place, Oxford OX1 1HN
© Oxbow Books and the individual authors 2006
ISBN 978 1 842 17122 6 1 84217 122 4
A CIP record for this book is available from The British Library
This book is available direct from
Oxbow Books, Park End Place, Oxford OX1 1HN
(Phone: 01865–241249; Fax: 01865–794449)
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www.oxbowbooks.com
Cover image: Ursus americanus baculum, mandible and innominate,
photo taken by Haskel Greenfield
Printed in Great Britain at
Antony Rowe, Chippenham

Contents
Preface .............................................................................................................................................................................. vii
Umberto Albarella, Keith Dobney and Peter Rowley-Conwy
Acknowledgments ............................................................................................................................................................ ix
Introduction
1. Vertebrate Demography by Numbers: Age, Sex, and Zooarchaeological Practice ..................................................... 1
T. P. O’Connor
Part I. New Approaches to Ageing and Sexing
2. Using Osteohistology for Ageing and Sexing ............................................................................................................. 9
K. Dammers
3. A Method to Estimate the Ages at Death of Red Deer (Cervus elaphus) and Roe Deer (Capreolus capreolus)
from Developing Mandibular Dentition and its Application to Mesolithic NW Europe .......................................... 40
Richard J. Carter
4. The Table Test: a Simple Technique for Sexing Canid Humeri .................................................................................. 62
Deborah Ruscillo
5. Sexing Fragmentary Ungulate Acetabulae ................................................................................................................ 68
Haskel Greenfield
Part II. Testing Existing Methods of Ageing
6. Reconciling Rates of Long Bone Fusion and Tooth Eruption and Wear in Sheep (Ovis) and Goat (Capra) ......... 87
M. A. Zeder
7. Accuracy of Age Determinations from Tooth Crown Heights:
a Test Using an Expanded Sample of Known Age Red Deer (Cervus Elaphus) .................................................... 119
T. E. Steele
8. Methodological Problems and Biases in Age Determinations: a View from the Magdalenian .............................. 129
J. G. Enloe and E. Turner
9. A Bayesian Approach to Ageing Sheep/Goats from Toothwear ............................................................................ 145
A. R. Millard
Part III. Studies in Ageing by Dental Eruption and Attrition
10. Tooth Eruption and Wear Observed in Live Sheep from Butser Hill, the Cotswold Farm Park
and Five Farms in the Pentland Hills, UK ................................................................................................................ 155
G. G. Jones
11. Determining the Age of Death of Proboscids and Rhinocerotids from Dental Attrition ........................................ 179
S. Louguet
12. Tooth Wear in Wild Boar (Sus Scrofa) .................................................................................................................... 189
O. Magnell

vi Contents
Part IV. Applications of Osteometrics and Epiphyseal Fusion
13. Phenotype and Age in Protohistoric Horses: a Comparison Between Avar and Early Hungarian Crania ............. 204
L. Bartosiewicz
14. Documenting the Channel Catfish Population Exploited by the Prehistoric Inhabitants
of the Station-3-avant Site at Pointe-du-Buisson, Southern Québec (Canada) ...................................................... 216
M.-E. Brodeur
15. Epiphyseal Fusion in the Postcranial Skeleton as an Indicator of Age at Death of European Fallow Deer
(Dama dama dama, Linnaeus, 1758) ........................................................................................................................ 227
R. F. Carden and T. J. Hayden
16. Size Variability in Roman Period Horses from Hungary ........................................................................................... 237
K. Lyublyanovics
Part V. Studies in Sexual Dimorphism
17.Environment, Body Size and Sexual Dimorphism in Late Glacial Reindeer ............................................................. 247
J. Weinstock
18. Sexual Dimorphism in the Postcranial Skeleton of European Fossil Elephants ...................................................... 254
L. Sedláčková

vii
Preface
Umberto Albarella, Keith Dobney and Peter Rowley-Conwy
This book is one of several volumes which form the pub-
lished proceedings of the 9th meeting of the International
Council of Archaeozoology (ICAZ), which was held in
Durham (UK) 23rd–28th August 2002. ICAZ was founded
in the early ‘70s and has ever since acted as the main
international organisation for the study of animal remains
from archaeological sites. The main international confer-
ences are held every four years, and the Durham meeting –
the largest ever – follows those in Hungary, the
Netherlands, Poland, England (London), France, USA,
Germany and Canada. The next meeting will be held in
Mexico in 2006. The Durham conference – which was
attended by about 500 delegates from 46 countries – was
organised in 23 thematic sessions, which attracted, in
addition to zooarchaeologists, scholars from related dis-
ciplines such as palaeoanthropology, archaeobotany, bone
chemistry, genetics, mainstream archaeology etc.
The publication structure reflects that of the conference,
each volume dealing with a different topic, be it
methodological, ecological, palaeoeconomic, sociological,
historical or anthropological (or a combination of these).
This organisation by theme rather than by chronology or
region, was chosen for two main reasons. The first is that
we wanted to take the opportunity presented by such a
large gathering of researchers from across the world to
encourage international communication, and we thought
that this could more easily be achieved through themes
with world-wide relevance. The second is that we thought
that, by tackling broad questions, zooarchaeologists would
be more inclined to take a holistic approach and integrate
their information with other sources of evidence. This also
had the potential of attracting other specialists who shared
an interest in that particular topic. We believe that our
choice turned out to be correct for the conference, and
helped substantially towards its success. For the pub-
lication there is the added benefit of having a series of
volumes that will be of interest far beyond the restricted
circle of specialists on faunal remains. Readers from many
different backgrounds, ranging from history to zoology,
will certainly be interested in many of the 14 volumes that
will be published.
Due to the large number of sessions it would have been
impractical to publish each as a separate volume, so some
that had a common theme have been combined. Far from
losing their main thematic focus, these volumes have the
potential to attract a particularly wide and diverse reader-
ship. Because of these combinations (and because two
other sessions will be published outside this series) it was
therefore possible to reduce the original 24 sessions to 14
volumes. Publication of such a series is a remarkable
undertaking, and we are very grateful to David Brown and
Oxbow Books for agreeing to produce the volumes.
We would also like to take this opportunity to thank the
University of Durham and the ICAZ Executive Committee
for their support during the preparation of the conference,
and all session organisers – now book editors – for all
their hard work. Some of the conference administrative
costs were covered by a generous grant provided by the
British Academy. Further financial help came from the
following sources: English Heritage, Rijksdienst voor het
Oudheidkundig Bodemonderzoek (ROB), County Durham
Development Office, University College Durham, Palaeo-
ecology Research Services, Northern Archaeological
Associates, Archaeological Services University of Durham
(ASUD), and NYS Corporate Travel. Finally we are ex-
tremely grateful for the continued support of the Wellcome
Trust and Arts and Humanities Research Board (AHRB)
who, through their provision of Research Fellowships for
Keith Dobney and Umberto Albarella, enabled us to under-
take such a challenge.
The present volume publishes the proceedings of the
session ‘Ageing and Sexing’, which was among the first to
be proposed for ICAZ 2002 and ended up being one of the
strongest and best attended. This was in large part due to
Deborah Ruscillo’s excellent organisational skills, but also
to the inherent interest and appropriateness of this subject
for an ICAZ conference. Whether we study material from
Argentina or Japan, from the Palaeolithic or the medieval

viii
period, we still need to deal with the issue of ageing and
sexing animal bones. A methodological session may be of
little interest outside the field of zooarchaeology, but this
is compensated for by the fact that all animal bone special-
ists will be interested in it. Initially Deborah wanted simply
to find the best venue to present her interesting new
method of sexing mammal bones through shape analysis.
However, here was an opportunity to be more ambitious
and organise a whole session dedicated to sexing and
ageing studies. Things went ahead as planned and this
book represents the culmination of almost three years of
work, begun with a cosy conversation in the warm environ-
ment of the Ruscillo/Cosmopoulos home in Winnipeg (as
the external temperature approached minus 30°C!).
The publication in 1982 of the volume “Ageing and
sexing animal bones from archaeological sites”, edited by
Bob Wilson, Caroline Grigson and Sebastian Payne, repre-
sented a milestone in the development of zooarchaeological
studies, and the book is, unsurprisingly, one of the most
cited publications in zooarchaeology. Since then, as Terry
O’Connor highlights in his introduction to the present
volume, much more work has been done in refining ageing
and sexing methods and in improving our understanding
of body development and sexual variation in the vertebrate
skeleton. However, so far zooarchaeologists are still by
and large adopting ageing and sexing methods that are
pre- rather than post- 1982. The challenge of this book is
therefore not just to add more information, but also to
persuade zooarchaeologists that the time is ripe for ex-
perimenting with new methods and for analysing data by
taking into account the substantial new advances that this
discipline has produced in more recent years. Only time
will tell if this volume will have achieved this ambitious
goal, but whatever the case, we have little doubt that it will
represent an indispensable tool for zooarchaeologists
worldwide.
Final special thanks must go to Vasili and Marilena
Cosmopoulos (Deborah’s son and daughter), who had the
good grace to be born during the final stages of the editing
of this volume. We could not have expected a better omen
for the success of the book.
Preface

ix
Acknowledgments
These proceedings are a direct result of the teamwork and
collegiality of the authors involved. Always polite and
accommodating, the participants of the Ageing and Sexing
Session were a pleasure to work with, and I thank them for
their cooperation and their contributions to methodology
in zooarchaeology. The participants of the session also
acted as referees of the published proceedings; each
participant reviewed two papers submitted for publication
to ensure the quality and accuracy of the information
presented herein. For the sake of keeping costs low, raw
data for the various studies presented in this volume could
not be published. The authors are happy to provide raw
data from their research upon request (addresses provided
at the end of each chapter).
The session would not have been such a success were
it not for the tireless support and direction of the confer-
ence organizers. This publication was made possible by
these same individuals who also bravely took on the series
editing and organization after the conference ended. On
behalf of all the participants of the Ageing and Sexing
Session, I wish to express our appreciation for the commit-
ment of Umberto Albarella, Keith Dobney, Peter Rowley-
Conwy and Deborah Jaques for the conference prep-
arations and publication series organization. I would also
like to thank Simon Davis and Caroline Grigson for chairing
the morning and afternoon sections of the ageing and
sexing colloquium, and also for acting as referees for some
papers submitted here.
Travel and accommodation grants for the participants
were supplied thanks to generous funding from the
Institute for Aegean Prehistory (INSTAP). We are grateful
for their support and their broad vision of archaeological
research. INSTAP is one of the few organizations that
realize the potential of zooarchaeological studies in the
quest of studying ancient peoples.

9Using Osteohistology for Ageing and Sexing
2. Using Osteohistology for Ageing and Sexing
K. Dammers
9th ICAZ Conference, Durham 2002
Recent Advances in Ageing and Sexing Animal Bones (ed. Deborah Ruscillo) pp. 9–39
Introduction
Over three hundred years ago, Clopton Havers (1691)
gave the first descriptions of the fine structure of bones
(Fig.1). The microscopic analysis of bone structure now
provides actual and potential methods of determining
the age-at-death and sex of vertebrates. Depending on
such factors as the availability of bones and their con-
dition as well as the age and species, histological analysis
can be more certain and more accurate than other
methods. In addition, it can also be used on material that
can not be applied to other methods.
This chapter begins with a summary of the relevant
physiology, focusing on osteons and growth rings; ex-
plains the principles of sample preparation; gives a survey
of histology for determining age-at-death, including
methods and results from dental and bone growth rings
as well as from osteons; and finally, presents a perspective
on using osteons for sexing. The chapter ends with a
glossary explaining common terms from osteohistology.
Relevant classic and recent literature is presented in the
bibliography.
Microscopic analysis of bones, teeth, and other animal hard tissue can yield considerable information. The most
intensively studied of such structures are growth rings and osteons. “Growth rings” accrue to the periosteal surface
of many bones as well as in teeth, reflecting annual and shorter periods. Where bone rebuilding has not destroyed
them, they can often provide an accurate ageing device despite some problems with readability. The mandible is
generally the best bone for this kind of ageing. Various vertebrates have been studied, with differing degrees of
accuracy. “Osteons” refers to systems of conduits (and associated structures) for small blood vessels and nerves
that grow and run lengthwise in much compact bone of many vertebrates. Osteons respond to the stress and strain
of life, and this allows for their use in ageing bones. Various bones, e.g. femur, humerus, and rib, have been used
with differing degrees of success and accuracy. Two general approaches are used: quantitative and qualitative. The
latter, used in continental Europe, appears to be easier, more objective, and probably more accurate, but both
consist of determining the relative frequency or area of various kinds of osteons, their parts and/or their fragments.
Much less effective and concerted have been attempts to use osteons for sexing, though under certain conditions this
can be done. The methods and techniques commonly used for studying osteons, including the preparation of thin-
sections with a microtome or taking of microradiographs, are not very difficult, but they do take some time to learn
and to carry out. Advantages of using osteons include their being relatively resistant to fire and other abuses
common to archaeological settings, their being able to be studied in small fragments of bones, and their presence
in robust bones. Most but not all osteon work dealing with ageing and sexing has been done on human materials.
A definite need exists for extending these efforts to more taxa, especially other long-lived ones and ones with marked
sexual dimorphism.

10 K. Dammers
What Is Osteohistology?
Osteons and osteohistology – the microscopic study of
bone – in general, constitute under-utilized sources of
information from the fossil record. They can be used in
determining taxa, nutrition and diet, stress, limb use,
sex, pregnancy, parturition / lactation, illness and injury,
age-at-death, and certain environmental factors. This
paper presents an overview of actual and potential uses
of osteons and other osteohistology for ageing and sexing
animals. Of necessity, there are certain generalizations
and simplifications.
What Are Osteons?
Bone is not static: it keeps growing, remodelling, breaking
up and dying through-out the life of the individual.
Furthermore, bone is in constant physical and chemical
contact with other parts of the body, helping maintain its
homeostasis. The life-lines of much compact bone are
called Haversian canals, the core of so-called osteons
(Haversian systems). These canals, about 20–300 µm in
diameter (depending on age and species) contain blood,
lymph, and nerve vessels, and meander along the long
axis of bones. The Haversian canals also send out smaller
lateral vessels, known as Volkmann’s canals, to provide
nutrition to the nether regions of the bone (Figs 2, 3).
One can classify osteons in various ways: for convenience,
we can think of primary (at the beginning), secondary
(or replacement or remodelled), and fragmentary osteons.
Fig. 1. Cover page of Clopton Havers’s pioneering work on the microscopic structure of bone. (From a copy at the
Göttingen State and University Library, Germany).
As the mandible, the shafts of long-bones, and other
bones grow, new osteons burrow through, insuring the
effective transport of nutrients (Fig. 2). The canals are
formed by osteoclasts (destructive cells) boring through
the established bone by “resorbing” it, affecting primary
bone, primary osteons, or other secondary
osteons, making a cone of construction in front of the
vessel. Osteoblasts (constructive cells) then line this cone
and secrete mineral-building substances, turning it into
a tube of minerals. This lining is said to consist of a
lamella, 3–20 µm thick made up of collagen fibres, ca.
0.06 to 0.6 microns in diameter. Over time, other con-
centric lamellae form inside the first, the outer edge of
which has a distinctive cement ring. Thus, a cross-section
of a secondary osteon consists of blood, lymph, and nerve
vessels, and perhaps up to seven to 20 lamellae
surrounding the canal. Within the lamellae are lacunae:
generally oval-shaped spaces with osteocytes trapped in
them but connected to each other with a fine web of
canaliculi containing their connective processes and the
central canal but not extending past the cement line.
The presence, frequency, size, shape, etc. of osteons
and their constituents will vary according to species, type
of bone, and location along the bone shaft. Other factors
affecting osteons are whether they occur near the core or
exterior of the bone, as well as age, season, sex, stress,
diet, injury, disease, and taphonomy (for archaeological
bone).

11Using Osteohistology for Ageing and Sexing
Change in Osteons
As the body continues to grow and age, various physical
and chemical stresses and strains play upon the compacta
and its component osteons. The lamellae, forming con-
centric rings around the Haversian canals, gradually close
them off, like a calcified water pipe. Replacement
(secondary) osteons cut through the old ones, leaving
osteon fragments, like the wreckage of old drainage pipes
beneath the streets of a city. The amount and area of both
the replacement osteons and fragments, as well as
resorption lacunae, generally increase with age. Not
surprisingly, different bones and bone locations, in
populations and sub-populations (such as sex) have
different stress factors, which are reflected in osteon
assemblages. These are “biographies”: the moving finger
having written, moves on but it leaves behind its message.
Not all mature compact bone is characterized by
osteons. Some bones in animals have different structures,
moderately correlated with gross taxon (e.g. horse or
reptile) as well as bone function and rate of development.
Thus, plexiform bone is formed in rapidly growing long
bones that need to endure heavy stress. Oddly, this
structure does not necessarily entirely preclude the
presence of an odd osteon or, following Wolff’s Law of
stress response, of clusters of them in strategic locations
(at least as time passes). In addition, there can be layers
of lamellae that can be added to the inner and outer bone
surfaces; these layers are discussed below, in the section
on ageing.
I also want to consider an even finer scale for a brief
moment before turning to lab techniques and ageing and
sexing. The molecular level of bone, in particular osteons,
is active. It is also not completely understood and has
suffered, as has histology in general, from a seldom-
broken focus on a limited number of species (especially
rats and humans). In any case, the standard model holds
that within a few weeks after initiation, e.g. due to an
injury or accumulated microfractures, a completely new
secondary osteon will be formed by the construction crew
(the BMU: bone remodelling unit). This unit consists of
the osteoclasts with acidic mineral-dissolving secretions
and is followed by the osteoblasts laid down along the
walls of this advancing cone. Some of the osteoblasts get
trapped and become or get designated as osteocytes. The
nutrient supply lines, with blood, nerve, and lymph
vessels, follow. The osteocytes are attached to the vessels
in the heart of the canal via extremely small vessels called
canaliculi, and thereby supply the bone with its necessary
life-giving provisions. After a while, a new lamella gets
laid down by osteoblasts, with lacunae occupied by further
osteocytes (old osteoblasts). In some places, lateral sub-
canals branch out, never into any other living secondary
osteon, but sometimes to the bone surface. And so it
goes, until the concentric lamellae close off the canal or
a new demand calls for rebuilding and the BMU cuts
through the established osteon. In humans, a new osteon
can lay down 70% of its mineral structure in a fortnight
or so, and be completed in 100 days, while completion in
a cat takes 50–90 days (Pritchard 1972). While turn-over
is said to be about ten percent per year, the natural life-
span of a secondary osteon is variously said to be about a
decade and a half to a quarter century. For a more detailed
but still archaeologically orientated discussion of osteon
histology, see Robling and Stout (2000), especially pages
187–190. Other detailed descriptions, including those of
the molecular and chemical processes involved, can be
found in bone physiology texts such as Bloom and Fawcett
(1975), Caplan (1998), Martin et al. (1998), Kingsley
(2001) and Currey (2002), or the multi-volume reference
work edited by Bourne (1972).
As indicated above, in some dense bone, Haversian
systems are absent or rare. Here, the nutrients are supplied
differently. The most prominent dense bone that is non-
osteonal is plexiform bone (though there can be some,
usually scattered, osteons in this kind of bone; see Fig.
4). Plexiform bone is so called because of the vascular
plexuses (an interwoven combination of elements in a
Fig. 2. Schematic of osteon. A. Cross section. Indicated
are Haversian canal with blood and lymphatic vessels,
a nerve, and loose connective tissue (a), osteocyte (b),
canaliculus (c), lamella (d), and reversal line (e).
B. Formation shown in longitudinal section (line of
advance is downward). Indicated are osteoclast dissolving
old bone (1), cutting cone (2), osteoblasts being deposited
(3), osteocyte (4), reversal line (5), closing cone (6), and
Haversian canal. (Adapted from Kardong 1995, fig. 5.26,
Robling and Stout 2000, fig. 7, and Walker and Liem
2000, fig. 6.9).

