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Behavioral Biology of
Laboratory Animals
Behavioral Biology of
Laboratory ­Animals

edited by
Kristine Coleman and Steven J. Schapiro
First edition published 2022
by CRC Press
6000 Broken Sound Parkway NW, Suite 300, Boca Raton, FL 33487-2742

and by CRC Press

2 Park Square, Milton Park, Abingdon, Oxon, OX14 4RN

© 2022 Taylor & Francis Group, LLC

CRC Press is an imprint of Taylor & Francis Group, LLC

Reasonable efforts have been made to publish reliable data and information, but the author and publisher cannot assume responsibility for the
validity of all materials or the consequences of their use. The authors and publishers have attempted to trace the copyright holders of all material
reproduced in this publication and apologize to copyright holders if permission to publish in this form has not been obtained. If any copyright
material has not been acknowledged please write and let us know so we may rectify in any future reprint.

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Center, Inc. (CCC), 222 Rosewood Drive, Danvers, MA 01923, 978-750-8400. For works that are not available on CCC please contact
­mpkbookspermissions@tandf.co.uk

Trademark notice: Product or corporate names may be trademarks or registered trademarks and are used only for identification and explanation
without intent to infringe.

ISBN: 978-0-367-02923-4 (hbk)

ISBN: 978-1-032-03435-5 (pbk)
ISBN: 978-0-429-01951-7 (ebk)

DOI: 10.1201/9780429019517

Typeset in Times
by codeMantra
Contents

Preface ...........................................................................................................................................................................................vii
Editors ............................................................................................................................................................................................. ix
Contributors .................................................................................................................................................................................... xi

Part 1
1. Introduction to the Behavioral Biology of Laboratory Animals ........................................................................................ 3
Steven J. Schapiro and Kristine Coleman

2. Animal Behavior: An Introduction ...................................................................................................................................... 7

Kristine Coleman and Melinda A. Novak

3. Abnormal Behavior of Animals in Research Settings ...................................................................................................... 27

Melinda A. Novak and Jerrold S. Meyer

4. Utilizing Behavior to Assess Welfare ................................................................................................................................. 51

Daniel Gottlieb and Ori Pomerantz

5. An Overview of Behavioral Management for Laboratory Animals ............................................................................... 65

Steven J. Schapiro

Part 2
6. Behavioral Biology of Mice ................................................................................................................................................. 89
Aileen MacLellan, Aimée Adcock, and Georgia Mason

7. Behavioral Biology of Rats .................................................................................................................................................113

Sylvie Cloutier

8. Behavioral Biology of Guinea Pigs ....................................................................................................................................131

Gale A. Kleven

9. Behavioral Biology of Deer and White-Footed Mice, Mongolian Gerbils, and Prairie and Meadow Voles..............147
Kathleen R. Pritchett-Corning and Christina Winnicker

10. Behavioral Biology of Hamsters ....................................................................................................................................... 165

Christina Winnicker and Kathleen R. Pritchett-Corning

11. Behavioral Biology of Rabbits............................................................................................................................................173

Lena Lidfors and Kristina Dahlborn

12. Behavioral Biology of Ferrets.............................................................................................................................................191

Claudia M. Vinke, Nico J. Schoemaker, and Yvonne R. A. van Zeeland

13. Behavioral Biology of Dogs ............................................................................................................................................... 205

Laura Scullion Hall and Mark J. Prescott

14. Behavioral Biology of the Domestic Cat........................................................................................................................... 223

Judith Stella

v
vi Contents

15. Behavioral Biology of Pigs and Minipigs ......................................................................................................................... 243

Sandra Edwards and Nanna Grand

16. Behavioral Biology of Sheep .............................................................................................................................................. 261

Cathy M. Dwyer

17. Behavioral Biology of Cattle ............................................................................................................................................. 273

Clive Phillips

18. Behavioral Biology of Horses ............................................................................................................................................ 285

Janne Winther Christensen

19. Behavioral Biology of Chickens and Quail ...................................................................................................................... 299

Laura Dixon and Sarah Lambton

20. Behavioral Biology of the Zebra Finch .............................................................................................................................315

Samantha R. Friedrich and Claudio V. Mello

21. Behavioral Biology of Zebrafish ........................................................................................................................................331

Christine Powell, Isabel Fife-Cook, and Becca Franks

22. Behavioral Biology of Amphibians ................................................................................................................................... 345

Charlotte A. Hosie and Tessa E. Smith

23. Behavioral Biology of Reptiles .......................................................................................................................................... 361

Dale F. DeNardo

24. Behavioral Biology of Marmosets..................................................................................................................................... 377

Arianna Manciocco, Sarah J. Neal Webb, and Michele M. Mulholland

25. Behavioral Biology of Squirrel Monkeys ......................................................................................................................... 395

Anita I. Stone and Lawrence Williams

26. Behavioral Biology of Owl Monkeys ................................................................................................................................ 409

Alba García de la Chica, Eduardo Fernandez-Duque, and Lawrence Williams

27. Behavioral Biology of Capuchin Monkeys ....................................................................................................................... 421

Marcela Eugenia Benítez, Sarah F. Brosnan, and Dorothy Munkenbeck Fragaszy

28. Behavioral Biology of Macaques....................................................................................................................................... 437

Paul E. Honess

29. Behavioral Biology of Vervets/African Green Monkeys................................................................................................. 475

Matthew J. Jorgensen

30. Behavioral Biology of Baboons ......................................................................................................................................... 495

Corrine K. Lutz

Part 3 Selected Ethograms .................................................................................................................................................513

Index ............................................................................................................................................................................................ 529