12 K. Dammers
cohering structure) contained within lamellar bone be-
tween non-lamellar bone (Martin et al. 1998). It has
been described as “bone in which the lamellae are grouped
into layers separated by well-developed vascular channels
or by bands of osteons” (Sheffield, no date). In cross-
section, it generally looks like corrugated cardboard that
has been fan-folded. This structure is observed in many
bones exhibiting rapid growth, e.g. equid long bones.
Plexiform structure can occur at certain locations, e.g.
the diaphysis of a bone. Some birds have what could
perhaps best be called plexiform structure, though it is a
bit aberrant. The structure is generally absent from most
living reptiles.
Other dense bone can be acellular or even non-vascular,
so that much of many bones of specific vertebrates (e.g.
marsupials and insectivores) is non-Haversian.
What Are Growth Rings?
On many animals (and practically all mammals if not
vertebrates), the hard structure adds periodic layers to its
surface, giving an appearance similar to the familiar rings
in trees (Fig. 3). These lines/layers/rings are similar to
tree rings in that they reflect, to a certain extent and
insofar as they are not obliterated by remodelling, 1)
periodicity (useful for ageing), and 2) stress (useful for
ageing and sexing). Among the hard tissues which exhibit
these predominantly microscopic accretions are long-
bones, ribs, and mandibles of a number of mammals and
reptiles. Turtle scutes, mollusc shells, otoliths, scales,
tusks, teeth (dentine, enamel, and cementum) and antlers
also exhibit periodic layers. I will go into the causes and
meaning of these so-called recording structures or growth
rings in the section on ageing for bones and, briefly, for
teeth. These lines are caused by (changes in) stress,
feeding, parturition, nursing/lactation, and other
behavioural cycles, internal biochemical triggers, and by
external cycles. Stress comes in many forms, some of the
important ones being the transition from intrauterine life
to that in the external world, the onset of puberty, sexual
activity, midlife change, and death. In addition, diet,
exercise, injury, disease, and seasonal weather vary the
stress on animals in ways that can be reflected in the fine
structure of the hard tissue.
Generally, incremental growth markers can be can be
divided into two types: bands and lines. The broad bands
represent rapid growth, whereas the thin lines reflect
slow growth, e.g. during hibernation or between feeding
periods, when deposits build up almost on top of each
other. In a sense, feeding is a source of stress. In any
case, daily feeding/rest cycles are reflected in dental lines,
while longer cycles (especially in some hibernating
animals) are more dramatically seen in hard-tissue
growth-ring patterns. Hard winters (and/or the dietary
difficulties caused by them) provide us with winter growth
ring patterning in cervids and some other temperate, sub-
arctic, and arctic animals, while wet/dry seasonal patterns
are reflected in the bones of lower latitude grazers and
reptiles. But even in less fluctuating climates, there
appears to be an annual pattern in at least some animals
visible to researchers, e.g. weekly and sub-hibernating
cycles.
The suprachiasmatic nucleus (SCN) (Ohtsuka-Isoya
et al. 2001), a part of the brain that regulates various
circadian rhythms, has been demonstrated to play a
crucial role in growth ring production. These various
factors interact, e.g. sickness and health, as the organism
varies its diet, sleep, sex, and other patterns in order to
protect the body and live. While this interaction makes
for more difficult interpretation, since the various “texts”
(osteons, bone growth rings, and various dental growth
rings) are “written” by differing combinations, a
composite analysis can theoretically reveal many a sub-
text. Thus, while this chapter deals with age and sex, the
investigator needs to keep in mind the environment,
health, etc. of the individual while doing an analysis.
With growth rings, there can be a certain amount of
error in the reading. In part this has to do with the
experience and lab tradition of the researcher, as well as
the equipment, but other factors play a role as well.
Important among these are the variability of growth rings
(disappearing, single, splitting, double, [Klevezal 1996,
61–70; O’Brien 2001, 50]), the differences from bone to
bone and from locus to locus on a bone and even around
a single cross-section. For each category and for each
species, and even macro-environment, each lab needs to
set up a reference series so that other workers can make
comparisons. For example, standards from the useful
work on mammals in Russia carried out by Galina
Fig. 3. Cross-section schematic of long-bone micro-
structure. Indicated are (A) line of arrested growth, (B)
primary osteon, (C) line of resorption, (D) Volkmann’s
canal, (E) resorption lacuna, (F) interstitial lamellae,
(G) secondary osteon, and (H) trabecular bone.

13Using Osteohistology for Ageing and Sexing
Klevezal can be compared and applied to Canadian
material. Still, researchers are already at the point where
for many taxa (most mammals found in Russia,
crocodiles, and many amphibians and cetaceans), major
bone growth rings can be taken as effective annual or bi-
annual markers except in long-lived species. In these
cases, bone turn-over destroys at least some rings, but
dental annuli are effective on the daily and annual level
and have even been reported to have weekly and monthly
variants (Klevezal 1996, 70–80). The preferred location
for bone growth-ring investigations is on the mandible
just below the ramus, while molars or canines cross-
sectioned near the gum line are the best selections for
most teeth. Osteon changes are best seen mid-shaft on
the tibia, femur or humerus, with the ribs also having
some value. Mandibular readings have not proven to be
very good in larger mammals, but in smaller animals
they hold out some promise of perhaps being equal or
superior to long bones for supplying useful records.
Sample Preparation
Introduction
The two most common research uses of fossil or archaeo-
logical osteons and other microstructures are taxon
determination (e.g. Máytás 1927; Amprino and Godina
1947; Enlow and Brown 1956–58), and ageing (e.g.
Yoshino et al. 1994, Klevezal 1996, Robling and Stout
2000). Once the bone and taxon have been determined,
samples can be examined for ageing (and/or other fields
of interest such as sex, disease, injury, and environmental
influences/reflection). While what follows is directed
specifically at the examination of bone for osteons and
growth rings, at least the general principles are applicable
to the histological study of other hard tissues.
The procedures for preparing samples for study are
similar, independent of sample or research question, as
long as one is using thin sections and a standard micro-
scope. Variations occur primarily with the stability of the
available bone and which observation methods are to be
used. This is especially the case in choosing between
“blind” approaches that are relatively rigid (in order to
be consistent and objective) and “choice” approaches
which require the researcher to select an appropriate field
of view.
Sampling
If a population is being sampled, two prime consider-
ations are to be observed. First, the sampled material
must be of adequate quality. Unfortunately, this cannot
always be determined before processing, though general
appearance is a rough guide and can be greatly assisted
by a knowledge of the local taphonomic conditions. The
second consideration is to follow usual statistical and
sampling procedures.
If an individual is being sampled, an appropriate bone
should be selected. In most species, the bones found to
give the best results are the femur, humerus, tibia, and to
a lesser extent the ribs, for osteons. For growth rings,
extensive successful research on the mandible makes it
the current bone of choice. While taking numerous
samples from a single bone has been legitimately en-
couraged (e.g. Harsányi 1993), it seems that this is not
normally necessary (Robling and Stout 2000; Klevezal
1996). Instead, one location at about midshaft should
generally suffice. The sampling technique will determine
whether a disk or more restricted shape (plug or shaft)
should be taken from the bone. Limited disposability of
a bone also restricts the shape and amount of the sample
taken. If possible, a disk-shaped sample is preferable
because it provides all possible loci and rings and is thus
applicable to all analytical techniques. The section should
be taken perpendicular to the long axis of the bone.
Physical Preparation
Once the area to be sampled has been chosen, the slice is
removed from the bone with a fine hacksaw or band saw.
The slice should be about a centimeter or a half a
centimeter thick. It is then embedded in resin (balsam) to
keep it stable, whether it is fossilized or not. After a full
day to harden, the block is then put on a special cutting
machine called a microtome and cut to the appropriate
thickness (ca. 30–80 microns), creating a smooth surface.
The slicing takes a few minutes, depending on the size
and hardness of the bone. The sample is now ready for
mounting on a slide, depending on the observation
method. This is the method used in most better-equipped
labs. It requires very little skill; however, microtomes
are costly. An alternative approach presented by Maat et
al. (2001), requiring no expensive equipment and
producing better quality samples, requires somewhat more
skill and practice. Basically, in twenty minutes a hand-
ground sample of the highest quality will be produced.
Other guides to preparing the slides include Chinsamy
and Raath (1992), Wilson (1994), and Pfeiffer (2000,
289–90). For dental samples, see Beasley et al. (1992).
The most frequently used method to examine the
samples for growth rings or osteonal structures involves
an optical microscope with transmitted light and an
adjustable polarizing (birefringent) filter, at magnifi-
cations from about 30× to around 200×, depending on
species, etc. In this case, the sample is simply mounted
untreated on a slide, usually without any hues, though a
few researchers use various stains, such as light red Gram-
Weigert, toluidine blue or basic fuchsin. Teeth, especially
if demineralized, are normally stained, with haematoxlin
or toluidine-blue (see Klevezal 1996, 5–13). On the other
hand, O’Brien (2001, 47–8; see also Lieberman and
Meadow 1992, 59–69 and Stutz 2002) argues that teeth
studies should use the following standard procedures in
order to get results that truly reflect annual growth rings:

14 K. Dammers
undecalcinated teeth should be embedded in a fixative
and petrographically thin-sectioned without staining
before being examined. A transmitted-light microscope
using cross-polarized light should be used to examine
the acellular cementum. The increments are then defined
on the basis of refractive properties. In fact, the alternating
lines are not always clearly distinguishable, at least for
the novice even with the cross-polarized light, though
O’Brien (2001) claims a consistency and accuracy of
readings.
Alternative viewing possibilities that have been used
are reflective light microscopy, SEM, microradiography,
and scanning acoustic microscopy (Katz and Meunier
1993; Brandt and Klemenz 1998; Eckardt and Hein 1999;
Katz and Bumrerraj 1999; Smitmans et al. 2000). For
SEM, acid etching of non-fossil material can improve
differentiation. Finally, some researchers have begun
producing three-dimensional images of osteons by
computer image manipulation of stacked slices (Stout et
al. 1999), direct-reading of sliced-off surfaces (Constans
2001), CT scans (Pattijn et al. 1999; Fajardo et al. 2002)
with a resolution of 60 microns, and non-destructive X-
rays down to two micron detectability (Tappen 2001;
Skyscan, no date). While these advanced techniques
provide a more realistic and complete picture of bone
histology – at least at the osteon level – they have yet to
be incorporated into the interpretive body of knowledge
based on our “flat-earth” model. Still, they bode a
revolution, even though they are not (yet) of the high
resolution that cross-sections are and are not capable of
presenting all relevant details. Even finer studies,
focusing on third order, or ultrastructure, such as atomic
force microscopy (Henning et al. 1999), are just now
being applied, as is Raman spectroscopic imaging (Timlin
et al. 2000).
Once the section has been fixed for a microscope slide,
it can be examined with any number of devices: reflected
or transmitted light, SEM, SAM, STM, AFM, etc. As
stated above, though, the most common approach is to
use transmitted light with a variable polarizing lens.
Because using polarized light takes a certain amount of
fidgeting and since different viewings can provide dif-
ferent information, most investigators prefer to work with
the original material, although a minority prefers the
objectivity of a fixed photographic image. For qualitative
and growth ring work, a low resolution (perhaps 10×–
20×) is first used to scan for an area that has not been
disturbed by anomalies, such as those caused by injury,
disease, muscular stress, taphonomy, or the lab dog. When
a good area has been determined, then a magnification of
about 50× to 100× (or more for growth rings in some
taxa, and for tooth enamel and dentine 70× to >250×) is
selected and the polarization can be tried out. The slides
should of course be clearly and systematically labelled to
include all relevant information concerning the source: a
histological sample without adequate documentation is
about as useful as a sherd or flint chip without
provenience. If feasible, remaining bone or a significant
portion of it should be retained with the sample. This is
especially important if one chooses to work with images
rather than physical samples.
Determination of Age-at-Death
Introduction
The living world is ruled by three clocks: that of the sun,
of the moon, and of the earth. Most clearly laid down in
animal hard tissue are the marks of the sun, that is, the
annual record. Next come those of the earth. The smallest
astral body, the moon, lays down the most controversial
and some of the most precise of these marks: menstrual
and estrual stress markers on the one hand and tidal
markers on the other. Yes, even tidal markers with ages
reflect changes as fine as a half day or so. Furthermore,
the moon in its four phases (or the 6.6 day cycle of solar
radiation) could be related to the weekly records that
some researchers have reported! Besides these forces,
which leave their records as incremental marks, there is
the less precise but more robust Haversian system, which
can be used as well, especially when the growth rings are
defective, missing or ambiguous. It can also be used as a
check on the growth-ring records.
Fig. 4. Cross section of sheep’s long bone showing
primarily plexiform structure with some primary osteons,
especially at bottom of picture. (From Demeter and Máytàs
1928, fig. 85; © Springer-Verlag).

15Using Osteohistology for Ageing and Sexing
Growth Rings
Various researchers have demonstrated that the hard
tissue of molluscs and practically all vertebrates have
annuli, rings or lines that reflect annual and sometimes
even finer cycles. Annuli can be found in fish bones and
scales (Menon 1950; Das 1994), amphibian bones (Enlow
and Brown 1958), reptile bones (Horner et al. 1999),
tusks (Fox 2000), enamel, dentine, cementum and mam-
mal bones (Klevezal and Kleinenberg 1969). In these
rings are recorded the individuals’ responses to weather
(hot/cold, wet/dry) and the effects of hibernation, as well
as dietary, sexual and monthly stresses, mediated through
an internal clock that seems to have incorporated at least
days (ca. 25 hours) and years. The general pattern consists
of a growth ring (or recording structure) consisting of
both a growth zone (in which rapid growth produces a
wide band), and an adjacent narrower band representing
either no outward growth (line of arrested growth: LAG)
or markedly slowed growth (annulus). An annulus can
contain one or more LAGs. Ideally, there is one clearly
defined growth ring per time unit – usually a year. These
get added on centrifugally to the periosteal surface.
Some taxa can present various problems such as filled
pulps precluding ageing of older individuals, eroded
cement, lack of teeth, resorption of adhesion lines in
bone, or accessory and double lines. Despite these dif-
ficulties, Klevezal and Kleinenberg aver that “the method
for determining the age of mammals according to the
number of layers in the tissues of tooth and bone is the
most universal and accurate” (1969, 109). This pers-
pective seems correct to me and, mutatis mutandis,
applicable to most other hard-body animal taxa in polar,
temperate, rainy/dry, and, less auspiciously, in monotonic
climes.
Dental Growth Rings
Dental growth rings are usually the most detailed and, in
general, give the finest-tuned record of growing older.
Although teeth are not the focus of this article, the
following is a very brief introduction to these related
phenomena. For an overview, see Carlson (1990, 542–4)
or Pike-Tay et al. (1999, 297–301); Reid (1997a) for
dinosaur histology; for mammals, see Klevezal (1996,
passim); and for an archaeologist’s bibliography, see
Gordon (1984 and 1992).
Most mammalian teeth can be said to have three kinds
of hard components, each capable of providing recording
structures. Starting on the inside is dentine, a 35% organic
material that grows inward in curved waves, gradually
filling up the core of the tooth. The lines of dentine can
have various meanings, depending on location and other
factors. Enamel forms the visible part of teeth. It is very
hard, being 96% percent mineral (Klevezal 1996, 15–6).
The part of the tooth that attaches it to the jaw is fittingly
called ‘cementum,’ a 50% inorganic material. “The
physical and optical expressions of growth increments in
teeth are due to differing patterns of collagen fibre
organization and to cell content / degree of mineralization”
(Pike-Tay et al. 1999, 298) and crystal orientation. For an
explanation of the complicated microstructure and sample
preparation of teeth, see Hillson (1986, 107–75);
Lieberman and Meadow (1992, 59–69); and Klevezal
(1996, especially 16, 24–48 and 70–80).
Half a century ago, V. B. Scheffer (1950) began studies
on pinniped dental growth layers. Laws (1952 and 1953)
then used pupping season-related rings of alternating
columns and marble dentine to age elephant seals’ canine
teeth of up to twenty years to within one month. He also
did preliminary work on other mammals that other
people, especially Klevezal and her associates, have since
developed in detail, and extended studies to enamel and
cementum. Enamel is now the material most commonly
used for ageing mammals with growth rings. It is believed
that “mammalian dentine universally shows circadian
increments” (Ohtsuka-Isoya et al. 2001, 1364). Dentine
has been demonstrated to lay down observable increments
on an approximately 24-hour cycle in rabbits, rodents,
humans, pigs, and other mammals, influenced or
controlled by the brain’s suprachiamsmatic nucleus
(Ohtsuka-Isoya et al. 2001). Klevezal (2001) reports that
incisors in female mice have clear daily lines in the
dentine (see also Trunova 1999 and Klevezal). Fine lines
of alternating high and low transparency, apparently
reflecting differences in mineral content (Klevezal 1996,
38), are laid down starting before birth. Heavier de-
marcation reflects longer-termed periods and isolated
events such as circumnatal trauma. Unfortunately, the
dentine growth, which advances in an inwardly filling
manner at rates per day of 12–24 µ in voles and 1.9–3.9
µ for mice (Klevezal 1996, 12), can completely fill up the
tooth and then stop recording, sometimes as early as the
onset of maturity.
Finding tooth sections with lines actually recording
rhythms rather than frequent multitudinous individual
variable or non-recurring events is difficult (Klevezal
1996, 155). Thus, dentine has not been used much in
ageing terrestrial mammal teeth (see Table 5 in Klevezal
1996, 54–9; see also Hillson 1986, 223–9, for an archaeo-
logical perspective). Work using this technique does
continue, however, including successful daily-line
readings in mice incisors (Klevezal 2001), hibernation
markers in 17 species of rodents grouped into three
patterns (Trunova and Klevezal 1999), and Russell’s
advocacy for its use in humans (1996). Additionally, Fox
(2000) claims to have identified yearly, weekly, and daily
incremental growth lines in gomphoteria from the Mio-
cene. Other studies reported in the literature include
Miani and Miana (1972) on dog dentine as well as Okada
and Mimura (1941) and Rosenberg and Simmons (1980)
on rabbits. The latter produced easily identifiable images
of daily increments (Fig. 5). One other group of animals
where dentine has been used is bats, creatures that
otherwise are quite a difficult to age. Their dentine

16 K. Dammers
incremental lines are often observable, and represent a
conservative minimum age in years that is, unfortunately,
characteristically well below actual age (Batulevicius et
al. 2001; Batulevicius, pers. comm.).
The histology of enamel is visually clear (Klevezal
1996,15–6) and also seems to reflect an acid/neutral
alternation (Takagi et al. 1998). In principle, it is a good
recording medium; however in practice, enamel often
fails due to attrition, resulting in the loss of recording
structures. The daily bands are also complete at eruption
(Klevezal 1996, 14). Thus, its application is pretty much
restricted to young individuals, where some of the
permanent teeth are not yet complete. On the other hand,
where applicable, daily increments have been claimed to
be quite readable, even though some oral biologists deny
the very existence of incremental lines (for discussions
see Klevezal 1996, 15–6; Hillson 1986, 119–27; Ramirez
Rozzi 1998)!
As mentioned, cementum is by far the most popular
source for obtaining growth rings in ageing mammals
(for methods see Beasley et al. 1992 and Stutz 2002; for
surveys see Stallibrass 1982 and Klevezal 1996; for a
simple explanation and archaeological references, mostly
to seasonality, see Pike-Tay et al. 1999, 297–305 or
Lieberman and Meadow 1992). In fact, according to
Peabody (1961, 12), in 1860 Hittel noted cementum layers
in California grizzly bears and called for experiments in
mammals to confirm their presence. A century later,
Smirnov (1960) undertook such studies, examining some
carnivores and rodents, complementing work that was
being done on marine mammals at about the same time.
Of the histological areas, cementum usually gives the
most correct and reliable ages, though not particularly
the most precise. It is particularly advocated for use in
carnivores (King 1991, 31; Klevezal 1996, 106), and its
use is popular in ageing ungulates as well (Klevezal 1996,
58–9; Lieberman and Meadow 1992). For carnivores,
the canines are preferred, whereas in the ungulates
incisors and molars are used. On the other hand, about a
quarter of sheep ages were reported as being erroneous
by more than a year. The “crystalline orientation and/or
size is responsible for the layered appearance of
cementum” (Cool et al. 2002, 386), which can be read
accurately as reflecting years, though advancing age and
double lines broaden the estimate to about 5%. Klevezal
notes that the “pattern of growth layers in cementum is
usually simple…, [consisting of] a band of tissue and an
incremental line,” which looks pale in transmitted light
and dark in reflected light (1996, 39). Most of Klevezal’s
data on growth rings, both her own work and that of
others that she discusses, focuses on the effective ageing
using cementum in most groups of northern mammals
(see 1996, Table 5, pp. 54–59 and Chapter 6, pp. 124–
177, and more recently Klevezal 2002). O’Brien (2001),
in reviewing the literature on African cementum studies,
concluded that, where the standard procedure (presented
in the previous section) is used and annual increments
are taken as a combination of one opaque (dark) and one
translucent (light) line, good results are obtained for
zebras and other tropical mammals.
Specific studies include Klevezal’s previously
mentioned study of mice (2001), which also reported on
cementum. She noted that winter incremental lines in
the molars not only could be used as an indication of
over-wintering, but that the amount of growth on either
side could narrow the age estimate. White (1974) studied
roe deer using a 20× reflected-light microscope to
examine sections he had ground smooth with very fine
carborundum. He observed clear and regular repeating
pairs of opaque and translucent lines, the pairs matching
the number of years lived. Preparation and evaluation,
described in detail, took thirty minutes per tooth. In the
Fig. 5. Stained and decalcified rabbit incisor dentine showing daily increments. Each pair of dark and light bands,
indicated by the lines above the photomicrograph, is about 20µm in width. (From Rosenberg and Simmons 1980, 32,
fig.1A; © Springer-Verlag).

17Using Osteohistology for Ageing and Sexing
discussion section, the author notes that earlier studies
indicate that the teeth of roe deer associated with a
demonstrably milder winter and higher quality of food
do not show winter markers. Pike-Tay (1995) noted that
seasonal markers on caribou teeth were made difficult to
read by a number of factors, including specimen
preparation and variations in feeding (see also Miller
1974 and Pike-Tay 1999). McKinley and Burke (2001)
found that cementum on horse teeth could be read with
only minor, resolvable, inter-observer variability, and
produce accurate results. A study of wildcats proved
effective to closer than one year, though with the caveat
that 7% of the animals had an additional (but identifiable)
“kitten” line (Garcia-Perea and Baquero 1999). The first
real line develops at nine months. Not so confident were
the results of an extensive study of fishers that questioned
accuracy on individuals three years and older, noting
that similar difficulties for other fur-bearing animals had
been pointed out in the literature (Arthur et al. 1992).
Studies on various primates have given good results
for molar cementum. For example, Kay et al. (1984),
studying eight wild-living macaques of known ages,
concluded that around 90% of the ages could be correctly
made, making cementum measurements “the most
accurate method for aging skeletally adult primates [yet
to be] tested on animals of known age” (85). On the other
hand, long-lived species can be problematic. Condon et
al. (1986) report errors averaging six years using a
modern population of geographically restricted Homo
sapiens, using cementum annuli. This is hardly the “to
the day” accuracy reported by some independent
researchers, especially in other species (329). In addition,
Condon and his co-workers note that inaccuracy increases
with age in a number of cervids (moose, red deer, and
Virginia deer) (328). In his recent book, O’Connor notes
“an uneasy consensus emerging that cementum increment
counts are an acceptable estimate of age in some species,”
especially when used on individuals that have not reached
later adult ages (2000, 82).
Closing on dental growth rings, we note their general
effectiveness, with each kind having advantages (dentine:
fine scale, early start, protection from destruction; enamel:
effective in young and even prenatal studies; cementum:
continues for all or almost all of life). The limitations
must also be recognized. Key among these are: dentine
can fill the pulp cavity and thus stop recording; enamel
is restricted to ageing younger individuals, and outer
layers can also be lost by wear; cementum can be resorbed,
be covered with calcium build-up, sometimes has double
layers and other reading problems, and the winter line
appears to be absent in non-stressed conditions on some
species. Finally, each is subject to its own diseases which
mar recording. Even though there is a general retrench-
ment (Darius Batulevicius, pers. comm.) from some of
the euphoric claims that have been made in the past, to
a considerable extent this retrenchment has been brought
about by inconsistent and less than optimal testing
methods (McKinley and Burke 2001). Careful develop-
ment of controls should continue to bring us closer to
highly accurate ageing where good tooth samples are
available.
Bone Growth Rings
The bone building crew (BMU: bone maintenance unit)
has workers all over, including on the endosteal (inner)
and periosteal (outer) surfaces of the bone. On both of
these surfaces, osteoblasts, apparently receiving orders
from nearby monitoring osteocytes (Moskilde 2001, 152),
lay down organic material that later becomes mineralized
by deposition of hydroapatite crystals. Given the right
conditions, this apposition will appear in the form of
layers, analogous to tree rings or cementum growth rings.
These are called circumferential lamellae. This process
is very complicated, and its workings are still a some-
what controversial matter (see Klevezal 1996, 17–23).
The growth rings will normally form if, when, and where,
a slowing of growth allows a concentration of mineral-
ization and/or change in orientation of the crystals. When
growth is rapid, it is in fibro-lamellar form; when it
slows, pure lamellar structure can form. Not all bone will
have these bands. For example, load-bearing and fast-
growing bones and fast-growing species rarely exhibit
them (at least in the loci so far investigated), especially
since the bone tissue is not rarely plexiform. Although
apposition continues throughout life (Klevezal 1996, 18),
it slows coincidentally with slowing in skeletal growth.
At this point, growth rings are more likely to be visible,
although in long-lived species, rings late in life can be
absent or almost piled up on top of one another. Un-
fortunately for the annuli-bedazzled archaeologist, the
bone-building crew does not stop once the circumferential
lamellae are laid down. Just like city planners redirecting
municipal services to dig up newly laid street surfaces to
put down new water or gas lines, new service lines –
secondary osteons – remodel the bone, often including
the growth rings. But that is another matter discussed
below. Where the resting lines are intact, with a so-
called reversal line marking the limit of reconstruction,
they are taken (based on extensive research) to reflect
annual periods of relative rest in growth during the winter
or dry season. To what extent this periodicity is due to an
internal clock controlled by hormones from the SCN (for
example) or to the environment has yet to be fully
understood. Bone growth rhythms probably evolved in
synchronicity with the seasons, though the evidence seems
to support the usual flexible mix of nature and nurture.
Because of geometric conditions, the endosteal (medullar)
surface growth rings are more constrained and generally
less useful for ageing. In some taxa, finer than annual
increments have been reported, but in most species, extra
lines represent artifacts (in the sense of Störfaktoren).
“Periosteal bone tissue as a recording structure is, in
general, characterized by high sensitivity and small period
of record persistence. Because of that it is the least

18 K. Dammers
convenient” of annuli to use for ageing given variations
due to environmental change and injury (Klevezal 1996,
106). On the other hand, it is useful in vertebrates without
useable teeth-structure and in individuals with no
remaining or serviceable teeth.
The following is a short overview of some of the
findings of periosteal growth rings.
Fish. The use of otoliths for ageing and seasonally
identifying various fish is generally effective and well
known (e.g. Victor and Brothers 1982; Van Neer et al.,
2002a). The annuli can reflect daily cycles, perhaps of
feeding and/or light, so that otoliths can provide
extremely fine ageing – down to the day – and, maybe
even the time of day of the kill! There are, however,
problems of misreading due to false, hidden, and missing
annuli (Victor and Brothers 1982). Otoliths are also not
as commonly recovered from archaeological sites as
vertebrae. Fortunately, these bones also have growth rings
that can also be used to age fish, though not as well as
otoliths. For discussions of fish vertebrae and other non-
otolith bony materials for ageing and seasonality, see
Menon (1950) for an extensive though old survey, and
Van Neer (1993: clariid pectoral spines), Van Neer et al.
(1999: plaice) and Dubick (2000: eagle ray) for recent
specific and critical work.
Amphibians. Castanet (1975) and others have also found
acceptable growth rings for amphibians. Examples in-
clude golden-striped salamanders (Chioglossa
lusitanica), where annuli appear to be laid down yearly,
and are different after metamorphosis (Lima et al. 2001).
A northern species of salamander could be aged within
two years of actual age, with no resorption difficulties
seen in the older individuals (Smirina et al. 1994). In
various anurans, growth rings in digits as well as in the
femur were effectively used (Sullivan and Fernandez
1999; Erişmiş 2002). Fig. 6 gives an example of how
these growth marks look in a stained decalcified cross
section. A recent study of sixty-two Indian frogs
(Microhyla ornata) produced consistent, almost perfect,
results for many bones in both sexes and all ages, thus
showing that extreme seasonal temperature swings are
not necessary to produce meaningful growth rings in
frogs (Kumbar and Pancharatna 2001). On the other
hand, Smirina and Makarov (1987) emphasized the
effects of climate on resorption in the grass frog (Rana
temporaria). Esteban et al. (1996) also caution that 7%
of the 103 Spanish frogs they studied showed weak, un-
readable winter markings. Other amphibian studies, with
generally the same results, have been reported in
Schroeder and Basket (1968), Kleinenberg and Smirina
(1969), Smirina (1972), Castanet and Roche (1981),
Guyetant et al. (1984), Castanet and Smirina (1991),
and Smirina et al. (1986). Caution should be exercised,
however, as a number of these studies note that in some
species resorption of inner growth rings can lead to
deceptively low readings, especially for the untrained
observer (Kumbar and Pancharatna 2001).
Reptiles. There is extensive and sophisticated literature
on growth rings in dinosaurs (e.g. Nopsca and Heidsieck
1933; Reid 1990, 1997; Chinsamy 1993; Horner et al.
1999), which has helped advance interpretive histology.
Other reptiles have been reported in some detail as well
(e.g. Bryuzgin 1939; Warren 1963; Castanet and Smirina
1991; and, for a Permain genus, Ray and Chinsamy 2004).
Donald Enlow, the leading comparative osteohistologist
of the mid-century, gave an overview of bone growth
rings for ageing reptiles which is largely applicable to
other vertebrates. He came to the conclusion that, while
growth rings can provide extensive data on past
circumstances of growth and broad-band relative ageing,
they are apparently not useable for absolute ageing. The
confounding factors he listed are: 1) the problem of
resorption, 2) the presumed reflection of seasonality of
annuli inconsistent with tropical and temperate snakes
showing similar patterns, 3) only certain loci having
rings, even on the same elevation of a bone shaft, one
side frequently having more laminae (1969, 63–68).
While these factors preclude a simple application of a
one-to-one correlation of laminae with years or over-
Fig. 6. Cross section of femur of a water frog (Rana
bedriagae). Indicated are lines of arrested growth are (a-
c), growth zones (1–3), osteons (Ost.), resorption line (rl),
endosteal bone (eb), and the medullary cavity (Mc). (From
Erişmiş et al. 2002, figure 2; © Turkish Journal of
Zoology).

19Using Osteohistology for Ageing and Sexing
wintering, further studies have shown that a careful
understanding of individual species does provide fairly
accurate quantitative ageing in a number of cases
(Castanet and Smirina 1991).
Bone growth rings in crocodilians have been ex-
tensively studied (Buffrénil 1980; Buffrénil and Buffetaut
1981; Hutton 1986). Crocodiles show clear annual growth
rings, although those held in monotonic environments
and provided with an abundant diet exhibit decidedly
less marked seasonality. Still, the lines do not generally
disappear (Ferguson et al. 1982, but see Horner et al.
1999, 299).
As early as 1939, Bryuzgin concluded that “in snakes,
like in fishes, in winter, during the period of cessation of
nutrition, the bone grows scarcely if at all” (1939, 404).
He found lines of arrested growth on the os transversum
of grass snakes (Natrix spp.) to consistently, clearly, and
effectively reflect the known number of over-winterings
(see also Petter-Rousseau 1953). Examination of his
illustrations (see Fig. 7) indicates that a certain amount
of skill in interpretation is required. Snakes were also
studied by Peabody (1958 and 1961), Castanet (1974)
and others, all reporting generally good annual cycles of
growth layers. Tropical snakes’ patterns are similar to
those of temperate snakes, allowing their annuli to be
used for ageing.
Lizards have been studied extensively, often with large
samples; the simple structure and frequent absence of
remodelling make their bones easy to work with. Rock
lizards’ (Lacerta) femurs have bone layering that has
proven effective for ageing to within a year or two despite
resorption (Castanet and Roche 1981; Smirina 1974),
though resorption can confound results (Smirina 1974).
What is particularly interesting is that the layers are the
same for both parthenogenetic and bisexual Lacerta
(Arakelyan and Danielyan 2000). The effects of a mild
climate on layer formation have also been studied for
lizards. Patnaik and Behera (1981) found apparent annual
lines on the bones of Galotes versicolor, but other
researchers have pointed out that the lines are less distinct
than those in lizards in temperate climates (Castanet and
Gasc 1986; Castanet and Baez 1991). Other lizards and
snakes are also reported to have useable annuli (Castanet
1974, 1978, and 1994; El Mouden et al. 1997; Buffrénil
and Castanet 2000).
Turtles have bone as well as shell growth rings (Mattox
1936). However, resorption often partially destroys the
former, and they reflect size more directly than age.
Castanet and Cheylan (1979) provide a correction factor
using carapace weight/length, allowing this bone data to
serve as a supplement to scute-dating. All these conditions
make it of only occasional value to the zooarchaeologist.
While reporting his findings in dinosaurs, Horner et al.
stated that simple counting of lines of arrested growth
“now appears to be potentially unreliable” (1999, 302).
In general, then, reptiles do have growth rings, but a
simple reading is not possible, and ageing must be done
with caution, taking various factors into account, and
even then, in some cases, the results will not be as precise
as one would like (Griffin 1962; Suzuki 1963; Enlow
1969, 67–69; Horner et al. 1999).
Avifauna. While well-marked layering is sometimes seen
in bird bones (e.g. Chinsamy et al. 1994), it does not
seem to be very promising for age determination in
avifauna, especially in light of the considerable resorption
that occurs (Hodges 1974). For example, Van Neer et al.
report that their data on modern chickens “show clearly
that there is no relation between the number of endosteal
layers and the age of the individuals…” (2002b, 132),
verifying their generally negative survey of the usefulness
of growth rings for ageing birds. On the other hand,
while Lapeña and her colleagues reported poor results in
mandibular growth lines from partridges, they did get
apparently accurate results from martins’ mandibles
(Lapeña et al. 1993, cited in Van Neer et al. 2002b,
128).
Mammals. The following survey highlights mammals that
are of particular interest due to growth ring quality or
special archaeological significance. Most of this material
has been gathered or reviewed by the leading authority
on mammalian growth rings, G. A. Klevezal (1996).
Generally, the locus of choice is the mandible just below
the ramus or last molar, though this is not always the
case. Additionally, when this site is not available or
adequate, other locations on the mandible or on other
Fig. 7. Os transversum of a nine year old grass snake
(Natrix natrix) with incremental lines (From Bryuzgin
1939, 404, fig. 3).

20 K. Dammers
bones can sometimes be used. Long bones and ribs are
among these other potential bones. Whether vertebrae
can be used, as they have been to a certain extent with
fish, seems not to have been investigated.
Pinnipeds were among the first mammals for which
correlations between bone growth rings and age were
developed, and almost all marine mammals have been
demonstrated to have bone and/or teeth growth lines.
Over three decades ago, Klevezal and Kleinenberg noted
that all pinnipeds have readable growth rings but included
the caveat that since resorption was a serious problem,
many loci have to be sampled (1969, 95). On the other
hand, Klevezal has more recently (1996, 157) noted
contradictory results. In addition, it seems that after
twenty years in males and ten years in females, the
mandibular annuli readings can lose their otherwise good
readability. In cetacea, the odontoceti have good growth
rings, whereas no rings have been reported for the
mysticeti (Buffrénil 1982; Klevezal and Kleinenberg
1969, 85, 86).
Few insectivores are long-lived. Shrews and others
not living over two years, have readable growth rings
indicative of over-winterings (Klevezal and Kleinenberg
1969, 44). Even longer-lived insectivores, specifically,
moles and hedgehogs, have good mandibular growth rings
coinciding in number with dental annuli (Klevezal 1996,
124–126). Admittedly, these and most other vertebrate
species reported have been examined in only small
samples and from geographically restricted areas, but
the overall pattern has been repeatedly substantiated.
A number of studies on lagomorphs all produced good
or very good results of winter-annuli correlations. For
example, a study of 57 European rabbits found only two
mandibles that did not have the same number of lines
and years of age – and these two were actually off by only
one month (Henderson and Bowen 1979). The effective-
ness of mandible growth rings in ageing lagomorphs is
especially encouraging, since the nature of hare and rabbit
teeth practically precludes their use in ageing (Klevezal
and Kleinenberg 1969, 49; Klevezal 1996, 54, 145, 146,
149). Pika over-winterings get recorded in their annual
layers (Klevezal and Kleinenberg 1969, 49).
Rodents, with their peculiar physiologies, e.g. usually
ever-growing teeth and a highly efficient calcium supply
system, have to be examined for different types of patterns.
Marmot mandibles provide accurate growth rings up to 4
to 5 years, but then often stop. Long bones seem to be
even less effective than mandible growth-ring ageing.
Hamster and souslik growth rings are good up to 3–4
years. Gerbil mandibles are apparently even less reliable
than other rodents, one winter being clearly marked but
further marks apparently having no chronological
meaning. Squirrel radii bones provide better annuli than
mandibles or any other tested bones, but additional lines
are frequent. Two important and very common rodents
have had a number of studies done on them. Mice
mandibles have good growth rings, with winter resting
lines over 90% effective. On the other hand, rats are
controversial. While some studies have found no
correlation of growth rings and age, others have reported
fairly good results. Interestingly, Klevezal reports that
experienced readers of vole growth rings were able to
accurately age rat mandibles using the same method.
One very important problem distinguishing a single year
is identifying “the difference in the formation of annual
layers in animals of early and late litters in the same”
year due to porosity of bone (Klevezal and Kleinenberg
1969, 79) (rodents: Klevezal 1996, 130–150; 2002. mice:
Klevezal 2001).
Bats have very fine bones, with special functions.
Possible growth rings are inadequate in number,
apparently due to apposition stopping (Klevezal and
Kleinenberg 1969: 48). On the other hand, promising
results have been suggested in alveolar bone (Schowalter
et al. 1978 as reported in Klevezal 1996).
Ungulates often provide the bulk of non-human bone
at archaeological sites. Thus, it is unfortunate that due to
resorption, growth rings are absent from the bones that
have so far been examined (including the ubiquitous long
bones (Klevezal 1996, 171–177). Compensating for this
absence are the effective lines in cementum for some
ungulates (see above), and the potential of osteon ageing
(see below).
At least smaller carnivores generally exhibit lines of
arrested growth in a number of bones, though the extent
to which these lines reflect annual cycles is not totally
clear, apparently under-reporting years (Klevezal 1996,
96; see also Pascal and Castanet 1978). Klevezal (1996,
156) warns of errors in carnivore bone ageing “due to the
high sensitivity and short period of persistence of the
record.” She also notes a number of the studies, mostly
on mustelids, showing better results with cementum. On
the other hand, the errors reported (except in some
mustelid studies) are more “margins of error” than totally
wrong (Klevezal 1996, Table 5). In addition, weasel
growth lines were not seen at all in one study (Klebanova
and Klevezal 1966) and reported in a study of British
weasels to have been egregiously affected by body weight
(King 1979). Stoats seem to stop at three growth lines
(Klebanova and Klevezal 1966) or are perhaps no good
for ageing at all (King 1991, 31). Suggested reasons for
the difficulties are remodelling and winter feeding habits
(Klevezal 1996, 96), the latter being supported by the
fact that hibernating animals produced clear winter lines
of arrested growth. Larger carnivores, like large mammals
in general, have had few reported studies, annual rings
supposedly not formed or observable in many of them.
While Klevezal does not mention studies done on
primates, she does in passing state that “apposition of
bone tissue in a human rib normally takes place at [i.e.
through?] the age of 60–69” (1996, 18).
Conclusion
Numerous hard tissues exhibit growth rings on various

21Using Osteohistology for Ageing and Sexing
scales, the most common being annual and the next,
daily. At least for most vertebrates, the best of these rings
seem to be on various dental tissues. Bones (and other
materials not covered in this paper) are also fairly effective
recorders of age. Because of factors involving specific
species behaviour, environmental variables, and differing
bone composition, growth and resorption; a simple
equation of major lines with years of age or over-
winterings and/or minor lines with months or days is not
possible. Instead, individual species and macro-
environments must be considered in order to accurately
read the lines. Fortunately, a lot of work has been done
so that, with adequate training, the archaeozoologist can
age remains of scores of species using growth rings. On
the other hand, even in well-researched taxa, there are a
number of problems that might be encountered. In
addition to the limitations listed at the end of the section
on dental growth rings, there are the following similar
restrictions on bone: the growth rings are often absent
for whatever reason; climatic conditions can mask lines
or conceivably induce extra ones. Resorption is a problem
on most long bones of most large and fast-growing
animals; missing, multiple, and unclear lines can lead to
minor or even major errors in counts. Finally, of course,
the bones or parts of bones that are needed can also be
missing or in a bad state. Where these problems are
present, other histological approaches might be effective.
Still, it has been noted that there are a number of
species where growth rings have already been shown to
be effective ageing devices. Some of the rings occur in
bones and teeth that are sometimes found in great
abundance at archaeological or paleontological sites.
From Peabody (1961) through Klevezal (1996), research-
ers have noted the impact of the environment on the
width of growth bands. If a large enough sample is
present, it stands to reason that it could be used to set up
a local or even regional floating dating sequence,
analogous to early dendrochronologies. Not only could
such a sequence be used for dating, but specimens that
have broken records could be matched to the master curve
and thus be aged. This idea might seem far-fetched, but
it seems, at least theoretically, possible, and striving for
it might refine our tools.
Osteons
As already mentioned, growth rings in bone and teeth
can be problematic for ageing. Therefore it makes sense
to look at other histological indicators. So far, this means
osteons and their environments. Over sixty years ago,
Amprino and Bairati (1936) noted specific changes in
bone histology of animals as they grow older, and in
their encyclopedic study of vertebrate osteohistology,
Amprino and Godina (1947) made cogent generalized
observations about the trends in bone fine structure as it
grows older as did Filogamo (1946) and Enlow (Enlow
and Brown 1956–58; Enlow 1963). While the specific
changes vary with bone and taxon, there is a general
pattern of three stages: 1) growth, 2) growth and change
(remodelling), and 3) regressive change (remodelling).
Schmidt et al. (1981) applied these stages to human bone,
but they have been shown to be applicable to many or
most species, especially longer-lived ones. More
specifically, bone modeling begins in infancy with no
secondary osteons, followed by secondary osteons that
are usually large and irregular as remodelling begins.
Small canals form in “youth,” followed by smaller osteons
and increased fragments in adulthood. Finally larger,
often unclosed osteons form with increased
numbers of smaller interstitial (i.e. fragmentary) lamellae,
and large resorption lacunae in individuals that reach an
old age (“senility”). While Amprino’s observations form
the basis of the qualitative method for ageing Homo
sapiens, no concerted effort has been made to produce
equivalent refinements in more than a few other species
even though informal use is sometimes made of this
knowledge (e.g. Lind 1994). Likewise, the quantitative
methods that have been developed for humans have had
almost no testing in other species.
Humans. Efforts at using osteon characteristics to deter-
mine age-at-death have concentrated on humans. The
approaches can be divided into two classes: quantitative
and qualitative. The former approaches, developed in the
English-speaking world and also used in many other
countries, attempt to count and measure any number of
variables. These include size, frequency, and relative area
of secondary and primary osteons, interstitial lamellae,
and canal diameters in pre-specified locations on most
long bones, ribs, and, less commonly, the mandible and
skull. These measurements are then matched with the
values for known ages-at-death to come up with precise
correlations and a statistical range. For a survey of most
of the numerous quantitative methods as well as a general
discussion of this class of ageing, see Robling and Stout
2000. The variables usually used are non-linear in their
distribution over age. Thus, simple single-variable cor-
relations or linear regressions, though giving values that
for some samples are within three or so years, are
inherently limited. On the bones, samples from midshaft
are generally advised. Because osteon distribution is not
symmetrical around the bone and counting is time-
consuming, and since collection administrators do not
always allow removal of a complete chunk of bone,
sampling procedures have been developed or, perhaps,
“evolved.” Thus, it is generally recommended that four
or more samples be taken, located equidistant around the
shaft, extending from periosteal to endosteal bone. The
most effective (i.e. accurate and precise) factor combin-
ations, as well as the best bones and loci cannot really be
specified, since the techniques have generally only been
“tested” on the same material – even the same data –
from which they were generated. In addition, different