Preface
The Behavioral Biology of Laboratory Animals is designed
to provide readers with knowledge about the behavioral biol-
ogy of animals that are common subjects in research set-
tings. Knowing the natural behavior of animals is a critical
part of modern animal care practices. Each species evolved
under specific environmental conditions resulting in unique
behavioral patterns, many of which are maintained in captiv-
ity even after generations of breeding. Thus, factors such as
how individuals obtain food and avoid predation, whether they
maintain territories, how they interact and communicate with
conspecifics, and how they care for their young are important
considerations when providing appropriate housing options for
laboratory animals. Knowing the behavioral biology allows
facilities to provide conditions that functionally simulate their
environment and encourage the expression of natural behavior.
In addition, knowing behaviors that are normal for the spe-
cies can help those caring for them recognize behaviors of
concern. Certain vocalizations or behavioral patterns, such
as scratching in nonhuman primates and dogs, can indicate
that the animal is feeling anxious. Abnormal behaviors such
as excessive stereotypy can indicate reduced welfare. For this
reason, the Guide for the Care and Use of Laboratory Animals
suggests that “Personnel responsible for animal care and hus-
bandry should receive training in the behavioral biology of the
species they work with to appropriately monitor the effects of
enrichment as well as identify the development of adverse or
abnormal behaviors” (National Resource Council 2011, p. 53).
This book provides behavioral information for many,
although not all, common laboratory animals. Because not
every reader has a background in animal behavior, the book is
broken into three parts to cover both broad aspects of animal
behavior and how to maintain animals in captivity, as well as
the behavioral biology of specific laboratory animals.
Part 1 of this book covers some of the basics of animal
behavior and how behavior can be used to assess welfare.
The heart of the book is the next part, which focuses on
the behavioral biology of specific laboratory animal spe-
cies, or group of species. Each chapter discusses the natural
behavior of the animal, and because the behaviors animals
express in captivity can be somewhat different than those
found in their wild counterparts, each includes a section on
common captive behavior, including those considered both
normal and abnormal. Additionally, the chapters in this
part of the book provide information on ways to maintain
the behavioral health of the animals. The third part of the
book contains ethograms (lists of behaviors and their defi-
nitions) for many of the species. These ethograms will pro-
vide a quick reference for care staff, researchers, and casual
observers alike.
As you look through this book, you will read about the fasci-
nating and unique behaviors displayed by each of the animals.
Working with these animals is a great privilege, accompanied,
of course, by a great responsibility. We are so grateful to the
authors of this book for sharing their knowledge of, and love
for, the animals with which they work. We hope that not only
does this book help you to learn more about the species in your
care, but also to better honor the contributions made by the
animals to research endeavors.
vii
Editors
Kristine Coleman, PhD, is an associate professor in the
Division of Comparative Medicine, and Head of the Behavioral
Services Unit at the Oregon National Primate Research Center
(ONPRC), Oregon Health & Science University. Dr. Coleman
received her PhD in behavioral ecology from Binghamton
University, where she studied individual differences in tem-
perament in pumpkinseed sunfish. She went to the Oregon
Regional (now National) Primate Research Center for her
postdoctoral training and never left. Since 2001, she has over-
seen the ONPRC behavioral management program, where she
studies ways to improve the psychological well-being of labo-
ratory macaques.
Dr. Coleman has coauthored over 150 papers, book chapters,
and abstracts on various topics in the field of animal behavior
and management. She regularly teaches courses and work-
shops on behavioral management topics. For the past 5 years,
she has taught a course on the behavioral biology of labora-
tory animals in the Eastern Virginia Medical School’s Master
of Laboratory Animal Science program. Dr. Coleman is a
member of a number of animal behavior societies and is cur-
rently the treasurer and co-chair of the Member and Finance
Committee of the American Society of Primatologists. In
addition, Dr. Coleman is vice-chair of the ONPRC IACUC,
and an ad hoc specialist with AAALAC International.
Steven J. Schapiro, PhD, is an associate professor of compar-
ative medicine in the Department of Comparative Medicine at
the Michale E. Keeling Center for Comparative Medicine and
Research of The University of Texas MD Anderson Cancer
Center. Dr. Schapiro earned his PhD from the University of
California at Davis in 1985 after receiving his BA in behav-
ioral biology from Johns Hopkins University. He completed
a postdoctoral research fellowship at the Caribbean Primate
Research Center of the University of Puerto Rico.
In 1989, he joined the Department of Comparative Medicine
at MD Anderson’s Keeling Center and has been there ever
since. In 2009, Dr. Schapiro was designated an honorary pro-
fessor in the Department of Experimental Medicine at the
University of Copenhagen, Denmark. He is a founding fac-
ulty member of both the Primate Training and Enrichment
Workshop and the Primate Behavioral Management
Conference, educational programs conducted at the Keeling
Center that have reached over 1,000 individuals from primate
facilities around the globe. He is the founder and organizer
of Project Monkey Island, a rescue and restoration mission
for the monkeys and people associated with Cayo Santiago in
Puerto Rico.
Dr. Schapiro has coauthored almost 200 peer-reviewed
papers and book chapters examining various aspects of non-
human primate behavior, management, and research. He has
also edited the three volumes of the third edition, and the fourth
edition of the Handbook of Laboratory Animal Science along
with Jann Hau, and served as the sole editor of the Handbook
of Primate Behavioral Management. He has also coedited one
issue of the ILAR Journal.
Dr. Schapiro has participated in international meetings and
courses on primatology, behavioral management, and labo-
ratory animal science in North America, Europe, Asia, and
Africa. He is a member of a number of primatology and ani-
mal behavior societies and served as the treasurer and vice
president for membership of the International Primatological
Society for many years. He is also a past president, former
treasurer, and former meeting coordinator of the American
Society of Primatologists, as well as an honorary member
of the Association of Primate Veterinarians. Dr. Schapiro is
an advisor or consultant for a number of primate facilities
that produce, manage, and conduct research with nonhuman
primates in the United States and abroad.
ix
Contributors

Aimée Adcock Cathy M. Dwyer

Formerly Department of Animal Bioscience Animal Behaviour and Welfare
University of Guelph Scotland’s Rural College
Guelph, Ontario, Canada Penicuik, Midlothian, United Kingdom

Sandra Edwards
Marcela Eugenia Benítez
School of Natural and Environmental Sciences
Department of Anthropology
Newcastle University
Emory University
Newcastle, United Kingdom
Atlanta, Georgia, United States
Eduardo Fernandez-Duque
Sarah F. Brosnan Department of Anthropology
Departments of Psychology and Philosophy Yale University
Language Research Center New Haven, Connecticut, United States
Center for Behavioral Neuroscience and
Neuroscience Institute Isabel Fife-Cook
Georgia State University Department of Environmental Studies
Atlanta, Georgia, United States New York University
New York, New York, United States
Janne Winther Christensen
Department of Animal Science Dorothy Munkenbeck Fragaszy
Aarhus University Department of Psychology
Aarhus, Denmark University of Georgia
Athens, Georgia, United States
Sylvie Cloutier
Animal Behavior Scientist Becca Franks
Ottawa, Ontario, Canada Department of Environmental Studies
New York University
Kristine Coleman New York, New York, United States
Oregon National Primate Research Center
Oregon Health & Science University Samantha R. Friedrich
Beaverton, Oregon, United States Department of Behavioral Neuroscience
Oregon Health & Science University
Kristina Dahlborn Portland, Oregon, United States
Department of Anatomy Physiology and Biochemistry
Swedish University of Agricultural Sciences Daniel Gottlieb
Uppsala, Sweden Oregon National Primate Research Center
Oregon Heath & Science University
Alba García de la Chica Beaverton, Oregon, United States
Universidad de Barcelona
Barcelona, Spain Nanna Grand
Scantox A/S
Dale F. DeNardo Lille Skensved, Denmark
Department of Animal Care and Technologies
School of Life Sciences Laura Scullion Hall
Arizona State University Refining Dog Care and
Tempe, Arizona, United States University of Stirling
Stirling, United Kingdom
Laura Dixon
Animal Behaviour and Welfare
Scotland’s Rural College
Penicuik, Midlothian, United Kingdom

xi
xii Contributors

Paul E. Honess Jerrold S. Meyer

School of Veterinary Medicine and Science Department of Psychological and Brain Sciences
University of Nottingham University of Massachusetts-Amherst
Nottingham, United Kingdom Amherst, Massachusetts, United States

Charlotte A. Hosie Michele M. Mulholland

Department of Biological Sciences Department of Comparative Medicine
University of Chester The University of Texas MD Anderson Cancer Center
Chester, United Kingdom Bastrop, Texas, United States