22 K. Dammers
ages appear to vary in factor effectiveness. There are a
number of technical difficulties with these quantitative
approaches. Some of the problems are ameliorated by the
qualitative method explained below. In counting or
measuring osteons, what constitutes an osteon can be a
problem. While this seems like a resolvable problem when
one reads the procedures, a look at actual slides reveals
that identifying osteons can be quite difficult. Even with
image-processing software, there are difficulties and
ambiguities. A few of the studies have examined intra-
and inter-observer reliability, but a recent article
(Lynnerup et al. 1998) has reported discouraging statistics
in this area. Even though semi-automatic software has
sped up the counting and measuring, it has not resolved
the accuracy issues. Another significant problem deals
with loci. In trying to be objective and consistent in using
a limited sample, researchers can be confronted with
unacceptable material (i.e. bone that has taphonomical,
physiological, or pathological anomalies), and the suit-
ability of the bone may not be known until after the
sampling. Bone is often not expendable, and a further
sample might not be available. A final difficulty is
familiar to zooarchaeologists: the bone one is presented
with is not necessarily what one would consider ideal.
For the histologist, the problem is not the fragmentary
nature, but bones or bone regions for which no good
correlation between histology and age exists. Fortunately,
however, central parts of long-bone shafts are the ele-
ments most likely to survive archaeologically and resist
fungal and other modifications; these are the preferred
objects for histological ageing. A last and final impedi-
ment is that the condition of the macro-states can belie a
totally different degree of preservation on the cellular
level. On the other hand, the moderate independence of
the state of the two levels means that we can switch from
one to the other on occasion to get more information.
The qualitative method for ageing, developed in
Göttingen and practiced in Germany and neighboring
countries, takes a holistic approach: A number of factors
are weighed to get an age range. While this might at first
seem subjective and requiring extensive training, ageing
this way can in fact be learned quite rapidly. While I
know of no formal studies testing for consistency, in
practice people with just a few hours training generally
produce consistent ageings, a condition maintained over
years. While a non-lab description of the method is not
enough to make one a practitioner, it can give a good
idea as long as one follows up with hands-on work.
Perhaps the key to the qualitative method is its flexibility
in finding an appropriate area of the bone to use. The
general location, i.e. midshaft on long bones, is the same
as for the quantitative method, but the worker is not
forced to use exact loci that might be damaged or hard to
read. Furthermore, by using an impressionistic approach,
this method is quick and maximizes the human ability to
recognize patterns and tendencies while taking into
account anomalies. Fig. 8 shows the six major age
categories in their idealized forms. These stages can, in
turn, be subdivided to reflect transitional states, in effect
give ageings often both closer and surer than those
produced by the quantitative methods. A recent discussion
of the latter notes that they “all use linear regression
equations, which are notoriously limited…. Confidence
intervals are neither small nor equivalent but expand as
the regression line moves away from the mean” (Meindl
and Russell 1998, 387; no mention is made of qualitative
studies). The qualitative method has been found effective
with small, burned bone fragments as well (Hummel and
Schutkowski 1993). Recently, “super osteons,” giant
canal systems that occur away from midshaft, have been
examined and reported to increase fivefold from 20 years
of age to 80 (Bell et al. 2001a). Using super osteons thus
appears to be a potential additional ageing index. A
modest number of other animals have been examined for
osteon or general histological change over age. The
following is a sample of some of the more interesting and
relevant cases.
Primates. Osteohistological studies on primates of known
age have produced modest to promising results for
predicting age using factors involving osteons. The bone
turnover in female rhesus macaques (Macaca mulatta) is
clearly different for growing, adult, and menopausal
individuals, according to Colman et al. (1999), though
no specific mention of osteons is made in their paper.
Przybeck (1985) studied the osteohistology of the ribs of
15 macaques (sex not indicated), ranging from 4 to 31
years of age. Results of his limited sample suggest that
intact osteon density and osteon fragment density can
produce a good qualitative scale. For the youngest
animals, intact osteons are more diagnostic, while frag-
ments appear to be more useful in mature individuals.
Havill (2004) recently reported an increase in femoral
osteon density with age in her sample of 75 free-roaming
rhesus monkeys. A statistically significant, relatively
linear correlation showed age accounting for 23% of the
variation in density. Bowden et al. (1979) found only
extremely weak correlations between age and histological
factors for pigtail macaques (Macaca fascicularis), with
femoral osteon density showing a tendency to decrease
with age, though anomalous individuals clouded the
statistics (Table 27–4 on p. 341). What does come across
in these results is that “variation in degree of remodelling
[…] clearly increased in the older age groups […]” (342–
343). The authors also pointed out an easily noted increase
in the number of partially mineralized osteons and
irregular borders of osteons (p. 343), indicating that a
qualitative scale might work. More recently, Burr (1992,
187; see also Burr et al. 1989) found in a study of femurs
of 54 individuals that “[v]ariation in bone microstructure
occurs independent of gender, age, and body weight in
growing macaques.” On the other hand, he does note
that the rate of osteon accumulation in growing macaques
(but not in most other primates) seems to be stable and

23Using Osteohistology for Ageing and Sexing
that osteon population density is greater in adults (Burr
1992, 185). Lees and Ramsey (1999) noted differences in
the histology of the iliac bone of young (3–4 yrs.), mature
(10–15 yrs.), and old (22–24 yrs.?) cynomolgus monkeys,
saying that the results were similar to those for rhesus
monkeys. Thus, at least a distinction between growing
and adult macaques seems to be possible. Schaffler and
Burr (1984, 192–193) reported fifty percent more (14.6
vs. 9.7) osteons per millimeter in the femur of an adult
spider monkey (Ateles fusciceps) than in that of a sub-
adult (A. paniscus). On the other hand, there was no real
difference (7.7 vs. 7.4) in the values for an adult and a
subadult Pan troglodytes (Schaffler and Burr 1984, Table
1 on p.192). Mulhern and Ubelaker (2003), on the other
hand, found moderate age-related changes in the micro-
structure of juvenile chimpanzees (N=12) and significant
Fig. 8. Qualitative osteology for ageing human bone: generalized drawings of six primary stages of long bones in cross
section (modified from Hummel 2001, 32).

24 K. Dammers
changes for the one 35-year-old. For juveniles, age
“predicts 9–44% of the variation in chimp femoral
microstructural variables” (2003, 130). The clearest
indicator was the number of non-Haversian canals, which
decreased with age. Unfortunately, no indication of sex
is given in this study. The differences between mature
and old were dramatic.
Since primate osteohistological studies tend to focus
on locomotion and on contrasts with humans for use in
medical research, some bones that might be useful for
ageing have not gotten adequate attention to determine
their feasibility.
Carnivores. Examining images of mandibles from Arctic
fox, sable and mink in Klevezal and Kleinenberg (1969),
one can see that an increase in the density of osteons
takes place as these animals age. Stanek (1970)
specifically observed the rate of osteon growth in young
dogs.
Ungulates. In hog, peccary, cow, ox, and goat, there are
well-organized plexiform structures in the periosteal
bone. “In the formation of this bone, lamination of
vascular canals occurs […] connected with short, radial
canals” (Enlow and Brown 1958, 204). Metatarsals of a
young and an old rhinoceros (Amprino and Godina 1947,
Figs. 90 and 91) had the following suggestive differences:
secondary osteons of various sizes as well as fragments
are already present in the young animal but become the
only kind in the adult, where they are even more dis-
organized in the cortical core. By comparing the osteons
in the mandibular diastema of young and old wild Asiatic
asses, we can see that they become more compact and
appear to have cement lines and to become less layered
(Klevezal and Kleinenberg 1969, 101 [Fig. 61]). The
investigators note specifically: “in the young animal, the
main bulk of the buccal [cheek] wall consists of reticular
bone tissue and bone plates divided by many canals giving
passage to blood vessels; on the exterior we observed a
very narrow periosteal band without adhesion lines [...].
The lingual wall was completely osteonized. In the old
specimen, both the buccal and lingual walls were fully
osteonized” (Klevezal and Kleinenberg 1969, 102). In
the axis deer, there is rapid resorption. In the illustrations
in Klevezal and Kleinenberg (1969, 107, Fig. 66), the
difference between the two mandibles (six years and eight
years) is so great that I am inclined to think that it reflects
a special case. However, if it is not idiosyncratic, this
rapidity of change would make for easy ageing. Jane
Ruddle (1997) explicitly tested the applicability of the
quantitative approach as performed on humans, to roe
deer. Using a sample of mandible rami from 30 bucks
and 42 does, up to 14 years old, she found that the total
and average non-Haversian canal counts correlated highly
with age, the latter having an average inaccuracy of 2.5
years and an average bias of -0.04 years. This is not as
good as tooth-wear ageing, but of value where dentition
is absent, she concluded. This success with a moderately
long-lived and archaeologically significant animal is very
encouraging.
Rodents. An early quantitative examination of rat histo-
logical change showed that the pattern in these short-
lived mammals is close to that of humans. In fact, these
are the most dramatic results of osteon ageing. Using a
control laboratory sample of 35 rats from 2 to 120 days of
age, Singh and Gunberg (1971) found very good agree-
ment for femur, tibia, and mandible. Readings on the last
of the bones provided an accuracy of ±1.5 days for 67%
and ±3 days for 95% of the population’s true ages using
a simple regression based on increasing number of
secondary osteons, lamellae and “Haversian canal dia-
meter and non-Haversian canals” (250). Thus, specific
histological changes in rats are in agreement with the
known general osteoid maturing pattern of more osteons
forming with ever-more constricting rings – but for this
species, to an extent, that is outstanding. A cautionary
note: since the rats were from a controlled, single-sex
sample, variation is almost certainly less than what occurs
in nature.
Lagomorphs. Rabbits have a histological make-up dif-
ferent from that of rodents. “The vascular pattern […] is
longitudinal, and the structural arrangement and form-
ation of the primary osteone is not comparable [to that of
rodents]. In the development of new primary bone, the
deposition of periosteal lamellar layers alternates with
vascular strata. Each primary canal is surrounded by one,
two, or three rings of osteocytes forming a Haversian-
like structure. These primary vessels may extend across
the greater portion of the compacta without secondary
intervention, or areas of dense Haversian reconstruction
may occur. The ribs and skull bones are largely Haversian
structure, and well-developed secondary osteones appear
in several generations” (Enlow and Brown 1958, 211). A
quantitative study (Korkmaz et al. 1996) found a good
(r>0.9) correlation between increase in number of osteons
and age in the long bones (femur, tibia, humerus, ulna,
and even radius) of Lepus capensis. Also increasing with
age were what they called “interval lamellas” (presumably
interstitial lamellae), while non-Haversian canals de-
creased with age.
Sea mammals. Klevezal and Kleinenberg (1969) report
for the mid-mandible of the sei whale (Balaenoptera
borealis), that “macrocircular laminas [are] osteocized,
in small [i.e. young] specimens less so than in large
ones” (85).
Marsupials. Amprino and Godina (1947, Figs. 54 and
60) show that primary bone, with small vascules, is
replaced with the spread of secondary osteons, but not to
the periosteal surface. They also note that there is a
development of extreme disorder and that fragments help
separate islands of primary bone.

25Using Osteohistology for Ageing and Sexing
Avifauna. Except for aspects related to the need for light-
weight bones, the osteohistology of birds is similar to
that of mammals (Hodges 1974). Unfortunately, wild
birds seem not to have been studied for correlations
between age and changes in general bone histology.
Domesticated chickens have been fairly well studied. In
their early pre- and post-hatching development, their
Haversian canals, overall histological structure mineral-
ization, and cortical width are outstanding age markers,
differing from day to day, but only up to two to three
weeks post-hatching (Caplan 1988; Williams 2000). In a
recent survey of the literature on the histology of birds,
Rensberger and Watabe (2000) stated that “[l]amellar
bone is scarce in birds and coelurosaurs, and where it
does occur, it is poorly organized” (621).
Reptiles. Enlow (1969) describes a variety of general
developmental changes in reptile histology (59–66)
roughly like that of mammals, including one peculiar to
crocodilians and turtles that further studies could likely
stratify into age groups. Certainly, there are clear dif-
ferences between neonatal, young, and mature that can
already be designated. In fact, for some dinosaurs and
other early reptiles this has already been studied
(Chinsamy 1995; Horner et al. 2000; Ray and Chinsamy
2004). Here are some specific modern reptiles: Slider
turtles have a clear pattern of cellular development, with
males and females fairly similar but with a lag in female
bone and some variations depending on location on the
bones. Specifically, in males of the length of 65–100
mm, by the time of attaining the maximum length the
“vessels within the cortices were primarily longitudinally
arranged with some radiating communicating canals of
Volkmann” (Suzuki 1963, 357). This is followed by a
transition “to an outer primary longitudinal vascular bone
with an inner endosteal Haversian bone” (357). The Nile
monitor lizard’s tibia goes through a decrease in the
number of vascules, which also become radial in arrange-
ment and larger (Amprino and Godina 1947, Figs. 5–7).
It is possible that there is an increase in order, contrary
to what happens in vessels (osteons) in many other
animals. While most of the emphasis on dinosaur hist-
ology has been on lines of arrested growth, a number the
researchers have also noted and discussed differences in
osteons and other microscopic bone structures when
comparing younger and older individuals, e.g. Erickson
and Tumanova (1995, 2000), who in one species found
numerous stages of vascular patterns, from longitudinal
through reticular to radial, rather than the usual
Haversian systems of other dinosaurs (Reid 1997b).
Amphibians. The bone microstructures of very young and
adult frogs differ. For example, in the young bullfrog the
bone “is nonvascular, but subsequent peripheral de-
position results in simple vascular bone tissue” (Enlow
and Brown 1958, 214). The potential here for using
general histological structure to at least develop age
groups seems to be quite good.
Conclusion
There are certain general patterns that pertain to the
changes in osteonal bone with few exceptions regardless
of the bone and species, as long as that bone type is
present in a species. The extent and rate of these changes
vary markedly. What is needed is a series of studies of
local samples of various species with known ages and
sexes to develop fine-scaled charts of the specific nature
and rate of these changes.
Conclusion
Up until now, histological research on ageing (especially
for bones) has concentrated on the so-called second order,
the scale of the growth lines, osteons and lamellae
themselves. The tools have been simple microscopes to
examine two-dimensional images of cross-sections. Hist-
ology of non-lamellar bone has been all but ignored when
it comes to attempting age determination. This is now
changing, or at least being augmented by new tools that
can image non-invasively or on finer scales, with auto-
mated programs and ones that produce three-dimensional
images.
Sexing with Osteons
There has been little direct work on sex determination
using osteons or other histological landmarks. This
paucity is likely due to the fact that there are easier and
presumably more effective methods in osteology. What
work has been done is characteristically indirect, with
the researchers usually looking at sex-related factors or
simply controlling for sex. Indirect studies can be put
into five groups: empirical studies, mechanical stress,
disturbances, female functions, and the third sex. Some
early osteonic studies disregarded sex, and others claimed
that it made no difference. Later studies showed that sex
does statistically make a difference, even in Homo
sapiens, a species with only moderate sexual dimorphism.
Thus, the more careful quantitative studies now do
separate by sex, so as to get cleaner results.
Empirical Studies
An innovative study (Smitmans et al. 2000) using SAM
to examine the acoustic impedance of lamellar osteon
ensembles of a sample of human material covering a very
wide age range also compared differences in sex. While
impedance generally increased over age with no particular
difference between the sexes, there was reversal of this
trend in the oldest males. The decline is small and
gradual, but for either a large collection or an older range
group, SAM offers some promise for sexing well-aged
animals that live to the advanced age category. However,
work has only been reported on one species so far.
Another, clearer difference was the one-third higher rate

26 K. Dammers
of young osteons in women than in men (Bell et al.
2001a, b). In a work that is difficult to obtain, Egeli
(1976) reported on the differences in osteon sizes in men
and women. While both boys and girls exhibit a con-
tinuous decrease in osteoid surface extent as they grow
older (from 34% before 6 yrs. to 17% by 23), the rates
vary by sex (Glorieux et al. 2000). Given the tight ageing,
this discrepancy might fairly be used to infer the probable
sex in young humans. The same study also found differ-
ences in osteoblast surface extent between the two sexes.
Burr (1992, 187) found bone microstructure of no
predictive value for determining the sex of growing
macaques. On the other hand, Havill (2004) reported that
osteons in mature males are larger than those in mature
females. While she points out that the size difference can
be explained by bone (or body) size, for archaeological
samples it can be used as an indicator of sex.
In both raccoon dogs and badgers, a number of differ-
ences have been recorded between the sexes, including
osteon short diameters and osteon density (Hidaka et al.
1998). Osteon diameters in humans have also been
reported as being greater in adult males than in women
(Brockstedt et al. 1993), suggesting that this factor could
discriminate in a number of species. Klevezal (2001)
reported an accuracy rate of sexing Apodemus mice
mandibles histologically at over 80%.
Suzuki (1963) noted that male slider turtles have a
thick cortical wall and massive Haversian canals in the
endosteal region, in contrast to females. This species
also offers a good example of a sexual difference in age
of microstructural change, as mentioned in the ageing
section above, that can provide information for sex
determination.
Mechanical Stress
Wolff’s Law states, somewhat hyperbolically, that the
bone responds to loci of stress. In both bone build-up
associated with muscle attachment and use, and in impact
stress, the changes are actually activated on the micro-
scopic, chemical level, with the living bone bringing up
reinforcements to provide improved infrastructure. It is
almost as if the body knows what it will need and plans
for it, whereas how it “knows” and exactly how the
mobilization takes place is not understood by scientists.
As we know, within a given population, human males
tend to develop larger muscles and more robust and denser
bones; some corresponding differences are seen in the
osteohistology. The same can be hypothesized for other
sexually dimorphic species (or reverse, for species with
more robust females). Histomorphologically, many of the
distinctions based on robustness in macromorphology are
observable less clearly, but with the same exceptions and
overlap.
Disturbances
The most common disturbances are injuries and diseases.
Histological analysis of disturbances associated with a
particular sex can, like macromorphological studies,
suggest the sex from bones of the individual from the
microscopic changes if they are present. Beyond the
influences of disease and injury are the long-term be-
havioural ones. In humankind, we have the tennis elbow
(which does not show the expected histological change)
or the well-known grain-grinding activity often associated
with women in Neolithic cultures (I do not know of any
studies on the histology here, as opposed to those on the
macromorphology). But there are also behavioural differ-
ences in other species as well: consider the differences in
hunting styles of the lioness and her male counterpart or
the differential use in many cultures of cows vs. bulls and
stallions vs. mares, or primate and marsupial carrying of
young. This issue, unfortunately, has hardly been ex-
amined.
Female Functions
It is not surprising that osteohistologists, in general, focus
on females. The physiological effect of hormonal changes
and rhythms are more obvious than in men, though the
extent to which the influences are direct is still far from
clear. Thus, menarche, menstruation, pregnancy,
parturition, lactation/nursing, weaning, and menopause,
each have been associated with changes in resorption as
seen in histological studies in various species. Physio-
logically, these associations are not surprising: each of
these phenomena makes a significant and different
demand on the body’s resources, especially the calcium
and iron. Unfortunately for sexing purposes, many of
these changes are transitory. For other changes, some
permanent, see the study of parturition-lactation zone in
the cementum of souslik teeth in Trunova et al. (1999),
as well as Klevezal and Sukhovskaya (1995) on mice
teeth, Rosenberg and Simmons (1980) on rabbits and
pregnancy, and Kagerer and Grupe (2001) on a cor-
relation between bright lines and pregnancy/parturition
in women’s cementum.
Menarche/Sexual Maturity
In slider turtles, the females’ osseous changes take place
at older ages than do those of the males. Thus, if the age
can be determined – e.g. by scute annuli – the sex should
be histologically determinable in individuals whose
development of ≥65 mm carpal length has not yet
plateaued. For example, male turtles with carpal lengths
of 65–100 mm have a much heavier layer of endosteal
Haversian canals than do females of the same age (Suzuki
1963).
In humans, sex differences for changes in bone micro-
structure have been reported on two occasions. The first

27Using Osteohistology for Ageing and Sexing
was the age of onset of resorption patterns in the cortex
of bones of teens, with peripheral resorption coming
earlier in females. This earlier resorption occurs around
menarche, either due to the accompanying change in
physiological stress or associated with the general
pattern of earlier maturation we know in girls. Glorieux
et al. (2000, 108) suggest that histological sex
differences in girls “might be the result of differences
in the timing of puberty.” If a subject can be age-dated
to within a year or so and the age falls within the early-
to-mid-teens, this fact can be used to sex the individual.
Usually if we have enough data to age an individual
that accurately, we have enough gross morphology or
DNA to sex with greater probability of accuracy than
histomorphologically, so the point might seem moot.
However, this procedure might find better application
in other species, ones that exhibit or suggest appreciably
different onsets of sexual maturity, or something of the
kind, between the sexes.
Reproduction (pregnancy, parturition, lactation)
Chamberlain and Forbes (2001, in press; see also Forbes
1994) found a clear difference in distribution of osteon
size in femurs of beef cattle (male and female) and of
dairy cattle (obviously female). The dairy cattle osteon
size distribution was statistically identical to that of cattle
from a Roman site in England from around AD 100. A
diagnostic pattern seems to exist, though it does not have
to be sex-related per se. Unfortunately, whether it is due
to behaviour (extensive/intensive care), diet, breed,
dairying/lactation/milking, or whether it is a reflection
of age, is not clear since the dairy cattle were all much
older than the beef cattle (2 yrs.). Since remodelling and
differences in osteon size and density are related to at
least both lactation and age, this study has opened a door
to sexing techniques (Saddha Kuippers, pers. comm.). In
any case, there is an osteonal difference between young
beef cattle of either sex and older milch cows. Whether it
is due to the hormones or the treatment (or extraneous
factors), it is possible that this study will lead to
identifying sex if we know that in a given culture milked
cows were the only cattle kept to an older age.
Since lactation puts a strain on the calcium supply, it
is to be expected that in most species (unlike rats, which
are efficient enough calcium producers to handle this
strain with ease), there will be a histological effect from
nursing. Transient increases in intracortical bone re-
modelling reflects the drain on the mother’s calcium
resources. Lactation in beagles, for example, strongly
remodelled the bone, with a significantly different make-
up in osteon composition and resorption spaces (Vajda et
al. 1999, 1441–1443). These dogs, however, were from a
highly controlled lab population, so the results are probably
clearer than would occur in nature. It should also be
pointed out that this and almost all female function studies
use other females and not males as controls. The fact that
the change is transitory also reduces the diagnostic value
of lactation changes. Vajda also notes studies that have
shown a correlation between lactation and changes in
cancellous bone in sheep, pigs, monkeys, and other
animals, “[h]istomorphometry revealing that the lactation-
induced decrease in bone mass is associated with a marked
increase in cancellous bone remodelling as reported in
dogs… and rats…” (Vadja et al. 1999, 1439).
Another study (Ott et al. 1999) looked at macaques.
Osteoclast density increased during pregnancy and was
still present three months after weaning. These results
are quite promising, though the variability was very large.
An archaeological report by Trivers and Armelagos
(1977) suggested that a hypermineralized ring directly
around the Haversian canal or slightly farther out de-
creases with lactation in humans. Thus, a small variety
of effects on osteons has been reported associated with
reproduction and lactation. To what extent the effects of
demands on the mother’s system, mostly for calcium,
repeat across species (or even samples) is not known,
since the variables measured differ from study to study.
Menopause
Resorption of medullary bone in middle age begins earlier
in women than in men. The onset in women might be
linked to menopause. One could hypothesize a similar
difference in age of changed resorption pattern for other
long-lived mammals, such as horses, deer, dogs, cats,
wolves, monkeys, apes, and elephants. In humans, ageing
on undocumented individuals is too inexact to make this
slight shift of use in sexing individuals. However,
statistically the difference is enough to calculate a popu-
lation sexual distribution given a large enough maturens
sample. In any case, bone mineral density and turn-over
are well documented in menopausal females, including
in monkeys (e.g. Champ et al. 1996; Colman et al. 1999;
Cerroni et al. 2000) as well as humans, even though the
work is generally not histological. On the other hand,
dental growth rings seem to be accurate enough to enable
the use of this shift for sexing (my speculation); likewise
for species where bone growth rings are present and
complete enough to allow for close ageing. Late mature
individuals and ones with greater resorption than
“expected” for their closely determined age could thus be
sexed for any taxa also exhibiting the sex difference in
age of middle-age resorption onset.
The Third Sex
Certain domesticated animals are castrated in a number
of cultures. The surgery can be undertaken for a number
of purposes: to improve docility, to control populations, to
fatten animals for meat, or to control singing voice (in
humans). It stands to reason that this massive modification
of the hormonal system would leave its mark on the
microscopic structure of hard tissue.
Castration has been shown to affect the development
of the histology of antlers, generally producing a juvenile

28 K. Dammers
appearance, with a lack of secondary osteons (Kierdorf
et al. 1995, 38–39, 41; Bubenik 1990, 281–283).
Castration also affects the structure of bone, generally
reducing the bone density (Winks and Felts 1980). In
rats, the accompanying structural changes include a
sparcity of trabeculae and an obvious increase in re-
sorption cavities in femurs. However, the mandibles were
unaffected in all aspects studied. The difference seems to
appear more in load-bearing bones. Another contributing
factor affecting histological changes is the age at which
castration is performed. Strangely, animals castrated at
one year but not those operated on at twenty-three days
exhibited change in bone structure! Since (early)
castration in larger male animals can apparently alter
the morphology of some bones (e.g. bull metacarpals; the
pelvis) to the point of shifting them into the range for
females (Chaplin 1971, 103–104; Deborah Ruscillo, pers.
comm.), it is possible that histological studies could also
show an analogous shift for them.
Summary
Thus, a number of differences occur either incidentally
or inherently in the microscopic bone structures of the
sexes. However, no special effort has been made to
compare them across the taxonomic landscape. While
space restrictions have essentially limited my discussion
on sex to osteons, in fact, bone and tooth annuli also
reflect many of the same forces. The interested reader is
directed to Laws (1953), Spencer and Uhler (1955, 75–
82), Klevezal and Kleinenberg (1966, 116), Carlson
(1990, 543), Klevezal (1996, passim), and Klevezal
(2001). When only small fragments are present, macro-
morphology is often stymied. Furthermore, the best bones
for morphological and histological sexing are not the
same. DNA sexing is still expensive and not always
effective, e.g. in some cases of burning, where osteons
can be quite robust. In truth, though, it seems there will
be only some occasional cases, such as dog or human
cremations with teeth remains and bone splinters, where
histological sexing will prevail.
Conclusion
We can envisage a continuum from the macroscopic/
morphological over the histological to the chemical and
molecular. There is a rough corollary with this ever finer,
ever smaller focus: the less visible the objects of study,
the longer it takes and the more complicated and ex-
pensive it is to get results. Osteonal and other histological
studies have several advantages, perhaps foremost of
which is the generally robust character of the osteons:
small, broken pieces of burned bone are adequate material
(Grupe and Herrmann 1983/84; Hummel and
Schutkowski 1986; Cattaneo et al. 1999); and it is the
strong shafts of long bones, along with some cranial
material, the most persistent of bones, that are best suited
to osteon analysis. An additional useful feature is that
once the thin-sections have been prepared for ageing,
say, no further preparation is necessary for tests of stress,
disease, and sex. In fact, these tests often should be
undertaken together to get the most accurate results.
Additionally, the thin-sections are small and generally
long-lived. To a certain extent, the osteohistological
methods developed to date are complementary to each
other. This is often true in terms of species and bones as
well as the obvious (though small) difference in location
on the bones. On the other hand, some of the taphonomic
depredations affecting one affect the other.
Obviously, both grosser and finer methods also have
certain advantages as well. The decision as to what
particular tool or tools to use depends on the nature of
the remains and the questions to be asked (as well as
funds and time), and should not be determined just by
the availability of an expert or even the lack of knowledge
of the existence of alternative methods and techniques.
In some cases, a combination of methods can bring better
results than individual techniques used separately.
Furthermore, there is still enough mystery in animal
histology that such studies can profit from such co-
operation. As David Browman put it in another context
in Early Native Americans (1980, preface), we need to
“attempt to breed hybrid vigor by mixing a variety of
approaches and directions together.” While a fair amount
is known about osteons, especially in taxa determination
(e.g. Foote 1913; Mátyás 1927; Amprino and Godina
1947; Enlow and Brown 1956–58), ageing of Homo
sapiens (Pfeiffer 1992; Robling and Stout 2000), a bit
about aspects of domestication (Richardson et al. 1961;
Lasota-Moskalewska and Moskalewski 1982; Gilbert
1989; Forbes 1994; Chamberlain and Forbes 2001, in
press), and certain aspects of stress (Burger et al. 2001;
Skedros 2001) and disease (Pfeiffer 2000); more focus
needs to be put on the usual suspects – cows, pigs, goats,
horses, dogs – as well as deer, birds, and other common
game and commensal animals, especially in terms of
ageing.
Acknowledgments
I would like to thank Deborah Ruscillo for care in
organizing the ICAZ ageing and sexing session and her
support and help throughout the session and editing this
volume. I also thank Jaco Weinstock and Elaine Turner
for their critical reading of an earlier version of my paper.
Bryan Gordon, Wim Van Neer, and Darius Batulevicius
provided helpful advice. Andrew Chamberlain graciously
allowed me to use a number of definitions from the web
site of University of Sheffield’s Department of Anthrop-
ology. For generously granting me the free use of
illustrations, I gladly thank the editors of the Turkish
Journal of Zoology as well as Gary Rosenberg and the
Springer-Verlag. I acknowledge the generous travel funds
to attend ICAZ 2002 granted by the Institute for Aegean
Prehistory.

29Using Osteohistology for Ageing and Sexing
Glossary
N.B.: The definitions given here are aimed at providing simple
and understandable descriptions to help in following the
literature rather than authoritative definitions (see Gordon
1993). For the latter, see, e.g. Carter 1990; Francillon-Viellot
et al. 1990; Parfitt et al. 1987; and Report of the Workshop
1980, as well as standard science and medical dictionaries.
Italicized terms are defined elsewhere in the glossary.
Definitions marked with an asterisk (*) are from Sheffield, no
date:
http://www.shef.ac.uk/uni/academic/A-C/ap/thin_sections/
glossary.html (with possible orthographic changes), used with
kind permission.
adhesion line older term for recording structure or cement
line.
ageing determining of the age-at-death or, for living
individuals, the amount of time since (or before) birth. Bones,
teeth, etc. lost during life are aged to age-at-time-of-loss, i.e.
to the age at which the item rather than the individual died.
Ageing is to be distinguished from “dating,” which refers to
length of time before the present or before or after some external
date.
alkaline phosphatase a phosphate-releasing enzyme secreted
by osteoblasts that calcifies the matrix.
anastomosed (integrally) joined.
annulus (pl. annuli) a ring-like structure or part, e.g. on trees,
shells, teeth, and in bones (cf. growth rings). It is sometimes
also understood as a year’s ring. This can be confusing, since
there is nothing innate in their being the same. Some authors
(e.g. McKinley and Burke 2001) use the term to refer only to
regions of reduced centrifugal growth with few cells, and
contrast annuli with LAGs (no cell, opaquer in polarized light),
and both with growth zone. Erickson (1997) simply describes
annuli as thin layers of avascular bone fibres.
apatite calcium phosphate, the main mineral of bone.
BMD bone mineral density.
BMU (bone maintenance unit / basic multicelluar unit) the
package of various bone cells that carries out remodelling.
bone modelling “the acquisition and transformation of overall
bone form and shapes by differential variation of local growth
rates” (Francillon-Viellot et al. 1990).
bone remodelling see remodelling.
bone structural unit the tiny packet of new bone formed in
resorption and found in Howship’s lacuna.
bone turnover change in chemical and/or structural com-
position of bone.
BSU bone structural unit.
canaliculi (sg. canaliculus) fine channels through bone that
form an anastomosing network between the osteocyte lacunae.
In living bone the canaliculi are occupied by the cytoplasmic
processes of the osteocytes.* They run perpendicular to the
Haversian canals.
cancellous bone spongy bone containing interconnecting
cavities.
cement(um) a cellular mineralized fibrous organic material
that is laid down on the exterior of a tooth’s root, accreting in
bands, the most exterior of which are the most recent. These
bands are effective ageing indicators.
cement line reversal line. Cement line is usually reserved for
a reversal line occurring in an osteon. On the other hand,
Francillon-Viellot et al. (1990, 505) and Klevezal 1996, 18)
define cement lines as including both resorption lines and
resting lines.
circumferential lamellae (circumferential lamellar bone)
sheets of lamellar bone lying parallel to the periosteal and
endosteal bone surfaces.* Bone “composed of evenly spaced
bands, or lamellae, that run parallel to each other around the
outer part of the cortex. It appears as birefringent sheets in
polarized light and can be distinguished by its long, parallel
fibers. [It…] is a prominent feature of childhood” (Kerley 1965,
151).
collagen a [tough] fibrous protein that constitutes the principal
organic fraction of bone. * There are various types of collagen,
but all in the form of a triple helix. Collagen comprises over
85% of the mineral content of bone.
compact bone (compacta) bone which has few “open” spaces;
it includes most external surfaces. “The differentiated compacta
of the bones of mammals usually consists of a mesiosteal zone,
and also an outer and inner layer of generative plates” (Klevezal
and Kleinenberg 1969, 9). It makes up about 4/5 of bone mass
in humans. Cp. cancellous bone.
concentric lamellae lamellae in osteons: surrounding the
Haversian canal.
cortical bone dense bone, or all dense bone with a network of
parallel collagen; essentially, a synonym for compact bone.
demineralization laboratory removal of mineral (primarily
calcium) by treating with acid.
dentin(e) acellular (and generally mineralized) organic sub-
stance which accretes in the tooth’s pulp cavity, advancing
inward in layers. These resulting bands are effective indicators
of ageing.
diaphysis shaft of a long bone.
drifting osteon a type of secondary osteon found in some young
animals; the osteon moves sideways by osteoclasts eating away
the matrix on one side of a recently created osteon, while
osteoblasts fill in on the other side (Robling and Stout 1999).
endosteal referring to the outer surface of a bone; the opposite
of periosteal.
endosteal bone the internal surface of cortical bone, adjacent
to the medullary cavity.*
endosteum the one-celled layer which lines the medullary
(inner) cavity within the long bones. It is osteogenic.
epiphysis secondary ossification center found at the neck of
long bones.
fibro-lamellar compact bone “a complex of 1) woven or
fibrous, cancellous matrix of periosteal origin, and 2) a lamellar
matrix, centripetally deposited, forming the primary osteons
which ultimately fill the space originally empty in the woven
periosteal matrix. Vascularization is plentiful, although vari-
able, and evidence of growth cycles is generally rare” (Carter
1990).
fragmentary osteon(e)s (osteon(e) fragments) partial osteons,
the remains of osteons broken down by osteoclasts building
new (secondary) osteons.
growth layer a growth zone plus an associated annulus or
LAG (McKinley and Burke 2001).
growth layer group a repeated pattern of incremental growth
layers (Hillson 1986, 224), considered to represent one of a
number of temporal units, especially a year.
growth mark “any variation in growth recorded at the morpho-
logical or histological levels in any sclerified tissue…”
(Francillon-Viellot et al. 1990, 507), and occurring as a growth

30 K. Dammers
zone, annulus, or a rest line. Incremental growth layer.
growth ring any ring or line indicating a period of growth;
“refers to cases in which periosteal bone of Gross’s zonal type
(1934, 742, as zonare Periostknochen) is divided into tree-
ring like zones by the cyclical development of major resting
lines… or bands of dense bone termed annuli…” (Reid 1990,
21). Synonyms and near-synonyms include: incremental line,
winter incremental line, winter resting line, daily layer, zone
of hibernation, parturition-lactation zone, dental growth mark,
incremental growth line, circadian increment, polish line, bone
layers, line of arrested growth, annulus, adhesion line, and
recording structure. Use has not yet become standardized, so
that the same term can have different meanings from author to
author and a different term for the same concept will be found
from article to article. Where the author is dealing with the
physiology itself, s/he will usually explain her or his use
adequately for one to follow the article.
growth zone a layer of bone not characterized by slow or
stopped growth. It is translucent in polarized light (McKinley
and Burke 2001).
Haversian bone “Bone which has been largely remodelled
into secondary osteons” (O’Connor 2000, 6).
Haversian canal one of the minute canals which traverse many
dense bones in a wandering manner, generally following the
major axis of the bone. They convey nutrients (via blood vessels,
nerves, and lymph vessels) to bone through their subordinate
adjuncts including osteocytes. A Haversian canal is the center
of a given osteon (Haversian system).
Haversian lamellae lamellae (sheets of dense bone) around
Haversian canals which were formed during osteon formation
and accrete concentrically inward.
Haversian system osteon.
histology the study of the microscopic structure of biological
tissue and function; loosely, such structures themselves.
Howship’s lacunae the surface depressions on bone surface
left by osteoclasts where bone resorption occurs, located
between the osteoclast and the bone resorption surface.
hydroxyapatite a complex crystal of calcium phosphate crystal,
Ca10 (P04)6(OH)2, that makes up practically all of bone
mineral and makes it hard and stiff.
incremental growth layers “distinct layers parallel with the
formative surface of a hard tissue (dentine, bone, cement and
their subtypes) which contrast with adjacent layers” (Hillson
1986, 224). According to Hillson, this terminology is
standardized for marine mammals.
incremental line (IL) a general term for incremental growth
layer or recording structure, and often growth layers or and
annuli. However, its use is sometimes restricted to just lines
reflecting stoppage or slowing of growth.
inner circumferential lamellae lamellae found adjacent to
the endosteum, i.e. near the medullary cavity.
interstitial lamellae (interstitial lamellar bone) remnants of
older Haversian lamellae and possibly of inner and/or outer
circumferential lamellae.
lacuna See osteocyte lacuna.
LAG line of arrested growth.
lamella (pl. lamellae) any of the thin plates of fibre, found 1)
bundled centripetally around Haversian canals (Haversian
lamellae), 2) occurring as fragments of the previous type of
lamellae in compact bone (interstitial lamellae), 3) added
centrifugally (periosteal lamellar bone or outer circumferential
lamellae), or 4) added centripetally (inner lamellar bone or
inner circumferential lamellae) to dense bone.
lamellar bone dense bone with fine collagen fibres organized
into sheets of a few microns thickness (lamellae), within which
the fibres are parallel in orientation.*
laminae “bone with zonal ‘growth rings’[…;] bone between
successive vascular networks (sensu Foote 1916, 11)…[;] “bone
between successive ‘bright lines’ or successive layers of
periosteal woven bone if ‘bright lines’ are lacking [sensu Currey
1960…]” (Reid 1990, 23, 24).
laminar bone (laminar Haversian bone) bone in which the
osteons appear to be lying in layers. A classic case is found in
numerous dinosaur bones. According to Reid (1990, 23), Foote
included both zonal and fibro-lamellar tissues, causing con-
fusion in subsequent work.
laminar fibro-lamellar bone Reid (1990, 23) notes various
uses: laminar bone (Currey and Reid), laminar and plexiform
fibro-lamellar bone (de Ricqles), plexiform bone (Enlow),
laminare Periostknochen (Gross).
line of arrested growth (LAG) a line on a bone, tooth, etc.
representing a band of growth, in which the growth slowed,
characteristically causing denser deposits. The slowing can be
caused by stress or slowed metabolism. The centrifugal growth
(in periosteal LAGs) is said to stop. Thus, Erickson (1997, 4)
describes it as an avascular layer thinner than and having fewer
fibres than an annulus has. A LAG is a particular kind of
growth ring.
line of resorption a usually distinct, sometimes wavy, line
indicating the outermost margin of (extensive) bone
remodelling, i.e. the boundary between the mesiosteal and
periosteal zones. It is presumably always axial to any growth
rings that (still) exist.
marginal zone in an osteon, the outermost band, bounded by
the cement (reversal) line.
medullary cavity the central tube within the bone which houses
the marrow, responsible for generation of blood cells.
mesiosteal zone the intermediary part of cortical (compact)
bone. “The mesiosteal zone forms as a result of the recon-
struction of the primary bone tissue” (Klevezal and Kleinenberg
1969, 9). The osseous zone of cortical bone.
metaphysis the site of ossification, between the diaphysis and
epiphysis.
microtome an automated sawing machine that cuts off fine
slices. It is very easy to operate but quite expensive.
mineralized having had minerals deposited as a crystalline or
amorphous mass in a pre-existing matrix (bone or cartilage).
non-Haversian canal primary vascular channels “formed at
the time the surrounding lamellar bone was formed,
[…representing] areas of unremodelled bone” (Kerley 1965,
151–2). Essentially the same as primary osteon(e).
osteoblasts secretory cells responsible for building new bone
(lamellae).
osteoclasts large, multinucleated bone cells that tear down old
bone by secreting an acid.
osteocytes bone cells which have lost their ability to produce
bone material and are trapped within their matrix. These are
the remnants of the osteoblasts which form bone tissue as the
bone is developing, are located within/between the lamellae,
with canaliculi radiating toward the central Haversian canal
and other osteocytes in the osteon.
osteocyte lacuna [usually referred to as simply lacuna, pl.
lacunae] a round, ovoid or lenticular space within which an
osteocyte resides in living bone tissue.*