Sarah J. Neal Webb

Matthew J. Jorgensen Department of Comparative Medicine
Department of Pathology The University of Texas MD Anderson Cancer Center
Section on Comparative Medicine Bastrop, Texas, United States
Wake Forest School of Medicine
Winston-Salem, North Carolina, United States Melinda A. Novak
Department of Psychological and Brain Sciences
Gale A. Kleven University of Massachusetts, Amherst
Department of Psychology Amherst, Massachusetts, United States
Behavioral Neuroscience Area
Wright State University Clive Phillips
Dayton, Ohio, United States Curtin University Sustainability Policy Institute
Curtin University
Sarah Lambton Perth, Western Australia, Australia
Bristol Veterinary School
University of Bristol Ori Pomerantz
Langford, North Somerset, United Kingdom California National Primate Research Center
University of California, Davis
Lena Lidfors Davis, California, United States
Department of Animal Environment and Health
Swedish University of Agricultural Sciences Christine Powell
Skara, Sweden Department of Environmental Studies
New York University
Corrine K. Lutz New York, New York, United States
Animal Behavior Scientist
San Antonio, Texas, United States Mark J. Prescott
National Centre for the Replacement, Refinement and
Aileen MacLellan Reduction of Animals in Research (NC3Rs)
Department of Integrative Biology London, United Kingdom
University of Guelph
Guelph, Ontario, Canada Kathleen R. Pritchett-Corning
Office of Animal Resources
Arianna Manciocco Harvard University Faculty of Arts and Sciences
Istituto di Scienze e Tecnologie della Cognizione Department of Comparative Medicine
Consiglio Nazionale delle Ricerche Cambridge, Massachusetts, United States
Rome, Italy and
University of Washington
Georgia Mason Seattle, Washington, United States
Department of Integrative Biology
University of Guelph Steven J. Schapiro
Guelph, Ontario, Canada Department of Comparative Medicine
The University of Texas MD Anderson Cancer Center
Claudio V. Mello Bastrop, Texas, United States
Department of Behavioral Neuroscience and
Oregon Health & Science University Department of Experimental Medicine
Portland, Oregon, United States University of Copenhagen
Copenhagen, Denmark
Contributors
Nico J. Schoemaker
Division of Zoological Medicine
Department of Clinical Sciences
Faculty of Veterinary Medicine
University of Utrecht
Utrecht, The Netherlands
Tessa E. Smith
Department of Biological Sciences
University of Chester
Chester, United Kingdom
Judith Stella
Good Dog, Inc.
New York, New York, United States
Anita I. Stone
Department of Biology
California Lutheran University
Thousand Oaks, California, United States
xiii
Yvonne R.A. van Zeeland
Division of Zoological Medicine
Department of Clinical Sciences
Faculty of Veterinary Medicine
University of Utrecht
Utrecht, The Netherlands
Claudia M. Vinke
Division of Animals in Science and Society
Department of Population Health Sciences
Faculty of Veterinary Medicine
University of Utrecht
Utrecht, The Netherlands
Lawrence Williams
Department of Comparative Medicine
The University of Texas MD Anderson Cancer Center
Bastrop, Texas, United States
Christina Winnicker
Research Office of Animal Welfare Ethics Strategy & Risk
GlaxoSmithKline
Collegeville, Pennsylvania, United States
Part 1
1
Introduction to the Behavioral Biology of Laboratory Animals
Steven J. Schapiro
The University of Texas MD Anderson Cancer Center
University of Copenhagen
Kristine Coleman
Oregon National Primate Research Center
CONTENT
References ........................................................................................................................................................................................ 5
Welcome to the Behavioral Biology of Laboratory Animals
(BBLA). This volume contains 30 chapters divided into three
sections, which all focus on aspects of the behavioral biology
of animals that are often/frequently/sometimes/occasionally
found in laboratories and related research settings. The overall
goal of the BBLA is to provide you, someone working in a
maintenance, care, and/or research program that involves lab-
oratory animals, with information about the way the animals
live in the wild, and the way that they should live in captive
research settings. We hope that the contents of the chapters
(descriptions, data, guidance, resources, and recommenda-
tions) will help you understand your animals better, allow you
to refine the care and treatment that your animals receive, and
improve the well-being, welfare, and wellness of your animals.
If the data show that when intact adult male New Zealand
White (NZW) rabbits are housed together in captivity (a social
situation that is extremely unlikely to be observed in nature),
they are quite likely to fight, then, unfortunately, captive, intact
adult male NZWs should probably be housed individually
(Lidfors and Dahlborn, 2021). At the opposite end of the spec-
trum, if the data show that zebra finches that are housed singly
in captivity (again, a social situation that is extremely unlikely
to be observed in the wild) fail to prosper, then (fortunately)
captive zebra finches should be housed socially (in pairs or
flocks; Friedrich and Mello, 2021).
The BBLA includes an entire section (Part 2; 25 chapters) on
the behavioral biology of specific taxonomic groups (e.g., mice,
zebrafish, zebra finches, reptiles, macaques), preceded by five
introductory chapters that focus on the science of studying
behavior as a discipline and its impact on our understanding
of the species with which we work. The book concludes with a
section that includes ethograms for some/many of the species
discussed. The ethograms in Part 3 should provide a relatively
centralized resource for those interested in understanding, and
potentially quantifying, baboon, bird, or bunny behavior.
Most of the chapter authors in this volume study and/or
work with animals in “research” settings; therefore, the infor-
mation contained in their chapters is likely to be most relevant
to animals living in such settings. However, many of the guide-
lines and recommendations contained in this book will also
be valuable to those managing and working with animals in
other environments, including zoological parks, aquaria, and
sanctuaries.
The BBLA evolved as we each attempted to teach courses
on the behavior of laboratory animals (SJS at the University
of Copenhagen and KC at Eastern Virginia Medical School).
Neither of us could find a single resource that addressed all of
the issues that we felt were important to impress upon our stu-
dents. KC, being a person of action, decided that we could fill
this resource void by putting together a volume that addressed
the issues that we thought would be most important to those
working with laboratory animals. SJS came along for the ride,
as he has certain organizational skills and connections that
helped this project come to fruition. So, here you are, reading
the result of a slightly delayed (partially due to COVID-19),
but hopefully, successful, collaboration among the publish-
ers, editors, and the authors. We are both reasonably seasoned
observers of animal behavior, but we each learned quite a bit
from working on these chapters. Some of our favorites include
popcorning and rumblestrutting in guinea pigs (Kleven,
2021), the weasel war dance in ferrets (Vinke et al., 2021),
and clumping in zebra finches (Friedrich and Mello, 2021).
We hope that you will learn a few things and discover a few
favorites as well.
Part 1 of the BBLA includes five chapters by Schapiro and
Coleman; Coleman and Novak; Novak and Meyer; Gottlieb
and Pomerantz; and Schapiro; respectively. Schapiro’s and
Coleman’s chapter, which you are currently reading, estab-
lishes the motivation for the book and describes the chapters
herein. The chapter by Coleman and Novak explains the role
that animal behavior research plays in the selection and utiliza-
tion of animals in laboratory research programs. Their chapter
establishes the fundamental role that studying and understand-
ing animal behavior plays in scientific research that involves
animals. Pay particular attention to the discussion of the dif-
ferences between animal and human sensation/perception.
3
4 Behavioral Biology of Laboratory Animals
This chapter has much to offer those responsible for study-
ing and caring for laboratory animals. In Chapter 3, Novak
and Meyer provide a brief review of abnormal behaviors in
laboratory animals, with an emphasis on nonhuman primates
(NHPs). While colony managers, veterinarians, researchers,
and behaviorists make every attempt to establish captive envi-
ronments that discourage the performance of abnormal activi-
ties, we are still learning how to prevent abnormal behavior. It
is important that those of us working with animals in the labo-
ratory have some understanding of behavioral abnormalities,
the conditions that may yield them, and potential treatments
to address them. Gottlieb and Pomerantz, in Chapter 4, then
provide several behaviorally oriented perspectives that can be
used to assess welfare in laboratory animals. They describe
three approaches that should be helpful in determining
whether strategies employed to maintain and enhance the wel-
fare of laboratory animals are successful. Part 1 closes with a
discussion of behavioral management by Schapiro, again with
an emphasis on NHPs. Strategies to stimulate species-typical
behavior and to minimize abnormal behavior are outlined in
general terms, as each of the taxon-specific chapters in Part
2 contains several sections intended to address the ways that
questions related to behavioral management are being handled
in the animals included in the chapter. All five of these chap-
ters help to establish a foundation for the taxon-specific infor-
mation contained in the chapters that follow.
Part 2 of the BBLA comprises 25 chapters describing the
behavioral biology of different taxonomic groups of animals.
It should be obvious that we could not include every animal
species that is involved in laboratory research. It should also
be obvious that not every author we asked to write a chapter
was able to complete the task. We are missing a few chapters
that we would have liked to include (e.g., cephalopods, addi-
tional fish and avian species), but overall, we feel as though we
included many of the most important animal species involved
in research.
Some chapters cover a relatively small amount of taxonomic
real estate (e.g., Chapter 8 by Kleven on guinea pigs), while
others cover a considerable range of genera (e.g., Chapter 22
by Hosie and Smith on amphibians, Chapter 23 by DeNardo on
reptiles) and/or species (Chapter 28 by Honess on macaques).
We asked the authors of the chapters in this section to adhere
to a particular format, beginning with a discussion of the typi-
cal research involvement of the taxa, followed by the natural
history (e.g., ecology, social organization, feeding behavior,
communication) of the animals and their common behaviors
in captivity (normal and abnormal). Authors then present sec-
tions on ways to maintain behavioral health and stimulate
species-typical behavior in captive settings. After brief discus-
sions of situations that are special to that taxon, the chapters
conclude with recommendations, resources, and, of course,
references. The authors of these chapters have quite a bit of rel-
evant experience, and reading their work should provide you
with a wealth of useful information on the behavioral biology
of these taxonomic groups.
Part 2 starts with a set of six chapters on rodents and lago-
morphs, and begins with a fantastic chapter (Chapter 6) by
MacLellan and colleagues on the behavioral biology of mice.
As mice comprise the taxonomic group that is most frequently
involved in research laboratories, it makes sense to start with
them. There is much to be learned in this chapter. Chapter 7,
by Cloutier, addresses the behavioral biology of rats, another
popular and well-utilized taxon. A great deal is known about
rat behavior in captive settings, but relatively little is known
about the way they function in natural habitats. Chapter 8 by
Kleven on guinea pigs comes next, with “popcorning” and
the “rumblestrut”, new behaviors for SJS and KC to add to
our internal ethograms of animal behaviors. Guinea pigs are
also known to “stampede”, which creates a fun visual image.
Pritchett-Corning and Winnicker describe gerbils, voles, and
deer mice in Chapter 9, including arboreal Peromyscus and
monogamous Microtus. Winnicker and Pritchett-Corning
change author order for Chapter 10, in which they discuss
hamsters. As those who have worked with hamsters know,
they can be a bit grumpy, when rudely awakened. This is
the final rodent chapter in this part. Lidfors and Dahlborn
(Chapter 11) then address the intricacies of socially housing
intact, adult male rabbits in the context of their natural history,
among other topics.
Three chapters focusing on the behavioral biology of carni-
vores (ferrets by Vinke and colleagues, Chapter 12; Hall and
Prescott on dogs, Chapter 13; and Stella on cats, Chapter 14)
follow the rodent/lagomorph chapters, and one must pay par-
ticular attention to the weasel war dance performed by ferrets.
Among “humans’ best friends”, which have olfactory systems
that are a million to a billion times more sensitive than those of
humans, selectively and purpose-bred beagles are especially
important. It is also interesting to note that the cat is one of
only a few types of animals in which the wild ancestor/relative
is solitary, and the “domesticated” members of the species are
quite social.
A series of five chapters on farm-type animals comes next,
beginning with pigs and minipigs by Edwards and Grand
(Chapter 15). The behavior of domestic pigs and mini-pigs dif-
fers little from that of their wild relatives, and considerable
attention must be paid to weaning-related transitions and the
provision of nest-building opportunities for pigs in research.
Also, it should be noted that mother pigs do not lick their
infants, a bit of an anomaly among mammals. Dwyer discusses
the behavioral biology of sheep in Chapter 16, explaining that
they are highly social, engage in many types of allomimetic
behavior, and are, surprisingly, highly trainable. The behav-
ioral biology of cattle that participate in research is described
by Phillips (Chapter 17), followed by horses in Chapter 18
(Christensen). Horses are interesting, in that (1) negative rein-
forcement training (rather than positive reinforcement) is typi-
cally employed in practical situations, and (2) the mare-foal
bond is extremely important for normal development. This
subsection concludes with Chapter 19 on the behavioral biol-
ogy of chickens by Dixon and Lambton, the first of only two
chapters on avian taxa. The second avian contribution is a
wonderful chapter (Chapter 20) by Friedrich and Mello that
beautifully describes the behavioral biology of zebra finches
(affectionately referred to by some as “zebbies”), including
their vocalizations and their tendency to clump.
The next three chapters continue the departure from mam-
malian species, with chapters (21 by Powell and colleagues) on
zebrafish, (22 by Hosie and Smith) on amphibians, and (23 by
Introduction
DeNardo) on reptiles. Not only is the zebrafish chapter highly
informative, all of the impressive illustrations were drawn by
one of the authors. While Powell and colleagues were asked to
focus their efforts on only the increasingly research-relevant
zebrafish, Hosie and Smith and DeNardo were given the
monumental tasks of addressing the behavioral biology of the
multileveled taxonomic groups of amphibians and reptiles,
respectively. They have each done a fantastic job of present-
ing important information on the behavioral biology of frogs,
toads, and salamanders, and of snakes, lizards, crocodilians,
and turtles/tortoises, respectively.
Part 2 concludes with seven chapters on various “species”
of NHPs; four New World monkeys and three Old World mon-
keys. Chapter 24, by Manciocco and colleagues, discusses
marmosets, with an emphasis on the common marmoset, an
increasingly important NHP species in laboratory research,
given its amenability to CRISPR-Cas9 technology. Stone and
Williams, in Chapter 25, report on squirrel monkeys, a taxon
with a long history of studies in the wild and the laboratory.
In Chapter 26, Garcia de la Chica and colleagues discuss owl
(night) monkeys, primates that are primarily nocturnal (as
their common name implies), making them quite difficult to
observe in the wild. Studies of owl monkeys in captive settings
(making use of altered light cycles) have truly informed our
understanding of their natural behavioral biology. This chap-
ter is the only one in the book to discuss nocturnal NHPs, but
not the only chapter to discuss nocturnal animals (e.g., mice,
rats). Benitez and colleagues discuss the so-called “capuchin”
monkeys (their taxonomy is somewhat in flux) in Chapter 27.
These extremely interesting and intelligent animals have made
important contributions toward answering a variety of psy-
chological and anthropological research questions. Chapter
28 (by Honess) is another chapter devoted to the behavioral
biology of a very large taxonomic group, the macaques. This
chapter does an excellent job of providing information on the
behavioral biology of a wide range of macaque species, while
emphasizing the two most frequently encountered NHPs in
laboratory research (Macaca mulatta and M. fascicularis).
The penultimate chapter (Chapter 29 by Jorgensen) focuses
on another group that is in a bit of taxonomic flux, “vervet
monkeys”. Vervets are involved less frequently in laboratory
research than macaques, and Jorgensen clearly explains that
the behavior in nature of African vervets differs considerably
from the natural behavior of the primarily Asian macaques.
Therefore, vervet monkeys cannot be managed in the same
way as macaques. Part 2 ends with a chapter about baboons
(Chapter 30 by Lutz), another NHP species with a long research
history in the wild and the laboratory, and a recent history that
involves taxonomic questions. This excellent chapter is a fit-
ting way to end Part 2. In an odd twist of fate, Lutz’s chapter
was the first one received by us, but is the last one out to you.
Part 3 of the BBLA is simply a collection of ethograms for
a selection of the taxa included in Part 2. We felt that it would
be extremely useful for you to have a centralized resource
that contains verified ethograms for many of the species that
most frequently live in laboratory settings and participate in
research projects. Over the years, we have searched for this
type of resource, without much luck. Hopefully, you will find
Part 3 to be a useful addition to this volume.
5
There are just three more things to quickly mention before
you venture off into the biological landscape of the BBLA.
1. You will encounter some common themes, tech-
niques, and terminology as you read the chapters in
this volume. These include in no particular order:
temperament/personality, behavioral assessments,
functional simulations, functionally appropriate
captive environments, prevention vs. cure, refine-
ments, research–management synergisms, positive
reinforcement training, enhancing the definition of
animal models, minimizing confounds, and better
scientific data. Please pay attention to these points
when you encounter them.
2. In a similar vein, you will read about similar things
that are, at times, labeled in slightly different ways
in different parts of the book. We tried to establish a
consistent framework across chapters, but sometimes
it did not make sense to change what the authors had
written. Keep your eyes open for similarities and dif-
ferences in terms used. For instance, the definitions
for abnormal behaviors or types of environmental
enrichment may differ slightly across chapters, but
there are really many more similarities than differ-
ences in these cases.
3. And finally, do not forget that the goal of
Behavioral Biology of Laboratory Animals is to
provide you with research-based information, from
experts, that you can use to better understand, and
to benefit, the animals that you care for, and work
with, in your “laboratories”. Understanding their
behavioral biology is good for the animals and
good for science, contributing to the optimization
of the welfare of the animals, and the reliability
and validity of the data. Behavioral biology as a
science is constantly evolving, as new methods,
techniques, and care strategies are designed and
implemented. We can always learn more about
our animals and do more to enhance their lives in
research settings.
REFERENCES
Friedrich SR, Mello CV. Behavioral biology of zebra finches
(Taeniopygia guttata). In Coleman K, Schapiro SJ, eds.
Behavioral Biology of Laboratory Animals. Boca Raton,
FL: CRC Press, Taylor & Francis Group; 2021:315–329.
Kleven GA. Behavioral biology of guinea pigs. In Coleman
K, Schapiro SJ, eds. Behavioral Biology of Laboratory
Animals. Boca Raton, FL: CRC Press, Taylor & Francis
Group; 2021:131–146.
Lidfors L, Dahlborn K. Behavioral biology of rabbits. In Coleman
K, Schapiro SJ, eds. Behavioral Biology of Laboratory
Animals. Boca Raton, FL: CRC Press, Taylor & Francis
Group; 2021:173–190.
Vinke CM, Schoemaker NJ, van Zeeland YRA. Behavioral biol-
ogy of ferrets. In Coleman K, Schapiro SJ, eds. Behavioral
Biology of Laboratory Animals. Boca Raton, FL: CRC
Press, Taylor & Francis Group; 2021:191–204.
2
Animal Behavior: An Introduction