31Using Osteohistology for Ageing and Sexing
osteohistology the histology of bone; loosely, the histology of
animal hard tissue or at least of bone and teeth.
osteoid precalcified bone matrix composed mostly of collagen.
osteon(e) (Haversian system) a structural unit of bone
consisting of a central (Haversian) channel with blood and
lymph vessels, nerves, and connective tissue surrounded by
concentric cylindrical layers of lamellar bone of osteocytes in
an apatite matrix. Volkmann’s canals radiate laterally from
the Haversian canal, allowing distribution of the blood, etc.
on a finer scale. See secondary osteon and primary osteon.
Osteon(e) Population Density (OPD) “[(osteon number +
osteon fragment number)/mm2]: the number of secondary
osteons with intact Haversian canals, plus the number of osteon
fragments/square mm averaged for all observed cortical fields”
(Havill 2004).
osteon(e) fragments fragmentary osteon(e)s.
outer circumferential lamellae (outer lamellae) (sg. lamella)
the thin layers of bone laid down as the bone is formed. They
lie just below the periosteum and form the outer surface of the
compact bone tissue.
parturition-lactation zone an area of bone with a distinctive
pattern coinciding with parturition and lactation.
Percent Osteonal Bone “[(osteonal area/cortical area) × 100]:
proportion of the observed cortex occupied by secondary osteons
with intact Haversian canals” (Havill 2004).
permineralized fossils having their original pore space infilled
with minerals.
periosteal referring to the outer surface of a bone; the opposite
of endosteal.
periosteal bone the external surface of cortical bone: “the
outer generative plates [...] deposited as a result of apposition
activity” (Klevezal and Kleinenberg 1969,9).
periosteum vascular membrane covering the surface of a bone’s
shaft and having osteogenic potential. Osteologists sometimes
also use the term to refer to the surface of the bone itself.
petrification the process or condition of original material of
an organism being replaced with minerals after death.
plexiform bone bone in which the lamellae are grouped into
layers separated by well-developed vascular channels or by
bands of osteons.*
polarized light light waves whose plane of vibration is confined
to a particular orientation.* This is often an effective way of
viewing bone sections under a microscope.
primary osteon(e) usually a small osteon that is not bounded
by a reversal line but instead is surrounded by conformable
layers of interstitial or circumferential lamellar bone.*
Essentially the same as non-Haverian canal.
recording structure persistent layers of hard tissue that reflect
response to changes in the physiological parameters of an
organism as it grows or undergoes environmental impact.
(Klevezal 1996, 3). It includes both lines of arrested growth
and annuli. Replacing the designation adhesion line, this term
is a near synonym to growth ring or incremental growth layer.
remodelling the body’s resorption and reconstruction of bone
(or other dense tissue) through biochemical activity, especially
by osteoclasts and osteoblasts, resulting in a new or modified
shape.
remodelling line resorption line.
resorption the biochemical reworking of bone through
incorporating old structures into new ones. This is primarily
done by osteoclasts. See Parfitt 1993.
resorption lacuna (pl. resorption lacunae) volume in compact
bone which has been resorbed without subsequent replacement
by Haversian systems.
resorption line a wavy band about 1–2 µ across indicating the
edge of resorption either within an osteon or mediosteally,
where it marks the inner limit of still-existing centrifugally
growing growth rings. It is essentially synonymous with
reversal line and is sometimes also called a remodelling line
or cement line (q.v.).
resting line (rest line) a line, characterized by a smooth border,
reflecting a pause in centripetal or centrifugal growth, i.e. a
LAG (Francillon-Viellot et al. 1990, 505).
reticular bone a type of fibro-lamellar bone in which primary
osteons in lamellar bone are obliquely oriented and connected
to each other.
reversal line narrow zone of hypermineralisation that
demarcates the boundary between the termination point of bone
resorption and the initiation point of new bone formation.
Sometimes referred to as a cement line.*
ruffled border edge of osteoclasts facing the Howship’s
lacunae and wavy in shape.
sealing zone that area that to which an osteoclast attaches and
in which it creates a Howship’s lacuna.
SAM scanning auditory microscope.
SCN suprachiasmatic nucleus.
secondary osteon(e) an osteon that has developed in a re-
sorption space within pre-existing bone tissue; distinguished
by its interception of pre-existing lamellae and by its hyper-
mineralised border (reversal line).*
SEM scanning electron microscope.
sexing determining of the sex of an individual to whom the
tissue under consideration belonged.
skeletochronology (sclerochronology) “the study of [faunal]
mineralized tissue for the purpose of determining intervals of
time with reference to the organism being studied” (McKinley
and Burke 2001, 34). The “study of recording structures
apparent in mineralized vertebrate tissues” (Pike-Tay 1999,
297). (Note that these two definitions are not identical, the
former allowing for other histological ageing methods such as
those based on osteons and the latter not unless recording
structures be redefined. It should also be noted that
skeletochronology is usually restricted to vertebrates, and
sclerochronology includes – often exclusively – invertebrates).
super-osteon remodelling cluster with a central canal having
a diameter greater than 385 microns (Bell et al. 2001a, b).
suprachiasmatic nucleus (SCN) part of the brain that regulates
circadian rhythms.
trabecular bone cancellous bone containing interconnecting
cavities having “a precise three-dimensional spatial
arrangement which reflects mechanical forces acting on the
bone” (Francillon-Viellot et al. 1990, 491).
vascular canal small tube within bone that provides a pathway
for blood, nerve, and lymph lines. Varieties include the central
canals of primary and secondary osteons, as well as simple
primary vascular canals without surrounding lamellae.
Volkmann’s Canals vascular channels running perpendicular
to the periosteal and endosteal surfaces, connecting the vessels
in the Haversian canals and (in some cases) connecting with
vessels in the periosteum and the medullary cavity.*
woven bone highly vascularised bone tissue with coarse,
undulating, interwoven and randomly orientated collagen fibre
bundles and randomly distributed osteocyte lacunae; found in
embryonic and fetal bone, fracture callus, and in the medullary

32 K. Dammers
bone of egg-laying birds.*
References
Overviews
Histology
Bloom, W. and Fawcett, D. W. 1975. A Textbook of Histology.
Philadelphia, Pa.: Saunders. [Also available in 1994 edition]
Bourne, G. H. (ed.) 1972. The Biochemistry and Physiology of
Bone. Twelve volumes. London: Academic.
Currey, J. D. 2002. Bones: Structure and mechanics. Princeton:
Princeton University Press. [Comprehensive overview]
Francillon-Viellot, H., Buffrénil, V. de, Castanet, J., Géraudie, J.,
Meunier, F. J., Sire, J. Y., Zylberg, L. and Ricqles, A. de 1990.
Microstructure and mineralization of vertebrate skeletal tissues,
pp. 471–530 in Carter, J. G. (ed.), Skeletal Biomineralization of
Patterns, Processes and Evolutionary Trends. Two volumes.
New York: Van Nostrand Reinhold. [A very good and applicable
technical guide]
Pritchard, J. J. 1972. General histology of bone, pp. 1–20 in Bourne,
G. H. (ed.) The Biochemistry and Physiology of Bone, vol. 1.
Twelve volumes. London: Academic.
Morphometry
Parfitt, A. M. 1993. Morphometry of bone resorption: Introduction
and overview. Bone 14, 435–441.
Bone Growth
Spencer, M. and Uhler, K. (comps.) 1955. The Structure, Com-
position and Growth of Bone 1930–1953. Washington D. C.:
Armed Forces Medical Library, Reference Division. (pp. 131–
132: Structure – variations with age) [lightly annotated biblio-
graphy]
Incremental Growth
Dean, M. C. 1987. Growth layers and incremental markings in hard
tissues; a review of the literature and some preliminary ob-
servations about enamel structures in Paranthropus boisei.
Journal of Human Evolution 16, 157–172.
Laboratory of Postnatal Ontogeny, Koltzov Institute of Develop-
mental Biology, Russian Academy of Sciences 2000. Recording
Structures of Terrestrial Vertebrates. http://idbras.idb.ac.ru/
POSTNAT/regist.htm. Retrieved 9 January 2003.
Neville, A. C. 1967. Daily growth layers in animals and plants.
Biological Reviews of the Cambridge Philosophical Society 42,
421–441.
Peabody, F. E. 1961. Annual growth zones in living and fossil
vertebrates. Journal of Morphology 108, 11–62.
Incremental Growth in Teeth
Dean, M. C. 1995. The nature and periodicity of incremental lines
in primate dentine and their relationship to periradicular bands
in OH 16 (Homo habilis), pp. 36–46 in Moggi-Cecci, J. (ed.)
Aspects of Dental Biology – Palaeontology, Anthropology, and
Evolution. Florence: International Institute for the Study of Man.
[human dentine]
FitzGerald, C. M. 1998. Do enamel microstructures have regular
time dependency? Conclusions from the literature and a large-
scale study. Journal of Human Evolution 35, 371–386. [dentine
and enamel]
Gordon, B. C. 1984. Selected bibliography of dental annular studies
on various mammals. Zooarchaeological Research News,
Supplement 2, 1–24.
Gordon, B. C. 1992. Archaeological Seasonality Using Incremental
Structures in Teeth: An Annotated Bibliography. (Zooarchaeo-
logical Research News, Special Publication)
Grue, H. and Jensen, B. 1979. Review of the formation of incremental
lines in tooth cementum of terrestrial animals. Danish Review of
Game Biology 11(3), 3–48.
Hillson, Simon 1986. Teeth. Cambridge: Cambridge University
Press.
Lieberman, D. and Meadow, R. H. 1992. The biology of cementum
increments (with an archaeological application). Mammal Review
22, 57–77. [cementum references, p. 58]
O’Brien, Christopher 2001. A re-evaluation of dental increment
formation in East African mammals: implications for wildlife
biology and zooarchaeology. ArchaeoZoologia 11, 43–63.
Pike-Tay, A. 1995. Variability and synchrony of seasonal indicators
in dental cementum microstructure of the Kaminuriak Rangifer
population. Archeaofauna 4, 273–284.
Ramirez Rozzi, F. 1998. Enamel structure and development and its
application in hominid evolution and taxonomy. Journal of
Human Evolution 35, 321–330.
Ramirez Rozzi, F. (ed.) 1998. [special issue on primate dentition
microstructure]. Journal of Human Evolution 35 (4–5).
Scheffer, V. B. and Myrick, A. C. 1980. A review of studies to 1970
of growth layers in the teeth of marine mammals, pp. 51–63 in
Perrin, W. F. and Myrick, A. C. (eds), Age Determination of
Toothed Whales and Sirenians (Report of the International
Whaling Commission, Special Issue 3). Cambridge: International
Whaling Commission. [cited in Hillson 1986].
Shellis, R. P. 1999. Utilization of periodic markings in enamel to
obtain information on tooth growth. Journal of Human Evolution
35, 387–400. [primate enamel].
Stallibrass, S. 1982. The use of cement layers for absolute ageing of
mammalian teeth: A selective review of the literature,
with suggestions for further studies and alternative applications,
pp. 109–126 in Wilson, B., Grigson, C. and Payne, S. (eds),
Ageing and Sexing Animal Bones from Archaeological Sites
(BAR British Series 109). Oxford: British Archaeological
Reports.
Trunova, Y. E. and Klevezal, G. A. 1999. [Interspecific variation in
the records of winter hibernation in incisor dentine of rodents].
Zoologicheskii Zhurnal 78, 1455–1464.
Lower Vertebrates – Incremental Growth
Castanet, J., Francillon-Viellot, H., Meunier, F. J. and de Ricqles, A.
1993. Bone and individual aging, pp. 245–283 in Hall, B. K.
(ed.), Bone Growth. Boca Raton: CRC Press.
Fish – Incremental Growth
Das, M. 1994. Age determination and longevity in fishes. Geront-
ology 40, 70–96.
Menon, M. D. 1950 The use of bones, other than otoliths, in
determining the age and growth rate of fishes. Journal du Conseil
16, 311–340.
Amphibians – Incremental Growth
Castanet, J. and Smirina, E. 1991. Introduction to the skeleto-
chronological method in amphibians and reptiles. Annales des
Sciences Naturelles Zoologie et Biologie Animale 11(4) [1990],
191–196.
Smirina, E. M. 1994. Age determination and longevity in amphibians.
Gerontology V 40, 133–146.
Reptiles – Incremental Growth
Castanet, J. 1994. Age estimation and longevity in reptiles. Geront-
ology 40, 174–192.
Castanet, J. and Smirina, E. 1991. Introduction to the skeleto-
chronological method in amphibians and reptiles. Annales des
Sciences Naturelles Zoologie et Biologie Animale 11(4) [1990],

33Using Osteohistology for Ageing and Sexing
191–196.
Reid, R. E. H. 1990. Zonal “growth rings” in dinosaurs. Modern
Geology 15, 19–48.
Reid, R. E. H. 1997a. Histology of bones and teeth, pp. 329–339 in
Currie, K. A. and Padian, K. (eds), Encyclopedia of Dinosaurs.
San Diego: Academic. [dinosaurs]
Reid, R. E. H. 1997b. How dinosaurs grew, pp. 403–413 in Farlow,
J. O. and Brett-Surman, M. K. (eds) The Complete Dinosaur.
Bloomington: Indiana University Press.
Birds – Incremental Growth
Van Neer, W., Noyen, K., De Cupere, B. and Beuls, I. 2002b. On
the use of endosteal layers and medullary bone from domestic
fowl in archaeozoological studies. Journal of Archaeological
Science 29, 123–134. (pp. 125–128).
Mammals – Incremental Growth
Klevezal, G. A. 1996. Recording Structures of Mammals. Balkema
Publishers: Amsterdam. [comprehensive review of growth lines
in mammals, with an extensive bibliography; original Russian
edition: 1988]
Ageing with Osteons
Amprino, R. and Godina, G. 1947. La struttura delle ossa nei
vertebrati. Richerche comparative negli anfibi e negli amnioti
(Commentationes Pontificia Academia Scientiarum 11[9]).
Enlow, D. H. and Brown, S. O. 1956-58. A comparative histological
study of fossil and recent bone tissues. Texas Journal of Science
8, 405–443; 9, 186–214; 10, 187–230.
Ageing with Osteons – Humans
Robling, A. G. and Stout, S. 2000. Histomorphometry of human
cortical bone:
Applications to age estimation, pp. 187–208 and appendix in
Katzenberg, M. A. and Saunders, S. R. (eds), Biological Anthrop-
ology of the Human Skeleton. Wiley-Liss: New York.
Sex and Bone Histology
Spencer, M. and Uhler, K. (comps.) 1955. The Structure, Com-
position and Growth of Bone 1930–1953. Washington D. C.:
Armed Forces Medical Library, Reference Division. (pp. 75–82:
Hormonal influence – Sex hormones) [lightly annotated biblio-
graphy]
References and Selected Relevant Materials
N.B.: Many of these works (especially those by Castanet, Klevezal,
the Laboratory of Postnatal Ontogeny, O’Brien, Rosenberg and
Simmons, and Singh and Gunberg) list additional sources.
Alliston, T. and Derynck, R. 2002. Interfering with bone remodelling.
Nature 416, 886–887.
Amprino, R. 1947. La structure du tissu osseux envisagée comme
expression de différences dans la vitesse de l’accroissement.
Archives of Biology 58, 315–330.
Amprino, R. and Bairati, A. 1936. Processi di ricostuzione e di
riassorbimente nella sostanza compatta della ossa dell’uomo.
Zeitschrift für Zellforschung und mikroskopische Anatomie 24,
439–511.
Amprino, R. and Godina, G. 1947. La struttura delle ossa nei
vertebrati. Richerche comparative negli anfibi e negli amnioti
(Commentationes Pontificia Academia Scientiarum 11[9]).
Alexander, R. McN. 1975. The Chordates. Cambridge: Cambridge
University Press.
Arakelyan, M. S. and Danielyan, F. D. 2000. [Growth and age in
some parthenogenetic and bisexual species of rock lizards
(Lacerta) from Armenia]. Zoologicheskii Zhurnal 79(5), 585–
590.
Arthur, S. M., Cross, R. A., Paragi, T. F. and Krohn, W. B. 1992.
Precision and utility of cementum annuli for determining ages of
fishers. Wildlife Society Bulletin. 20(4), 402–405.
Balthazard, V. and Lebrun 1911. Les canaux de Havers aux
différents âges. Annales d’Hygiène Publique et de Médecine
Légale 15, 144–152. [cited in Castanet et al. 1993]
Batulevicius, D., Pauziene, N. and Pauza, D. H. 2001. Dental
incremental lines in some small species of the European
vespertilionid bats. Acta Theriologica 46, 33–42.
Beasley, M. J., Brown, W. A. B. and Legge, A. J. 1992. Incremental
banding in dental cementum: Methods of preparation of teeth
from archaeological sites and for modern comparative specimens.
International Journal of Osteoarchaeology 2, 37–50.
Bell, K., Loveridge, N., Reeve, J., Thomas, C., Feik, S. and Clement,
J. 2001a. Influence of age and gender on remodelling clusters
(super-osteons) in the cortex of human femoral shaft. 47th Annual
Meeting, Orthopaedic Research Society, February 25–28, 2001,
San Francisco, California. http://www.jbjs.org/ORS_2001/pdfs/
0234.pdf. Retrieved April 2002.
Bell, K. L., Loveridge, N., Reeve, J., Thomas, C. D. L., Feik, S. A.
and Clement, J. G. 2001b. Super-osteons (remodelling clusters)
in the cortex of the femoral shaft: influence of age and gender.
Anatomical Record 264, 378–386.
Bloom, W. and Fawcett, D. W. 1975. A Textbook of Histology.
Philadelphia, Pa.: Saunders.
Bourne, G. H. (ed.) 1972. The Biochemistry and Physiology of
Bone. Twelve volumes. London: Academic.
Bowden, D. M., Teets, C., Witkin, J. and Young, D. M. 1979. Long
bone calcification and morphology, pp. 335–347 in Bowden, D.
M. (ed.) Aging in Nonhuman Primates. New York: Van Nostrand
Reinhold.
Brandt, J. and Klemenz, A. 1998. Methoden der akustischen
Rastermikroskopie zur Untersuchung des Knochengewebes. 2.
Symposium “Quantitative Sonographie in Klinik und
Forschung“... 19–21. März 1998, Halle (Saale). http://
www.medizin.uni-halle.de:81/impb/quan98.htm. Retrieved April
2002.
Brockstedt, H., Kassem, M., Eriksen, E. F., Mosekilde, L. and
Melsen, F. 1993. Age- and sex-related changes in iliac cortical
bone mass and remodelling. Bone 14, 681–91.
Bryuzgin, V. 1939. A procedure for investigating age and growth in
Reptilia. Comptes Rendus (Doklady) de l’Académie des
Sciences de l’URSS 23, 403–405.
Bubenik, G. A. 1990. Neuroendorine regulation of the antler cycle,
pp. 265–97 in Bubenik, G. A. and Bubenik, A. B., Horns,
Pronghorns, and Antlers. New York: Springer-Verlag.
Buffrénil, V. de 1980. Données préliminaire: sur la structure des
marques de croissance squelettiques chez les crocodiliens actuels
et fossiles. Bulletin de la Sociéte Zoologique de France 105,
355–361.
Buffrénil, V. de 1982. Données préliminaire sur la présence de
lignes d’arrêt de croissance périostiques dans la mandibule du
marsouin commun, Phocoena phocoena (L), et leur utilisation
comme indicateur de l’âge. Canadian Journal of Zoology 60,
2557–2567.
Buffrénil, V. de and Buffetaut, E. 1981. Skeletal growth lines in an
Eocene crocodile skull from Wyoming as an indicator of onto-
genic age and paleoclimate conditions. Journal of Vertebrate
Paleontology 1, 57–66.
Buffrénil, V. de and Castanet, J. 2000. Age estimation by skeleto-
chronology in the Nile monitor (Varanus niloticus), a highly
exploited species. Journal of Herpetology 34, 414–424.
Burr, D. B. 1992. Estimated intercortical bone turnover in the femur
of growing macaques: Implications for their use as models in
skeletal pathology. Anatomical Record 232, 180–189.
Burr, D. B., Nishikawa, R. Y. and Van Gerven, D. P. 1989. Bone