Kristine Coleman
Oregon National Primate Research Center

Melinda A. Novak
University of Massachusetts, Amherst

CONTENTS
Introduction�������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������� 7
The Definition and Challenge of Animal Behavior������������������������������������������������������������������������������������������������������������������������� 8
Description Phase����������������������������������������������������������������������������������������������������������������������������������������������������������������������� 8
Quantification Phase������������������������������������������������������������������������������������������������������������������������������������������������������������������� 9
The Explanation Phase�������������������������������������������������������������������������������������������������������������������������������������������������������������� 10
Biologically Relevant Questions about Behavior�������������������������������������������������������������������������������������������������������������������������� 10
Proximate Questions (Mechanism and Development)�������������������������������������������������������������������������������������������������������������� 11
Ultimate Questions (Function and Evolutionary History)�������������������������������������������������������������������������������������������������������� 11
Bird Song as an Example of a Levels of Analysis Perspective������������������������������������������������������������������������������������������������� 11
Using the Scientific Method to Study Behavior���������������������������������������������������������������������������������������������������������������������������� 12
Concepts from Animal Behavior That Impact Laboratory Animal Care and Welfare������������������������������������������������������������������� 13
Ecology������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������� 13
Spatial Distributions������������������������������������������������������������������������������������������������������������������������������������������������������������ 13
Predation and the Landscape of Fear����������������������������������������������������������������������������������������������������������������������������������� 13
Social Interactions and Organization���������������������������������������������������������������������������������������������������������������������������������������� 14
Social Organization�������������������������������������������������������������������������������������������������������������������������������������������������������������� 14
Dominance Hierarchies�������������������������������������������������������������������������������������������������������������������������������������������������������� 15
Sexual Selection and Mating����������������������������������������������������������������������������������������������������������������������������������������������� 15
Foraging and Feeding Behavior������������������������������������������������������������������������������������������������������������������������������������������������ 17
Communication������������������������������������������������������������������������������������������������������������������������������������������������������������������������� 17
Learning������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������ 18
Individual Variation������������������������������������������������������������������������������������������������������������������������������������������������������������������� 20
Cautions: Our Limitations as Observers���������������������������������������������������������������������������������������������������������������������������������������� 21
Sensory Motor Capabilities������������������������������������������������������������������������������������������������������������������������������������������������������ 21
Anthropomorphism������������������������������������������������������������������������������������������������������������������������������������������������������������������� 21
Summary���������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������� 21
Acknowledgments�������������������������������������������������������������������������������������������������������������������������������������������������������������������������� 22
References�������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������� 22

rabbits freeze and/or flee to escape predation. Animals evolved

Introduction
If you were fortunate enough to have had time to take a walk in
your neighborhood today, you may have heard robins sing. Or,
perhaps you witnessed a squirrel run across a field to retrieve a
previously buried nut. You may have caught a rabbit off guard
and watched as he remained motionless before running away.
All of these behaviors, as well as others we may not notice, help
the animals survive in their world. Many bird species sing to
attract mates, squirrels hide (cache) nuts to consume later, and
to perform specific behaviors, such as the ones listed, that help
them survive and reproduce. As such, animals have a strong
drive to engage in these behaviors, even when they are living
in other environments, such as research facilities. Even geneti-
cally altered animals maintain behavioral instincts of their wild
counterparts. In order to properly care for these animals, it is
important that we understand their behavioral biology, a point
that is emphasized in regulatory and accreditatory documents,
including the eighth edition of the Guide for the Care and Use of
Laboratory Animals (National Resource Council 2011) and the