34 K. Dammers
growth and remodelling in Cayo Santiago-derived Macaca
mulatta. Puerto Rican Health Science Journal 8, 191–196. [cited
in Burr 1992]
Camacho, N. P., Rinnerthaler, S., Paschalis, E. P., Mendelsohn, R.,
Boskey, A. L. and Fratzel, P. 1999. Complementary information
on bone ultastructure from scaling small angle X-ray scattering
and Fourier-transform infrared microscopy. Bone 25, 287–293.
Caplan, A. I. 1998. Bone development, pp. 3–21 in Evered, D. and
Harnett, S. (eds) Cell and Molecular Biology of Vertebrate
Hard Tissues. Chichester: Wiley.
Carlson, S. 1990. Vertebrate dental structures, pp. 531–56 in Carter,
J. G. (ed.) Skeletal Biomineralization Patterns, Processes and
Evolutionary Trends, Volume I. New York: Van Nostrand
Reinhold. [See especially pp. 542–544: “Incremental Lines and
Growth Bands in Vertebrate Dental Tissues.”]
Carter, J. G. (comp.) 1990. Glossary of skeletal biomineralization,
pp. 609–671 in Carter, J. G. (ed.), Skeletal Biomineralization:
Patterns, Processes and Evolutionary Trends, Volume I. New
York: Van Nostrand Reinhold.
Castanet, J. 1974. Etude histologique des marques squelettiques de
croissance chez Vipera aspis (L.) (ophidia, viperidae). Zoologica
Scripta 3, 137–151.
Castanet, J. 1975. Quelques observations sur la présence et la
structure des marques squelettiques de croissance chez les
amphibiens. Bulletin de la Sociéte Zoologique de France 100,
603–631.
Castanet, J. 1978. Les marques de croissance osseuse comme
indicateurs de l’âge chez les lézards. Acta Zoologica 59, 35–48.
Castanet, J. and Baez, M. 1991. Adaptation and evolution in Gallotia
galloti lizards from the Canary Islands: Age, growth, maturity
and longevity. Amphibia-Reptilia 12, 81–102.
Castanet, J. and Cheylan, M. 1979. Les marques de croissance des
os et des écailles comme indicateur de l’âge chez Testudo
hermanni et Testudo graeca (Reptilia, Chelonia, Testudinidae).
Canadian Journal of Zoology 57, 1649–1665.
Castanet, J. and Gasc, J. P. 1986. Age individuel longèvité et cycle
d’activité chez Leposoma guianense microteiidé de litière de
l’écosystème forestier guyianais. Mémoire Muséum National
d’Histoire Naturelle (Paris) 132, 281–288.
Castanet, J. and Roche, E. 1981. Determination de l’age chez le
lezard des murailles, Lacerta muralis (Laurenti, 1768) au moyen
de la squelettochronologie, Revue Suisse de Zoologie 88, 215–
226.
Castanet, J. and Smirina, E. 1991. Introduction to the skeleto-
chronological method in amphibians and reptiles. Annales des
Sciences Naturelles Zoologie et Biologie Animale
11(4)[1990],191–196.
Cattaneo, C., DiMartino, S., Scali, S., Craig, O. E., Grandi, M. and
Sokol, R. J. 1999. Determining the human origin of fragments of
burnt bone: A comparative study of histological, immunological
and DNA techniques. Forensic Science International 102, 181–
191.
Cerroni A. M., Tomlinson, G. A., Turnquist, J. E. and Grynpas, M.
D. 2000. Bone mineral density, osteopenia, and osteoporosis in
the rhesus macaques of Cayo Santiago. American Journal of
Physical Anthropology 113, 389–410.
Chamberlain, A. and Forbes, S. 2001. A preliminary study of
microscopic evidence for lactation in cattle. Unpublished paper
delivered at the Osteon Workshop, Department of Physical
Anthropology, University of Göttingen, September 2001.
Chamberlain, A.T. and Forbes, S.T. (in press) A preliminary study
of microscopic evidence for lactation in cattle, in Mulville, J.
(ed.) Proceedings of the International Council for Archaeo-
zoology.
Champ, J. E., Binkley, N., Havighurst, T., Colman, R. J., Kemnitz,
J. W. and Roecker, E. B. 1996. The effect of advancing age on
bone mineral content of female rhesus monkeys. Bone 19, 485–
492.
Chaplin, R. E. 1971. The Study of Animal Bones from Archaeo-
logical Sites. London: Seminar.
Chinsamy, A. 1993. Bone histology and growth trajectory of the
prosauropod dinosaur Massospondylus Carinatus Owen,
Modern Geology 18, 319–329.
Chinsamy, A. 1995. Ontogenetic changes in the bone histology of
the Late Jurassic ornithopod Dryosaurus lettowvorbecki. Journal
of Vertebrate Paleontology 15, 96–104.
Chinsamy, A. and Elzanowski, A. 2001. Bone histology: Evolution
of growth pattern in birds. Nature 412, 402–403.
Chinsamy, A., Chiappe, L. M. and Dodson, P. 1994. Growth rings
in Mesozoic birds. Nature 368, 196–197.
Chinsamy, A. and Raath, M. A. 1992. Preparation of fossil bone for
histological examination. Palaeontologia Africana 29, 39–44.
Colman, R. J., Kemnitz, J. W., Lane, M. A., Abbott, D. H. and
Binkley, N. 1999. Skeletal effects of aging and menopausal status
in female rhesus macaques. Journal of Clinical Endochrinology
and Metabolism 84, 4144–4148.
Condon, K., Charles, D. K., Cheverud, J. M. and Buikstra, J. E.
1986. Cementum annulation and age determination in Homo
Sapiens. II Estimates and accuracy. American Journal of
Physical Anthropology 71, 321–330.
Constans, Aileen 2001. Tools and technology: Pretty on the inside.
The Scientist 15 (12), 25.
Cool, S. M., Forwood, M. R., Campbell, P. and Bennett. M. B.
2002. Comparisons between bone and cementum compositions
and the possible basis for their layered appearances. Bone 30,
386–392.
Coy, J. P., Jones, R. T. and Turner, K. A. 1982. Absolute ageing of
cattle from tooth sections and its relevance to archaeology, pp.
127–40 in Wilson, B., Grigson, C. and Payne, S. (eds), Ageing
and Sexing Animal Bones from Archaeological Sites (BAR
British Series 109). Oxford: British Archaeological Reports.
Cuezva, S. and Elez I., J. 2000. Rconocimiento del estadio de
desarrollo en la microestructura de los huesos fósiles de
mamíferos (somosaguas y Layna). Coloquios de Paleontología
51, 159–174.
Curry Rogers, K. 2000. Growth rates among the dinosaurs, pp.
297–309 in Paul, G. S. (ed.), The Scientific American Book of
Dinosaurs. New York: St. Martin’s (in press).
Das, M. 1994. Age determination and longevity in fishes. Geront-
ology 40, 70–96.
Davis, S. J. M. 1987. The Archaeology of Animals. London:
Batsford.
Demeter, G. and. Mátyás, J. 1928. Mikroskopisch vergleichend-
Rücksicht auf die Unterscheidung menschlicher und tierischer
Knochen. Zeitschrift für Anatomie und Entwicklungsgeschichte
87, 45–99.
Drew, I. M., Perkins Jr., D. and Daly, P. 1971. Prehistoric
domestication of animals: effects on bone structure. Science 171,
280–282.
Driver, J. C. 1982. Medullary bone as an indicator of sex in bird
remains form archaeological sites, pp. 251–4 in Wilson, B.,
Grigson, C. and Payne, S. (eds), Ageing and Sexing Animal
Bones from Archaeological Sites (BAR British Series 109).
Oxford: British Archaeological Reports.
Dubick, J. D. 2000. Age and growth of the spotted eagle ray,
Aetobatus narinari (Euphrasen, 1790) from southwest Puerto
Rico with notes on its biology and life history. Unpublished M.S.
thesis, University of Puerto Rico. [abstract]
Ducy, P., Schinke, T. and Karsenty, G. 2000. The osteoblast: A
sophisticated fibroblast under central surveillance. Science 289,
1501–1504.
Eckardt, I. and Hein, H.J. 1999. Quantitative measurements of the
mechanical properties of human bone tissues by SAM.
Augustusburg Conference of Advanced Science: Mechanical

35Using Osteohistology for Ageing and Sexing
Properties of Cells and Tissues. 11–13 October 1999,
Augustusburg, Germany. http://www.medizin.uni-alle.de:
81/biomechanik/acas99/abstracts/eckardt.htm. Retrieved April
2002.
Egeli, R. 1976. Verhalten der Osteongroßen in der Knochen-
kompakta des Menschen in Abhängigkeit von Alter und
Geschlecht. Unpublished Diss.med., Bern University. Fifteen
pages. [listed in Bibliographie Medizin 1993, Nr. 1114666]
El Mounden, E., Francillon-Viellot, H., Castanet, J. and Znari, M.
1997. A skeletochronological study of age, maturity, growth and
longevity in the North African agamind, Agama impalearis
Boettger, 1874. Annales des Sciences Naturelles Zoologie et
Biologie Animale 18, 63–70.
Enlow, D. H. 1963. Principles of Bone Remodelling. Springfield:
Thomas.
Enlow, D. H. 1969. The bone of reptiles, pp. 45–80 in Gans, C.
(ed.), Biology of the Reptilia, vol. 1. Morphology A. New York:
Academic.
Enlow, D. H. and Brown, S. O. 1956–58. A comparative histological
study of fossil and recent bone tissues. Texas Journal of Science
8, 405–443; 9, 186–214; 10, 187–230.
Erickson, G. M. 1997. Age determination of dinosaurs, pp. 4–6 in
Currie, P. J. and Padian, K. (eds), Encyclopedia of Dinosaurs.
San Diego: Academic Press.
Erickson, G. M., Curry Rogers, K., Yerby, S. A. 2001. Dinosaurian
growth patterns and rapid avian growth rates. Nature 412, 429–
433.
Erickson, G. M. and Tumanova, T. 1995. Histological variation
through ontogeny in the long bones of Psittacosaurus mongo-
liensis. Journal of Vertebrate Paleontology 15 (Supplement to
Number 3), 28A.
Erickson, G. M. and Tumanova, T. A. 2000. Growth curve and life
history attributes of Psittacosaurus mongoliensis (Ceratopsia:
Psittacosauridae) inferred from long bone histology. Zoological
Journal of the Linnean Society 130, 551–566.
Erişmiş, U. C., Kaya, U. and Arikan, H. 2002. Observations on the
histomorphological structure of some long bones of the water
frog (Rana bedriagae) from the İzmir area. Turkish Journal of
Zoology 26, 213–216.
Esteban, M., Garcia Paris, M. and Castanet, J. 1996. Use of bone
histology in estimating the age of frogs (Rana perezi) from a
warm temperate climate area. Canadian Journal of Zoology 74,
1914–1921.
Fajardo, R. J., Ryan, T. M. and Kappelman, J. 2002. Assessing the
accuracy of high-resolution X-ray computed tomography of
primate trabecular bone by comparisons with histological
sections. American Journal of Physical Anthropology 118, 1–
10.
Ferguson, M. W. J., Honig, L. S., Bringas Jr., P. and Slavkin, H. C.
1982. In vivo and in vitro development of first branchial arch
derivatives in Alligator mississippiensis, pp. 275–86 in Dixon,
A. D. and Sarnat, B. (eds), Factors and Mechanisms Influencing
Bone Growth. New York: Alan R. Liss.
Filogamo, G. 1946. La forme et la taille des ostéones chez quelques
mammifères. Archives de Biologie 57, 137–143.
FitzGerald, C. M. 1998. Do enamel microstructures have regular
time dependency? Conclusions from the literature and a large-
scale study. Journal of Human Evolution 35, 371–386.
Foote, J. S. 1913. The comparative histology of the femur.
(Smithsonian Miscellaneous Collections, vol. 61, No. 8).
Washington, D.C.
Forbes, S. T. 1994. Palaeohistology: The potential of this technique
for investigating the advent of domestication and the secondary
products revolution. Unpublished M.Sc. thesis, Department of
Archaeology and Prehistory, University of Sheffield. [cited in
Chamberlain and Forbes 2001]
Fox, D. L. 2000. Growth increments in Gomphotherium tusks and
implications for late Miocene climate change in North America.
Palaeogeography, Palaeoclimatology, Palaeoecology 156, 327–
348.
Francillon-Viellot, H., Buffrénil, V. de, Castanet, J., Géraudie, J.,
Meunier, F. J., Sire, J. Y., Zylberg, L. and Ricqles, A. de 1990.
Microstructure and mineralization of vertebrate skeletal tissues,
pp. 471–530 in Carter J. G. (ed.), Skeletal Biomineralization of
Patterns, Processes and Evolutionary Trends. Two volumes.
New York: Van Nostrand Reinhold.
Frost, H. M. 1964. Mathematical Elements of Lamellar Bone
Remodelling. Thomas: Springfield.
Garcia-Perea, R. and Baquero, R. A. 1999. Age estimation in Iberian
wildcats Felis silvestris, by canine tooth sections. Acta
Theriologica 44, 321–327.
Gebhardt, W. 1906. Über funktionell wichtige Anordnungsweisen
der feineren und gröberen Bauelemente des Wirbeltierknochens.
II. spezieller Teil. 1. Der Bau der Haverschen Lamellensysteme
und seine funktionelle Bedeutung. W. Roux Archiv für
Entwicklungsmechanic der Organismen 20, 187–334.
Gilbert, A. S. 1989. Microscopic bone structure in wild and domestic
animals: A reappraisal, pp. 47–86 in Crabtree, P. J., Campana,
D. and Ryan, K. (eds), Early Animal Domestication and Its
Cultural Context. (MASCA Research Papers in Science and
archaeology, Volume 6 Supplement). Philadelphia: University
of Pennsylvania.
Glorieux, F. H., Travers, R., Taylor, A., Bowen, J. R., Rauch, F.,
Norman, M. and Parfitt, A. M. 2000. Normative data for iliac
bone histomorphometry in growing children. Bone 26, 103–109.
Gordon, B. C. 1993. Archaeological tooth and bone seasonal
increments: The need for standardized terms and techniques.
ArchaeoZoologia 5, 9–16.
Griffith, I. 1962. Skeletal lamellae as an index of age in hetero-
themous tetrapods. Annals and Magazine of Natural History,
Zoology, and Botany, Series 13, 449–465.
Gross, W. 1934. Die Typen des mikroskopischen Knochenbaues bei
fossilen Stegocephalen und Reptilien. Zeitschrift für Anatomie
und Entwicklungsgeschichte 103, 731–764. [cited in Reid 1990]
Grue, H. and Jensen, B. 1979. Review of the formation of incremental
lines in tooth cementum of terrestrial animals. Danish Review of
Game Biology 11(3), 3–48.
Grupe, G. and Herrmann, B. 1983/84. Über das Schrumpfverhalten
experimentell verbrannter Knochen am Beispiel des Caput
femoris. Zeitschrift für Morphologie und Anthropologie 74,
121–127.
Grynpas, M. D., Huckell, B., Pritzker, K. P. H., Hancock, R. G. V.
and Kessler, M. J. 1989. Bone mineral and osteoporosis in aging
rhesus monkeys. Puerto Rico Health Sciences Journal 8, 197–
204. [cited in Lees and Ramsay 1999]
Gustafson, G. 1950. Age determination of teeth. Journal of the
American Dental Association 41, 45–54.
Guyetant, R., Castanet, J. and Pinston, H. 1984. Determination de
l’age de jeunes grenouilles, Rana temporaria L., par l’analyse
des marques de croissance de coupes transversales d’os compact.
Comptes Rendus des Seances de la Societe de Biologie et de
Ses Filiales 178, 271–777.
Harsányi, L. 1993. Differential diagnosis of human and animal bone,
pp. 79–94 , pp. 111–123 in Grupe, G. and Garland, A. N. (eds),
Histology of Ancient Human Bone: Methods and Diagnosis.
Berlin: Springer.
Havers, C. 1691. Osteologia Nova or Some New Observations of
the Bones. London: Samuel Smith.
Havill, L. M. 2002. Osteon remodelling dynamics in the rhesus
macaque: Normal variation and the genetic contribution.
Unpublished Ph.D. thesis, Dept. of Anthropology, Indiana
University, Bloomington.
Havill, L. M. 2004. Osteon remodelling dynamics in Macaca mulatta:
Normal variation with regard to age, sex, and skeletal maturity.

36 K. Dammers
Calcified Tissue International 74 (1), 95–102.
Henderson, B. A. and Bowen, H. M. 1979. A short note. Journal of
Applied Ecology 16, 363–366.
Henning, S., Adhikari, R. Godehardt, R., Michler, G. H., Matern, D.
and Hein, H. J. 1999. Atomic force microscopic analysis of size,
shape and organization of mineral crystals in human cortical
bone. Augustusburg Conference of Advanced Science:
Mechanical Properties of Cells and Tissues. 11–13 October
1999, Augustburg, Germany.
http://www.medizin.uni-halle.de:81/biomechanik/acas99/
abstracts/henning.htm. Retrieved April 2002.
Hidaka, S., Matsumoto, M. Ohsako, S., Toyoshima, Y. and
Nishinakagawa, H. 1998. A histometrical study on the long bones
of raccoon dogs, Nyctereutes procyonoides and badgers, Meles
meles. Journal of Veterinary Medical Science 60, 323–326.
Hillson, S. 1986. Teeth. Cambridge Manuals in Archaeology.
Cambridge: Cambridge University Press.
Hittel, T. H. 1860. The Adventures of James Capen Adams,
Mountaineer and Grizzly Bear Hunter, of California. San
Francisco: Towne & Bacon. [cited in Peabody 1961; see p. 291.]
Hodges, R. D. 1974. The Histology of Fowl. London: Academic
Press.
Horner, J. R., de Ricqles, A. and Padian, K. 1999. Variation in
dinosaur skeletochronology indicators: Implications for age
assessment and physiology. Paleobiology 25, 295–304.
Horner, J. R., de Ricqles, A. and Padian, K. 2000. Long bone
histology of the hadrosaurid dinosaur Maiasaura peeblesorum:
Growth dynamics and physiology based on an ontogenetic series
of skeletal elements. Journal of Vertebrate Paleontology 20,
115–129.
Hummel, S. 2001. Die Bearbeitung von Menschlichen
Leichenbränden (Blockkurs). Skript zum Praktikum. Seminar
der Ur- und Frühgeschichte Universität Basel.
Hummel, S. and Schutkowski, H. 1986. Das Verhalten von
Knochengewebe unter dem Einfluß höherer Temperaturen.
Bedeutung für die Leichenbranddiagnose. Zeitschrift für Morph-
ologie und Anthropologie 77, 1–9.
Hummel, S. and Schutkowski, H. 1993. Approaches to the
histological age determination of cremated human remains, pp.
111–123 in Grupe, G. and Garland, A. N. (eds), Histology
of Ancient Human Bone: Methods and Diagnosis. Berlin:
Springer.
Hutton, J. M. 1986. Age determination of living Nile crocodiles
from the cortical stratification of bone. Copeia (2), 332–341.
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 Osteo-
archaeology 11, 415–432.
Jones, S. J., Glorieux, F. H., Travers, R. and Boyde, A. 1988. The
microscopic structure of bone in normal children and patients
with osteogenesis imperfecta: A survey using backscattered
electron imaging. Calcified Tissue International 64, 8–17.
Kagerer, P. and Grupe, G. 2001. Age-at-death diagnosis and
determination of life-history parameters by incremental lines in
human dental cementum as an identification aid. Forensic
Science International 118, 75–82.
Kardong, K. V. 1995. Vertebrates: Comparative anatomy, function,
evolution. Dubuque, Iowa: Wm. C. Brown.
Katz, J. L. and Bumrerraj, S. 1999. Scanning acoustic microscopy
studies of cortical and trabecular bone. Augustusburg Conference
of Advanced Science: Mechanical Properties of Cells and Tissues.
11–13 October 1999, Augustburg, Germany.
http://www.medizin.uni-halle.de:81/biomechanik/acas99/
abstracts/katz.htm. Retrieved April 2002.
Katz, J. L. and Meunier, A. 1993. Scanning acoustic microscope
studies of the elastic properties of osteons and osteon lamellae.
Journal of Biomechanical Engineering 115 (4, Part B), 543–
548.
Kay, R. F., Rasmussen, D. T. and Beard K. C. 1984. Cementum
annulus counts provide a means for age determination in Macaca
mulatta (Primates. Anthropoidea). Folia Primatologica 42, 85–
95.
Kenyeres, B. and Hegyi, M. 1903. Unterscheidung des menschlichen
und des thierischen Knochengewebes. Vierteljahresschrift für
gerichtliche Medicin und öffentliches Sanitätswesen 3. Folge
XXV, 2, 22–32.
Kerley, E. R. 1965. The microscopic determination of age in human
bone. American Journal of Physical Anthropology 23, 149–164.
Kierdorf, U., Kierdorf, H. and Knuth, S. 1995. Effects of castration
on antler growth in fallow deer (Dama dama L.). Journal of
Experimental Zoology 273, 33–43.
King, C. M. 1991. A review of age determination methods for the
stoat Mustela erminea. Mammal Review 21, 31–49.
Kingsley, D. M. 2001. Genetic control of bone and joint formation,
pp. 213-234 in Cardew, G. and Goode, J. A. (eds), The Molecular
Basis of Skeletogenesis. Chichester: Wiley.
Klebanova, E. A. and Klevezal, G. A. 1966. [Stratification of the
periosteal zone of tubular bones in limbs as a criterion for age
determination in mammals]. Zoologicheskii Zhurnal 45, 406–
413.
Kleinenberg, S. E. and Smirina, E. M. 1969. [A contribution to the
method of age determination in amphibians]. Zoologicheskii
Zhurnal 48, 1090–1094.
Klevezal, G. A. 1996. Recording Structures of Mammals. Balkema
Publishers: Amsterdam. [original Russian edition: 1988]
Klevezal, G. A. 2001. [Assessment of life history and status at the
death of Apodemus mice from recording structures].
Zoologicheskii Zhurnal 80, 599–606. [abstract]
Klevezal, G. A. 2002. Reconstruction of individual life histories of
rodents from their teeth and bone. Acta Theriologica 47 (Supple-
ment 1), 127–138.
Klevezal, G. A. and Kleinenberg, S. E. 1969. Age Determination of
Mammals from Annual Layers in Teeth and Bones. Academy of
Sciences U.S.S.R. Israel Program for Scientific Translation:
Jerusalem.
Klevezal, G. A. and Sukhovskaya, L. I. 1995. [Dentine of incisors of
rodents as a recording structure]. Zoologicheskii Zhurnal 74
(4), 124–131.
Korkmaz, T., Kaya, M. A. and Bayazit, V. 1996. Relation with the
age of the histological variations in the compact layer of the long
bones. Turkish Journal of Zoology 20, 71–76.
Kumbar, S. M. and Pancharatna, K. 2001. Determination of age,
longevity and age at reproduction of the frog Microhyla ornata
by skeletochronology. Journal of Biosciences 26, 265–270.
Lapeña, M., Patón, D., Hernández, F., de Lope, F. and Juarranz, A.
1993. Two examples showing contradictory results by using
skeletochronology in birds. Archaeofauna 2, 175–9. [abstract
retrieved 3 January 2003: http://www.uam.es/otros/paleofau/
Revista/Vol2/volumen2twoexamples.htm]
Lasota-Moskalewska, A. and Moskalewski, S. 1982. Microscopic
comparison of bones from Medieval domestic and wild pigs.
Ossa 7, 173–178.
Laws, R. M. 1952. A new method of age determination for mammals.
Nature 169, 972–973.
Laws, R. M. 1953. The elephant seal (Mirounga leonina Linn.):
growth and age. Falkland Islands Dependencies Survey,
Scientific Report 8, 1–62.
Lees, C. J. and Ramsay, H. 1999. Histomorphometry and bone
biomarkers in cynomolgus monkeys: Astudy in young, mature,
and old monkeys. Bone 24, 25–28.
Lieberman, D. and Meadow, R. H. 1992. The biology of cementum
increments (with an archaeological application). Mammal Review
22, 57–77.
Lima, V., Arntzen, J. W. and Ferrand, N. M. 2001. Age structure