7
8 Behavioral Biology of Laboratory Animals
European Convention for the Protection of Vertebrate Animals
Used for Experimental and Other Scientific Purposes, Council
of Europe (ETS 123, Appendix A).
There are several reasons why understanding the behavioral
biology of animals is important to their care. As mentioned
above, animals evolved specific behavioral repertoires that
help them survive in their natural environment. When animals
are not able to perform these behaviors, it can cause stress and
anxiety, as well as the development of undesired behavior,
which can negatively impact their welfare (see Gottlieb and
Pomerantz 2021; Novak and Meyer 2021). Knowing the behav-
ioral biology of the animals is key to being able to provide
them the features of their environment that can best promote
these behaviors. For example, species that build nests, such
as mice, are generally provided with appropriate nest making
material. In addition, an individual’s behavior provides infor-
mation about its welfare state. As you will read in this book
(e.g., Gottlieb and Pomerantz 2021), the absence of normal
behavior, as well as the presence of abnormal behavior, can
indicate pain and/or distress in animals. Thus, understand-
ing normative behavior is key to assessing well-being. Lastly,
while we are perhaps biased, having chosen to devote our
careers to the study of animal behavior, we believe it is one of
the most fascinating scientific fields. As you read through the
chapters in this book, you will see that animals engage in some
pretty amazing behaviors.
This book examines the behavioral biology of animals that
participate in research. Each chapter in the main part of the book,
Part 2, examines how a specific animal – or group of animals –
behaves in its natural environment. What you will find, if you
read all of those chapters, is that while specifics vary, there are
a number of similarities as well. In order to survive, animals,
regardless of their size or speed, have to be able to find and pro-
cess food, while simultaneously avoiding being a food source for
someone else. Most of them try to find mates and many care
for offspring. These aspects of their behavior are often intercon-
nected and are related to the environment in which they live.
Equally important to understanding environmental factors
is understanding how animals perceive their environment.
An animal’s perceptual world, or umwelt, is closely tied to its
senses (von Uexküll 2010; Burnett 2011). Humans have five
senses, but we perceive our world largely through hearing
and vision. We hear sounds in a frequency range from about
20 Hz to 20 kHz, and see three colors (blue, green, and red).
However, this is not the same way other animals perceive their
world. Many animals hear sounds in ultrasonic ranges that we
cannot. Birds are tetrachromatic and see UV in addition to the
colors we see. A dog’s sense of smell can be over 10,000 times
as sensitive as ours and is one of the main ways in which they
perceive their environment. Bats and other animals use echo-
location, a sense we do not possess. These senses are central to
an animal’s behavior; for example, they influence how animals
migrate, how they find food, and how they communicate with
one another. Therefore, understanding how animals perceive
their world is key to understanding their behavior.
As you read through the chapters in this book, it will
become clear why understanding behavioral biology is impor-
tant to animal care. However, it can also be helpful to have
a basic understanding of animal behavior as an academic
discipline. Having some knowledge about the way in which
behavioral scientists address behavioral questions can help
guide your own observations. In addition, as with any sci-
entific discipline, there are some terms that are common
in animal behavior that have somewhat different meanings
outside of the field. For example, people often use the terms
“territory” and “home range” interchangeably, but the former
implies active defense, while the latter does not. This distinc-
tion may not seem critically important, but it has implications
for the animal. Vervet monkeys are territorial in nature; rhe-
sus macaques, on the other hand, maintain home ranges but
are not territorial. This seemingly subtle difference is impor-
tant to those responsible for socially housing these species. A
common method of pair housing rhesus macaques involves
moving one potential partner next to the cage of the other
(Truelove et al. 2017). Because vervets are territorial in their
natural environment, this method is not recommended for
them (see Jorgensen 2021).
Our goal for this chapter is to introduce you, the reader, to
the field of animal behavior. In the first section, we describe
steps involved in studying behavior, including biologically rel-
evant questions used by ethologists to explain and understand
behavior. We then discuss some behavioral concepts important
to the care of animals in the laboratory, which are addressed
by the authors in the taxa-specific part of the book. Clearly, a
single chapter cannot cover the entire field of animal behavior.
However, this chapter will, hopefully, provide an overview that
will help put into context the specific information presented in
the other chapters.
The Definition and Challenge
of Animal Behavior
The field of animal behavior, or ethology, broadly described, is
the study of everything that animals are observed or inferred to
do, which includes, but is not limited to, movements, foraging
activities, breeding and parental behavior, predator-prey inter-
actions, communication, migration, learning, cognitive capabil-
ities, and in testing situations, reactions to a variety of stimuli.
Below we describe three phases for the study of animal
behavior.
Description Phase
The study of animal behavior starts first with description, a
basic, but detailed, characterization of the behavior of a spe-
cies. A number of steps are involved in this process. The first
step is to acquire knowledge about the general behavior of
the species in question. Many sources are available online to
assist in this process, including general scientific journals (e.g.,
Animal Behaviour, Institute for Laboratory Animal Welfare
Journal, Journal of Neuroscience, Applied Animal Behaviour
Science), species-specific journals (e.g., American Journal
of Primatology, Mammalogy, Journal of Ornithology), book
chapters (such as those in Part 2 of this volume), monographs,
and online video clips, as well as other reputable sources, such
as The Audubon Society, National Geographic, The Cornell
Lab of Ornithology, the National Centre for the Replacement,
Animal Behavior
Refinement & Reduction of Animals in Research (NC3Rs),
and the National Research Council’s Guide for the Care and
Use of Laboratory Animals.
Because the above information is either in the form of writ-
ten descriptions or short video clips that are selected only to
highlight some activity, the second step involves the live obser-
vation of representative members of the species. Live observa-
tion is crucial in understanding the natural ebb and flow of
behavior over time. Finally, this effort culminates in the devel-
opment of an ethogram. Ethograms describe and define behav-
iors of interest. They can include species-typical behaviors,
as well as abnormal, idiosyncratic behaviors that may not be
displayed much in the animals’ natural habitat. Additionally,
because scientists often use apparatuses and other testing situ-
ations to assess behavior (e.g., Skinner box, open field, radial
arm maze, and Novel Object Test), it is important to add behav-
iors or behavioral constructs derived from these situations to
the ethogram. For example, in the open field test for rodents,
one would include not only locomotion but also proximity to
walls (to measure thigmotaxis, or preferences for walls).
Ethograms ensure that all observers are using a common
language or terminology. While many behaviors are rather
straightforward (e.g., drinking water from a water bottle), oth-
ers may be open to interpretation. For example, you will read
about “motor stereotypies” in the later chapters, which are
often defined as repetitive behaviors, such as pacing and route
tracing. Operationally defining these behaviors is not always
straightforward. If you were to witness a dog walk across his
pen one time, you likely would not consider that behavior to be
pacing. On the other hand, if that same dog were to walk back
and forth, in the same pattern, 50 times, you would probably
(and rightly) call that pacing. But, what if the dog walks back
and forth twice? Would you consider that to be pacing behav-
ior? Effective ethograms clearly define each behavior, reduc-
ing this kind of ambiguity. For example, Jorgensen (2021)
defines pacing as when an animal (a vervet monkey in this
case) walks back and forth at least three times. Many of the
FIGURE 2.1 Example of a rhesus macaque inspecting (a) and avoiding (b) a brightly colored bird toy placed on the home cage as part of a novel
object test. (Republished by permission of Taylor and Francis Group, LLC, a division of Informa plc, from the The Handbook of Primate Behavioral
Management, S.J. Schapiro (Ed), 2017.)
9
chapters in Part 2 of this book contain ethograms for particular
species. Additional ethograms are presented in Part 3.
Quantification Phase
Following the development of an ethogram, careful consider-
ation must be given to how, where, and when the behavioral
categories will be assessed and measured. Actual observa-
tions are at the heart of most behavioral research. Depending
on the circumstance, these observations may be collected
directly by the observer standing in front of the animal, or
more remotely, by an observer scoring videotapes or live web
cams. Additionally, some behaviors can be quantified through
automated systems. For example, jumping and somersaulting
in deer mice can be quantified using a photo beam array, in
which the beams are set high enough so that all four paws must
leave the ground to break the beam (Bechard and Lewis 2016).
Standard measuring units include latencies (i.e., amount of
time between a starting point and the occurrence of a particu-
lar behavior), frequencies (i.e., the number of times a behavior
occurs), and durations (i.e., the length of time).
Observations can occur in many different situations.
Normative or baseline observations often take place either in
an animal’s home cage environment or in unmanipulated free-
ranging populations. Laboratory animals are often studied in
response to some challenge, either using a testing arena (such as
an elevated T-maze) or in their home cage (e.g., Figure 2.1). Free-
ranging animals can also be tested in areas in which manipu-
landa are set out in the field and various capabilities assessed
(Figure 2.2; see van de Waal et al. 2013 for more details).
Because animals may be studied in social groups, issues
of how to observe multiple subjects arise. One can use focal
sampling, in which individuals are observed one at a time for
a certain amount of time (e.g., 15 min a day for several days),
and behaviors of interest are recorded. Conversely, one can use
scan sampling, in which a group of animals is observed, and
the behavior of each individual is recorded at set intervals (e.g.,
10
FIGURE 2.2 Example of testing in free-ranging animals. This study
(van de Waal et al. 2013) examined social learning of food preference in
wild vervet monkeys. Monkeys that immigrate into new troops adopt the
local traditions and eat food even if that food was avoided in their previ-
ous group. (Photo copyright Erica van de Waal.)
one would record the behavior of all animals in a group every
60 s for a 30 min time period). Subject sampling procedures
are determined by the research question (see Altmann 1974).
However, to be effective, observation periods must be long
enough to measure the behavior of interest, and frequent
enough across days and perhaps weeks to ensure that unfore-