37Using Osteohistology for Ageing and Sexing
and growth pattern in two populations of the golden-striped
salamander Chioglossa lusitanica (Caudata, Salamandridae),”
Amphibia-Reptilia 22, 55–68. [abstract]
Lind, R. 1994. Health monitoring bones. http:/ase.aero.ufl.edu/rick/
rick_pro/rick_bones.html. Retrieved 13 January 2003.
Lynnerup, N., Thomsen, J. L. and Frohlich, B. 1998. Intra- and
inter-observer variation in histological criteria used in age at
death determination based on femoral cortical bone. Forensic
Science International 91, 219–230.
Maat, G. J. R., Van Den Bos, R. P. M. and Aarents, M. 2001.
Manual preparation of ground sections for the microscopy of
natural bone tissue: Update and modification of Frost’s ‘rapid
manual method,’ International Journal of Osteoarchaeology
11, 366–374.
Maples, W. R. and Rice, P. M. 1979. An improved technique using
dental histology for estimation of adult age. Forensic Science
International 23, 764–770.
Martin, D. and Armelagos, G. J. 1979. Morphometrics of compact
bone: An example from Sudanese Nubia. American Journal of
Physical Anthropology 51, 571–579.
Martin, R. B., Burr, D. B. and Sharkey, N. A. 1998. Skeletal Tissue
Mechanics. New York: Springer.
Mattox, N. T. 1936. Annular rings in the long bones of turtles and
their correlation with size. Transactions of the Illinois State
Academy of Science 28, 255–256.
Mays, S. 1998. The Archaeology of Human Bones. London:
Routledge.
Mátyás, J. 1927. Die mikroanatomische Knochenstruktur und die
Abstammungsgeschichte einzelner Wirbeltiergruppen. Xe
Congrès International de Zoologie, 606–704 and Plates I–XLII.
McKinley, V. and Burke, A. 2001. A new control sample for season
of death estimates for Equus caballus from dental thin-sections.
ArchaeoZoologia 11, 33–42.
Meindl, R. S. and Russell, K. F. 1998. Recent advances in method
and theory in paleodemography. Annual Review of Anthropology
27, 375–399.
Miani, A. and Miana, C. 1972. Circadian advancement rhythm of
the calcification front in dog dentin. Panminerva Medica 14,
127–136 [cited in Rosenberg and Simmons 1980].
Miller, F. L. 1974. Age determination of caribou by annulations in
dental cementum. Journal of Wildlife Management 38, 47–53.
Moskilde, L. 2001. Mechanisms of age-related bone loss, pp. 150–
171 in Bock, G. and Gode, J. A. (eds), Ageing Vulnerability.
New York: Wiley.
Mulhern, D. M. and Ubelaker, D. H. 2003. Histologic examination
of bone development in juvenile chimpanzees. American Journal
of Physical Anthropology 122, 127–133.
Nopsca, F. B. von and Heidsieck, E. 1933. On the histology of the
ribs in immature and half-grown trachodont dinosaurs. Pro-
ceedings of the Zoological Society of London 1, 221–226.
Nouira, S., Maury, M. E., Castanet, J. and Barbault, R. 1982.
Determination squelettochronologique de l’age dans une popu-
lation de Cophosaurus texanus (Sauria, Iguanidae). Amphibia-
Reptilia 3, 213–219.
O’Brien, Christopher 2001. A re-evaluation of dental increment
formation in East African mammals: Implications for wildlife
biology and zooarchaeology. ArchaeoZoologia 11, 43–63.
O’Connor, T. 2000. The Archaeology of Animal Bones. Stroud,
Gloucestershire: Sutton.
Ohtsuka-Isoza, M., Hayashi, H. and Shinoda, H. 2001. Effect of
suprachiasmatic nucleus lesion on circadian dentin increment in
rats. American Journal of Physiology 280 (5), 1364–1370.
Okada, M. and Mimura, T. 1941. Zur Physiologie und Pharmak-
ologie der Hartegewebe. VII. Über den zeitlichen Verlauf der
Schwangerschaft und Entbindung gesehen von der Streifenfigur
im Dentin des mütterliches Kaninchens. Proceedings of the
Japanese Pharmacological Society 15th Meeting, Japanese
Journal of Medical Sciences IV. Pharmacology 14, 7–10. [cited
in Rosenberg and Simmons 1980]
Okada, M. 1943. Studies in the periodic patterns of hard tissues in
animal body. Shanghai Evening Post, Medical Edition,
September, pp. 26–31. [cited in Rosenberg and Simmons 1980]
Ohtaishi, N. and Hachiya, N. 1985. Ageing techniques from annual
layers in teeth and bone, pp. 186–190 in Contemporary Mam-
malogy in China and Japan. Mammalogical Society of Japan.
[listed in the Zoological Record]
Ott, S. M., Lipkin, E. W. and Newell-Morris, L. 1999. Bone
physiology during pregnancy and lactation on young macaques.
Journal of Bone and Mineral Research 14, 1779–1788.
Padian, K. 1997. Growth lines, pp. 288–291 in Currie, P. J. and
Padian, K. (eds), Encyclopedia of Dinosaurs. San Diego:
Academic.
Parfitt, A. M. 1993. Morphometry of bone resorption: Introduction
and overview. Bone 14, 435–441.
Parfitt, A. M., Drezner, M. K., Glorieux, F. H., Kans, J. A., Malluche,
H., Meunier, P. J., Ott, S. M. and Recker, R. R. (ASBMR
Nomenclature Committee) 1987. Bone histomorphometry:
Standardization of nomenclature. Journal Bone and Mineral
Research 2, 595–610.
Pascal, M. and Castanet, J. 1978. Méthode de détermination de
l’âge chez le chat haret des îles Kerguelen. La Terre et la Vie 32,
529–555.
Patnaik, B. K. and Behera, M. N. 1981. Age determination in the
tropical agamid lizard, Calotes versicolor (Daudin) based on
bone histology. Experimental Gerontology 16, 295–308.
Pattijn, V., Van Oosterwyck, H., Vander Sloten, J., Van der Perre,
G., Naert, I., Wevers, M. and Jansen, J. 1999. The use of
microfocus computer tomography for the non-destructive
characterisation of trabecular bone structures. Augustusburg
Conference of Advanced Science: Mechanical Properties of
Cells and Tissues. 11–13 October 1999, Augustburg, Germany.
http://www.medizin.uni-halle.de:81/biomechanik/acas99/
abstracts/pattijn.htm. Retrieved April 2002.
Pavao, B. and Stahl, P. W. 1999. Structural density assay of leporid
skeletal elements with implications for taphonomic, actualistic,
and archaeological research. Journal of Archaeological Science
26, 53–66.
Peabody, F. E. 1958. A Kansas drought recorded in growth zones of
a bullsnake. Copeia (2), 91–94.
Peabody, F. E. 1961. Annual growth zones in living and fossil
vertebrates. Journal of Morphology 108, 11–62.
Petter-Rousseau, A. 1953. Recherches sur la croissance et le cycle
d’activité testiculaire de Natrix natrix helvetica (Lacépède). La
Terre et la Vie 4, 175–223.
Pfeiffer, S. 1992. Cortical bone age estimates from historically known
adults. Zeitschrift für Morphologie und Anthropologie 79, 1–
10.
Pfeiffer, S. 2000. Palaeohistology: Health and disease, pp. 287-302
in Katzenberg, M. A. and Saunders, S. R. (eds), Biological
Anthropology of the Human Skeleton. Wiley-Liss: New York.
Pike-Tay, A. 1995. Variability and synchrony of seasonal indicators
in dental cementum microstructure of the Kaminuriak Rangifer
population. Archeaofauna 4, 273–284.
Pike-Tay, A., Valdes, V. C. and Quiros, F. B. de 1999. Seasonal
variation of the Middle-Upper Paleolithic transition at El Castillo,
Cueva Morin and El Pendo (Cantabria, Spain). Journal of Human
Evolution 36, 283–317.
Poulsen, L. W., Qvesel, D., Brixen, K., Vesterby, A. and Boldsen, J.
L. 2001. Low bone mineral density in the femural neck of
medieval women. Bone 28, 454–458.
Pritchard, J. J. 1972. General histology of bone, pp. 1–20 in Bourne,
G. H. (ed.) The Biochemistry and Physiology of Bone, vol. 1.
Twelve volumes. London: Academic.
Przybeck, T. R. 1985. Histomorphology of the rib: Bone mass and

38 K. Dammers
cortical remodelling, pp. 303–326 in Davis, R. T. and
Leathers, C. W. (eds) Behavior and pathology of aging in rhesus
monkeys. New York: Alan R. Liss.
Quekett, J. 1849. On the different types in the microscopic structure
of the skeleton in the four great classes of animals, viz. mammals,
birds, reptiles and fishes. Transactions of the Microscopical
Society of London 2, 40–42.
Quekett, J. 1855. Descriptive and Illustrated Catalogue of the
Histological Series Contained in the Museum of the Royal
College of Surgeons. II. Structure of the Skeleton of Vertebrate
Animals. London: Royal College of Surgeons of London.
Ray, S. and Chinsamy, A. 2004. Diictodon feliceps (Therapsida,
Dicynodontia): Bone histology, growth, and biomechanics.
Journal of Vertebrate Paleontology 24, 180–194.
Rehman, M. T. A., Hoyland, J. A., Denton, J. and Freemont, A. J.
1994. Age related histomorphometric changes in bone in normal
British men and women. Journal of Clinical Pathology 47, 529–
534.
Reid, R. E. H. 1990. Zonal “growth rings” in dinosaurs. Modern
Geology 15, 19–48.
Reid, R. E. H. 1997. Histology of bones and teeth, pp. 329–339 in
Currie, K. A. and Padian, K. (eds), Encyclopedia of Dinosaurs.
San Diego: Academic.
Rensberger, J. M. and Watabe, M. 2000. Fine structure of bone in
dinosaurs, birds and mammals. Nature 406, 619–621.
Report of the Workshop 1980. pp. 1–50 in Perrin, W. F. and Myrick,
A. C. (eds), Age Determination of Toothed Whales and Sirenians
(Report of the International Whaling Commission, Special Issue
3). Cambridge: International Whaling Commission. [cited in
Hillson 1985]
Richardson, C. R., Fisher, A. K. and Folk, G. E. 1961. The dental
tissues of wild and laboratory-raised hibernating and non-
hibernating 13-lined ground squirrels. Journal of Dental Re-
search 40, 1029–1035.
Robling, A. G. and Stout, S. 2000. Histomorphometry of human
cortical bone: Applications to age estimation, pp. 187–208 and
appendix in Katzenberg, M. A. and Saunders, S. R. (eds),
Biological Anthropology of the Human Skeleton. Wiley-Liss:
New York.
Robling, A. G. and Stout, S. D. 1999. Morphology of the drifting
osteon. Cells Tissues Organs 164 (4), 192–204.
Rosenberg, G. D. and Simmons, D. J. 1980. Rhythmic dentinogenesis
in the rabbit incisors: Circadian, ultradian, and infradian periods.
Calcified Tissue International 32, 29–44.
Ruddle, J. L. 1997. An investigation of bone histology as a potential
age indicator in roe deer. Unpublished Ph.D. thesis, University
College, London. [abstract, viewable at http://www.theses.com/
idx/048/it048007816.htm]
Rudge, M. R. 1976. Aging domestic sheep (Ovis aries L) from
growth lines in cementum of first incisor. New Zealand Journal
of Zoology 3, 421–424.
Russell, K. F. 1996. Determination of age-at-death from dental
remains. Unpublished Ph.D. thesis, Kent State University.
[abstract: Dissertation Abstracts International 58 (2), 502–3–
A]
Saint-Girons, H. 1965. Les critères d’âge chez les reptiles et leurs
applications à l’etude de la structure des populations sauvages.
La Terre et la Vie 19, 342–358.
Schaffler, M. B. and Burr, D. B. 1984. Primate cortical bone
microstructure: Relationship to locomotion. American Journal
of Physical Anthropology 65, 191–197.
Scheffer, V. B. 1950. Growth layers on the teeth of pinnipedia as
indicators of age. Science 112, 309–311.
Scheffer, V. B. and Myrick, A. C. 1980. A review of studies to 1970
of growth layers in the teeth of marine mammals, pp. 51–63 in
Perrin, W. F. and Myrick, A. C. (eds), Age Determination of
Toothed Whales and Sirenians (Report of the International
Whaling Commission, Special Issue 3). Cambridge: International
Whaling Commission. [cited in Hillson 1986]
Schmidt, U. J., Kalbe, I. and Mühlbach, R. 1981. Alterns-
veränderungen des Knochengewebes. Gegenbaurs Morpho-
logisches Jahrbuch 127, 636–640.
Schour, I. and Steadman, S. R. 1935. The growth pattern and daily
rhythm for the incisor of the rat. Anatomical Record 63, 325–
333.
Schowalter, D. B., Harder, L. D., and Treichel, B. H. 1978. Age
composition of some vespertilionid bats as determined by dental
annuli. Canadian Journal of Zoology - Revue Canadienne de
Zoologie 56, 355–358. [cited in Klevezal 1996]
Schroeder, E. E. and Basket, T. S. 1968. Age estimation, growth
rates, and population structure in Missouri bullfrogs. Copeia
(3), 588–592.
Senning, W. C. 1940. A study of age determination and growth of
Necturus maculosus based on the parasphenoid bone. American
Journal of Anatomy 66, 483–494. [Smirina 1974 says this is the
first report on amphibian growth layers.]
Sheffield (no date) The Department of Archaeology and Prehistory
at University of Sheffield online glossary. http://www.shef.ac.uk/
uni/academic/A-C/ap/thin_sections/glossary.html. Retrieved 3
December 2002.
Singh, I. J. and Gunberg, D. L. 1971. Quantitative histology of
changes with age in rat bone cortex. Journal of Morphology
133, 241–252.
Skedros, J. G. 2001. Do osteon population densities reflect differ-
ences in fatigue history between limb bones of cursorial mam-
mals? Bone 28 (5 [Supp.]), S2000 – Abstract P 446 T.
Skyscan (no date) Skyscan. http://www.skyscan.be. Retrieved 16
December 2002.
Smirina, E. M. 1972. [Annual bands in the bones of Rana
temporaria]. Zoologicheskii Zhurnal 51, 1529–1534.
Smirina, E. M. 1974. [Prospects of age determination by bone layers
in Reptilia]. Zoologicheskii Zhurnal 53, 111–117.
Smirina, E. M., Klevezal, G. A. and Berger, L. 1986. [Experimental
study of annual layer formation in bones of amphibians].
Zoologicheskii Zhurnal 65, 1526–1534.
Smirina, E. M. and Makarov, A. N. 1987. [On ascertainment of
accordance between the number of layers in tubular bone of
amphibians and the age of individuals]. Zoologicheskii Zhurnal
66, 599–604.
Smirina, E. M., Serbinova, I. A. and Makarov, A. N. 1994. [Some
complicated cases of age determination using the annual layers
of bones in amphibians (at the example of long-tailed salamander
Onychodactylus fischeri)]. Zoologicheskii Zhurnal 73 (10), 72–
81.
Smitmans, L., Raum, K., Brandt, J. and Klemenz, A. 2000. Vari-
ations in the microstructural acousto-mechanical properties of
cortical bone revealed by a quantitative acoustic microscopy
study, in Schneider, S. C., Levy, M. and McAvoy, B. R. (eds.)
2000 IEEE Ultrasonics Symposium. Proceedings. An Intern-
ational Symposium / IEEE Ultrasonics, Ferroelectr., & Freq-
uency Control Soc., 22–25 Oct. 2000, in 2000 IEEE Ultrasonics
Symposium. Proceedings. An International Symposium.
Piscataway, NJ, US. IEEE 2, 1379–1382.
Smirnov, V. S. 1960. [Age determination and age relationship in
mammals with special reference to squirrel, muskrat, and five
species of carnivores]. Trudy Instituta Biologii (Akademija Nauk
SSSR, Ural’skij Filial) 14, 97–112. [cited in Klevezal 1996].
Stallibrass, S. 1982. The use of cement layers for absolute ageing of
mammalian teeth: A selective review of the literature, with
suggestions for further studies and alternative applications, pp.
109–126 in Wilson, B., Grigson, C. and Payne, S. (eds), Ageing
and Sexing Animal Bones from Archaeological Sites (BAR
British Series 109). Oxford: British Archaeological Reports.
Stanek, J. 1970. Rate of osteon growth in the diaphysis of long

39Using Osteohistology for Ageing and Sexing
bones of the young dog. Folia Morphologica 18, 381–388.
Stout, S. D., Brunsden, B. S., Hildebolt, C. F., Commean, P. K.,
Smith, K. E. and Tappen, N. C. 1999. Computer-assisted 3D
reconstruction of serial sections of cortical bone to determine the
3D structure of osteons. Calcified Tissue International 5, 280–
284.
Stutz, A. J. 2002. Polarizing microscopy identification of chemical
diagenesis in archaeological cementum. Journal of Archaeo-
logical Science 29, 1327–1347.
Sullivan, B. K. and Fernandez, P. J. 1999. Breeding activity,
estimated age-structure, and growth in Sonoran Desert anurans.
Herpetologica 55, 334–343.
Suzuki, H. K. 1963. Studies on the osseous system of the slider
turtle. Annals, New York Academy of Sciences 109, 351–410.
Szulc, P., Marchand, F., Duboeuf, F. and Delmas, P. D. 2000. Cross-
sectional assessment of age-related men: The MINOS study. Bone
26, 123–129.
Tappen, N. C. 2001. Studies of bone organization in three di-
mensions. American Journal of Physical Anthropology Annual
Meeting Issue, Supplement 32, 147. [abstract]
Teitelbaum, S. A. 2000. Bone resorption by osteoclasts. Science
289, 1504–1508.
Timlin, J. A., Garden, A., Morris, M. D., Rajachar, R. M. and Kohn,
D. H. 2000. Raman spectroscopic imaging markers for fatigue-
related microdamage in bovine bone. Analytical Chemistry 72,
2229–2236.
Tonna, E. A. 1966. A study of osteocyte formation and distribution
in aging mice complemented with H³-proline autoradiography.
Journal of Gerontology 21, 124–130.
Trivers, K. F. and Armelagos, G. J. 1997. Double zonal osteons and
HDI bodies in bone remodelling and modeling ancient Nubia.
American Journal of Physical Anthropology Supplement 24,
229–230. [abstract]
Trunova, Y. E. and Klevezal, G. A. 1999. [Interspecific variation in
the records of winter hibernation in incisor dentine of rodents].
Zoologicheskii Zhurnal 78, 1455–1464.
Trunova, Y. E., Lobkov, V. A. and Klevezal, G. A. 1999. The record
of the reproductive cycle in the incisor dentine of spotted souslik
Spermophilus Suslicus. Acta Theriologica 44, 161–171.
Vajda, E. G., Kneissel, M., Muggenburg, B. and Miller, S. C. 1999.
Increased intracortical bone remodelling during lactation in
beagle dogs. Biology of Reproduction 61, 1439–1444.
Van Neer, W. 1993. Limits of incremental growth in seasonality
studies: The example of the clariid pectoral spines from the
Byzantino-Islamic site of Apamea (Syria, 6th-7th century A.D.).
International Journal of Osteoarchaeology 3, 119–127.
Van Neer, W., Ervynck, A., Bolle, L., Millner, R. and Rijnsdorp, A.
2002a. Fish otoliths and their relevance to archaeology: An
analysis of Medieval, post-Medieval and recent material of plaice,
cod and haddock from the North Sea. Environmental
Archaeology 7, 65–81.
Van Neer, W., Noyen, K., De Cupere, B. and Beuls, I. 2002. On the
use of endosteal layers and medullary bone from domestic fowl
in archaeozoological studies. Journal of Archaeological Science
29, 123–134.
Van Soest, R. W. M. and Van Utrecht, W. L. 1971. The layered
structure of bones of birds as a possible indication of age.
Bijdragen tot de Dierkunde 41, 61–66. [cited in Van Neer et al.
1993 as the first attempt to use growth rings for ageing birds]
Victor, R. C. and Brothers, E. B. 1982. Age and growth of the
fallfish (Semotilus corporalis) with daily otolith increments as a
method of annulus verification. Canadian Journal of Zoology
60, 2543–2550.
Walker, W. F. and Liem, K. F. 2000. Functional anatomy of the
vertebrates: An evolutionary perspective. Forth Worth:
Saunders College Publications.
Warren, J. W. 1963. Growth zones in the skeleton of recent and
fossil vertebrates. Unpublished M.S. thesis, University of
California at Los Angeles. [cited in de Buffrénil and Buffetaut
1981]
Weaver, D. S., Jerome, C. P. and Peterson, P. 2001. Age-related
bone loss in nonhuman primates: Results and models. American
Journal of Physical Anthropology Supplement. 32, 161.
[abstract]
Williams, B. G. 2000. Aspects of bone quality in the broiler chicken.
Unpublished Ph.D. thesis, Glasgow University.
Wilson, J. W. 1994. Histological techniques, pp. 205–235 in Leiggi,
P. and May, P. (eds), Vertebrate Paleontological Techniques,
vol. 1. Cambridge: Cambridge University Press.
Winks, C. S. and Felts, W. J. L. 1980. Effects of castration on the
bone structure of male rats: A model of osteoporosis. Calcified
Tissue International 32, 77–82.
Yoshino, M., Imaizumi, K., Miyasaka, S. and Seta, S. 1994.
Histological estimation of age at death using microradiographs
of humeral compact bone. Forensic Science International 64,
191–198.
Kim Dammers
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