seen or uncommon events (e.g., illness, change in care taker,
etc.) do not skew the data. For example, if someone were to
take observations of you at this moment, they would observe
you reading this book (and hopefully learning a great deal
from it!). If that were the only observation period, the observer
might come away thinking that you spend all of your time
reading this book. While we think that would be a great use of
your time, it is probably not the only thing you will do today
or this week. Thus, taking shorter, more frequent observations
over several days, as opposed to longer, less frequent obser-
vations, generally provides a more complete activity budget
of the individual. Detailed descriptions methodologies and
observational sampling techniques are beyond the scope of
this chapter; however, there are very good resources avail-
able on these topics, including Jeanne Altmann’s classic paper,
Observational study of behavior: Sampling methods (Altmann
1974) and Measuring Behavior: An Introductory Guide
(Martin and Bateson 2007).
The Explanation Phase
The field of animal behavior is not merely about description
and quantification. Another important phase is determining
why animals behave the way they do. However, developing
accurate explanations of behavior is extraordinarily compli-
cated. Many factors contribute to the production of behavior.
In fact, the expression of behavior can be considered the ulti-
mate end product of a complex series of interactions between
the organism and the environment.
The sources of these interactions can be divided into two
general types: internal variables (e.g., neurotransmitters,
hormones, gene expression) and external variables (e.g.,
experience, food availability, predation risk, environmental
Behavioral Biology of Laboratory Animals
complexity). Complicating the picture further is the fact that
organisms develop and change over time, which in turn can
yield changes in how they respond to these variables. Thus,
any attempt to explain behavior must take into account the
developmental stage of the organism.
Here we present two examples to illustrate the points raised
above. Meadow voles and prairie voles are closely related rodent
species within the genus, Microtus. However, whereas meadow
voles are promiscuous breeders and females almost exclusively
care for pups, many prairie voles show social monogamy, form-
ing pair bonds with their partners and engaging in bi-parental
care of pups (see reviews, Gobrogge and Wang 2016; Rogers et
al. 2018). One question that has been considered extensively is
why male prairie voles care for pups. Although this process has
been described numerous times and various patterns of behavior
measured (e.g., time spent crouching over pups, number of pup
retrievals), the explanation of why this occurs is considerably
more challenging. Many variables contribute to paternal behav-
ior, including internal factors, such as vasopressin and oxyto-
cin, neuropeptides implicated in social bonding (Bamshad et al.
1994; Bosch and Young 2018; Gobrogge and Wang 2015), and
changes in gene expression (Barrett et al. 2013). Environmental
pressures include mate guarding as a means to ensure paternity
(Getz et al. 2003; Wolff et al. 2002) and changes in population
density (Getz and McGuire 2008). From a developmental per-
spective, the quality of paternal care is dependent, in part, on
previous experience with pups (Stone et al. 2010), allowing ado-
lescent males to perfect basic parenting skills.
In the second example, African Thomson’s gazelles engage
in a rather startling behavior when pursued by a cheetah.
Instead of running as fast as possible in the opposite direction,
they perform an unusual behavior called stotting. The gazelles
run a short distance in the opposite direction and then paradoxi-
cally leap high in the air, run again and leap high in the air, and
continue to do this as long as the cheetah maintains its pursuit.
It is not difficult to describe this phenomenon. In fact, one can
film it and obtain various descriptive measures of average leap
height and number of leaps per unit time. The challenge is to
explain why gazelles stott. Very young gazelles do not stott,
thus there is a developmental component. Additionally, both
external variables (e.g., type of predator, whether the gazelle
is alone or in the middle of a herd) and internal variables (e.g.,
activation of neurotransmitters, increased secretion of adrena-
line) play a role in explaining this behavior. However, the
interactions between these variables are not fully understood.
Furthermore, many other variables remain to be identified.
Biologically Relevant
Questions about Behavior
As demonstrated by the examples above, one of the challenges
facing behavioral scientists is that a variety of factors internal
and external to the organism contribute to the expression of
behavior. Many years ago, Niko Tinbergen, an internation-
ally recognized ethologist, created a structure for explaining
behavior by identifying four biologically relevant questions
(Tinbergen 1963). Two of these were termed proximate ques-
tions, proximate because they depended on understanding
Animal Behavior
behavior in individual animals at the present moment in time.
The remaining two were termed ultimate questions, because
they relied on the process of natural selection, which frequently
required many generations. While most often employed by
behavioral scientists, these questions can help guide the ways
in which we look at the behavior of the animals in our care.
Proximate Questions (Mechanism
and Development)
The first proximate question is concerned with the underly-
ing mechanisms that explain how behavior is expressed. These
include, but are not limited to, the role of neurotransmitters,
hormones, genes, epigenetic processes, neural circuitry, etc.
The second proximate question involves an exploration of the
role of development, and focuses both on the need for critical
internal and external input as young organisms mature, as well
as on the timing of such input.
Ultimate Questions (Function
and Evolutionary History)
The first ultimate question explores the functional significance
of behavior from the standpoint of the ways in which the behav-
ior affects reproductive success. This question also involves
a consideration of the costs and benefits of such behavior, to
explain why it persists in a population. The second question is
concerned with how the behavior may have developed within
various taxonomic groups, and involves a comparison of exist-
ing species using both morphological and molecular tech-
niques. The output of these comparisons is a putative history
of how various species groups are related to one another and
how traits may have arisen over evolutionary time.
Each question formulated by Tinbergen can be viewed as
representing a different, but equally important, level of analy-
sis. Traditionally, scientists focus on only one level of analysis;
e.g., behavioral ecologists look at ultimate causes for behav-
ior, while neuroscientists typically focus on the role of neu-
rotransmitters, etc. However, there may be significant value to
scientific creativity through awareness of findings at all inter-
pretational levels. Similarly, interpreting behavior at multiple
levels can provide a framework for understanding why animals
in our care behave the way they do. Much of the material about
behavior presented in the chapters of this book is provided in
a functional and/or evolutionary context. Authors describe
behaviors that have been shaped by natural selection and pro-
vide information about how to promote those behaviors in the
laboratory environment. Unless you are specifically studying
the underlying neural mechanisms of behavior, you may not
think too much about the proximate mechanisms involved.
However, to really understand why animals are engaging in
behavior, it is important to consider both proximate and ulti-
mate causes. As stated above, experience, particularly early in
life, is one proximate mechanism that underlies behavior of
animals. Early interactions with the mother can influence later
behavior of young animals of a variety of species. For exam-
ple, mouse pups raised by mothers that licked and groomed
them a great deal go on to show a decreased startle response
and increased exploration in an open field compared to pups
11
that received less maternal licking and grooming (Caldji et al.
1998). In addition, certain drugs or scientific treatments can
affect hormones and/or neurotransmitters, which can alter
behavior. Genome editing technologies (such as CRISPR) alter
the genetic code and may have unexpected downstream behav-
ioral effects. As an example, Qiu et al. (2019) used CRISPR/
Cas9 editing to knock out a gene involved in circadian rhythm,
BMAL1, in cynomolgus macaque embryos. As expected, these
monkeys, as juveniles, had altered sleep patterns, but they also
showed decreased exploration and increased fear and anxiety
toward caregivers, compared to control animals. Any time
new technologies that can alter neural function are introduced,
there is the chance that they can also affect the behavior of the
animals. Below, we look at a single example and examine it
from both a proximate and ultimate perspective.
Bird Song as an Example of a Levels
of Analysis Perspective
Some birds produce melodious sounds referred to as song.
Song learning is present only in three distinct avian groups,
constituting somewhat less than 50% of all avian species.
These are the oscine passerine birds, hummingbirds, and par-
rots. Song is distinct from the species-typical calls found in all
avian species (e.g., begging calls, alarm calls, food location
calls), in terms of its structure, complexity, and its sex speci-
ficity (only male birds sing). Although females do not sing,
they nonetheless have to recognize the songs of males of their
species. Bird song has been studied extensively and consider-
able information is available relating to the four biologically
relevant questions (levels of analysis).
1. Mechanism: Although many aspects of bird song
have been explored in this domain, we highlight the
research on the neuroanatomical structures involved
in song (Wild 2004). Birds that sing must first
learn the species song before reliably producing it.
Extensive evidence suggests that these two processes
(learning and producing) are controlled by different
neuroanatomical structures. Pathways that connect
areas of the brain, specifically the lateral magnocel-
lular nucleus of the anterior nidopallium with area X,
appear to be activated during the learning of song.
In contrast, the ongoing production of song involves
a complex circuit that includes the robust nucleus
of the arcopallidum, the high vocal center, and the
hypoglossal nucleus that connects to muscles of the
syrinx or voice box. The distinction between learn-
ing song and producing it is explored further under
the next section “Development”.
2. Development: Male white-crowned sparrows do not
begin to sing until about 200 days of age, and even
then, the song is quite different from the species-
typical song. Extensive research on the white-
crowned sparrow has shown that song learning
is dependent on a combination of innate cues and
crucial experiences across development (Brainard
and Doupe 2002). Without previous song exposure,
young white-crowned sparrows are genetically
12
attuned to respond to conspecific song with increased
heart rate and begging responses (Nelson and Marler
1993). Subsequently, male white-crowned sparrows
acquire the full species song in stages. During the
sensory stage, nestlings hear their father sing the
species song and encode it into their memory. For
maximal memory encoding, exposure should occur
before 100 days of life (Nelson et al. 1997). They then
fledge and begin to sing at around 200 days of age.
Initially, the songs produced are imperfect versions
of the species song. During the sensory-motor stage,
male birds appear to compare their vocal output
with their encoded memory, improving their song
through successive approximations. Males enter the
final stage of crystallization when a match between
the two sources of information is achieved (Nelson
and Marler 1994). At that point, the song remains
relatively invariant throughout the remainder of the
bird’s life, and thus, the white-crowned sparrow
is a closed-end learner. It should be noted that the
process of song learning as described in the white-
crowned sparrow is also observed in zebra finches.
However, it is by no means universal. Many other
bird species are open-ended learners. For example,
canaries retain some old songs from the previous
year, while also learning new songs in the new year.
3. Function: Singing has some very clear costs.
Consider that adult males perch on high branches
and loudly sing their song. What better way to attract
predators. Additionally, singing is energetically
costly and takes time away from foraging for food.
Given those clear costs, the benefits must be pow-
erful enough to exceed those costs. There are obvi-
ously many proposed explanations for bird song that
include, but are not limited to, attracting mates and
defending territories from intruder males.
Here we examine one function, namely, that songs
aid in territorial defense and act to repel intruder
males. Testing the territorial defense hypothesis is
not as easy as it seems. Males not only sing, they also
engage in aggressive behavior against intruders. How
does one separate the behavior from the song? In an
ingenious experiment, this hypothesis was tested by
replacing song sparrow males with speakers playing
their song and examining the rate of territorial intru-
sion (Nowicki et al. 1998). The authors first audiotaped
the songs of 22 males. Then 11 matched pairs of males
were removed temporarily from their territories. One
male of each pair was replaced with a loudspeaker
producing its song whereas the other male’s territory
remained silent. The mean number of intrusions per
hour was significantly lower on territories in which the
male’s song was played, compared to the silent territo-
ries, thus providing support for the idea that song alone
can temporarily deter intrusion by strange males.
4. Evolutionary History: Three groups of birds learn
songs; the oscine passerine birds, hummingbirds,
and parrots. However, the actual ancestral state
Behavioral Biology of Laboratory Animals
reconstruction of this pattern across avian species is
not well established. Initial research involving mor-
phological and molecular comparisons suggested that
these three groups were unrelated and song must have
evolved independently three separate times (Sibley
et al. 1988). A subsequent comparison suggested that
parrots may be closely related to oscine passerine
birds, and thus, song learning may only have evolved
independently twice (Hackett et al. 2008). New infor-
mation now suggests that some suboscine passerine
bird species may also learn their song (Kroodsma
et al. 2013), all of which indicates that the phylogeny
of bird song remains uncertain (Jarvis et al. 2014).
Using the Scientific Method
to Study Behavior
We can generate endless questions about animal behavior and
develop numerous explanations. However, use of the scien-
tific method is essential in determining whether a particular
explanation is a viable answer to a question. For example, Tim
Caro (1986) proposed and tested an explanation for stotting in
gazelles. He started with a simple question; why do gazelles
stott? He then hypothesized that gazelles leap high in the air
to get a better view of the actual location of the cheetah. He
termed this the terrain hypothesis. If the terrain hypothesis
were true, he predicted that the environment would matter,
namely, that in tall grass situations, where cheetah movements
might be difficult to detect, gazelles should stott. In contrast,
in short grass situations, where cheetahs could be easily seen,
stotting would be unnecessary. He then tested this predic-
tion by observing cheetahs and gazelles in both tall and short
grass situations, and discovered that gazelles were equally
likely to stott in both situations. Thus, he ruled out the terrain
hypothesis. Frequently, multiple explanations (hypotheses)
are possible. In some cases, two competing hypotheses that
yield different predictions can be tested simultaneously. Such
hypotheses are considered mutually exclusive.
Caro went on to test four mutually exclusive hypotheses of
stotting in gazelles. Two of these were based on the idea that
stotting was a form of communication to other gazelles, either
to warn them of the presence of a predator or to cause them
to group together to dilute the risk of predation. The remain-
ing two were based on potential interactions with the predator,
either to confuse the predator when many gazelles were stot-
ting at the same time or to alert the predator that it had been
detected and further pursuit was futile (cheetahs rely on stealth
to sneak up on prey). Without going into the details of the pre-
dictions and testing, Caro found support for only one hypoth-
esis related to communication between predator and prey,
namely, detection. However, several years later, Fitzgibbon
and Fanshawe (1988) proposed that gazelles stotted to indi-
cate their fitness to the predator. Unfortunately, detection and
fitness are not mutually exclusive hypotheses, because both
lead to the same prediction. Gazelles should stott to alert the
cheetah that it has been detected, enabling them to outrun the
predator, or they should stott as a demonstration of superior
Animal Behavior
strength and stamina to the cheetah, indicating that they can
outrun the predator. However, a further examination of the
stotting behavior combined with the success or failure of the
hunt provided support for the fitness hypothesis. Gazelles that
were taken by a predator stotted less frequently per unit time
and did not attain the higher leaps of the gazelles that escaped.
The story does not end here. Recent evidence indicates that
females are more likely to stott than males, and stotting may be
used by mothers to warn fawns of dangerous situations (Blank
2018). These varied findings suggest that stotting serves more
than one purpose and that its use is context dependent, a not
uncommon finding in much of animal behavior research.
This example illustrates several points. First, it demon-
strates the rigor with which behavioral science is performed.
It also shows that, as with other fields of science, explanations
may change over time, as new information is uncovered. Much
of the information about the behavior of the species discussed
in this book was learned through rigorous behavioral studies.
Concepts from Animal Behavior That Impact
Laboratory Animal Care and Welfare
There are a number of concepts derived from studying ani-
mals in nature that may have some application to laboratory
animal care and welfare. Both regulatory documents over-
seeing animal care (e.g., United States Animal Welfare Act)
and the Guide for the Care and Use of Laboratory Animals
(National Research Council 2011) emphasize the importance
of promoting species-typical behavior for research animals. In
the sections below, we highlight various evolutionary selec-
tion pressures and features of the environment that act to pro-
duce species-typical behavior. Additionally, we provide some
suggestions for using this information to improve the captive
environment.
Ecology
Spatial Distributions
Animals are distributed across space and time in various
social or nonsocial configurations. Two concepts are used to
characterize spatial distribution: home range and territory.
Home range refers to an area that an animal or group traverses
in their daily search for food, water, mates, resting sites, and
sleeping sites (first defined by Burt 1943). Territories are areas
within the home range that are actively defended from intru-
sion by conspecifics. There are benefits to territoriality for ani-
mals, including exclusive access to food and mates, as well as
safety from predation. However, these benefits come at a cost
(associated with defense), and territoriality may only “make
sense”, if the benefits outweigh the costs. All animals have
home ranges, but not all animals defend territories. Further
complicating the picture is that spatial distributions can vary
across time (breeding vs. nonbreeding season) and within a
species by sex.
A comparison of vole species is useful in characterizing
this variability. Although population-level differences exist, in
general, prairie vole males are socially monogamous, occupy
13
and defend a territory (an area near the nesting burrow), and
help care for the pups. Males show intense aggressive behavior
toward male conspecifics, particularly after mating (Winslow
et al. 1993). Prairie vole home ranges are much larger than
those of meadow voles, and encompass areas of dry grasslands
and hay fields. During the nonbreeding season, prairie voles
are often found in communal groups consisting of the breed-
ing pair and offspring, as well as other males (Getz et al. 1993).
In contrast, promiscuous meadow voles have a different spa-
tial pattern (Pritchett-Corning and Winnicker 2021). Females
tend to exist alone in territories independent of other females,
whereas males have larger home ranges that overlap the ter-
ritories of several females and the home ranges of other males
(Madison 1980). Males and females interact at the time of mat-
ing, and typically only females care for the pups (McGuire and
Novak 1984). During the nonbreeding season, meadow voles
overwinter in larger communal groups, consisting of females
and their offspring and males (Madison and McShea 1987).
In this example, studies of home range and territoriality can
be used to inform the establishment of laboratory breeding
colonies of prairie voles and meadow voles from wild stock.
Whereas prairie voles can ideally be maintained as breeding
pairs, maintaining meadow voles as breeding pairs can be more
difficult, and other options may be necessary. Additionally, the
above findings suggest that sociality may be driven in part by
day length, potentially necessitating light cycle management
in captivity.
Predation and the Landscape of Fear
Another factor that can influence habitat use is the risk of
predation. Predation risk in free-ranging animals is known to
vary across different environments, presumably eliciting dif-
ferent perceptions of risk. This spatial variation in the percep-
tion of risk by prey species is referred to as the landscape of
fear (Laundré 2010). As risk increases, prey species respond
with increased vigilance behavior and with changes in habi-
tat use (Blanchard et al. 2018). For example, a shift in habitat
preference from aspen stands to conifer forests was observed
in elk after the reintroduction of wolves to Yellowstone Park
(Fortin et al. 2005).
Clearly, changing habitats is not the only way that animals
respond to predation threat. There are many ways in which
animals may respond to a potential predator. One common
response is to remain motionless, or freeze, to avoid detection;
some animals, such as various frog species (e.g., Ramalho et al.
2019), even feign death by adopting a rigid posture in addition
to the immobility (see Figure 2.3). Many animals flee and/
or hide when detected by a predator, while others fight back,
with chemical (e.g., skunk spray) or mechanical (e.g., teeth)
defenses. Individuals do not always have to avoid predators
on their own; individuals within a group may work together to
“mob” a potential predator. The chapters in Part 2 of this book
describe anti-predator behaviors for the various species.
Do such animals perceive the laboratory environment as
a landscape of fear? Obviously, laboratory animals are not
exposed to predators or predator scent, unless such exposure is
part of a research protocol (or, if carestaff inadvertently intro-
duce predator scent by, for example, working with a predator
14
FIGURE 2.3 South African mongoose (left) feigns death (right) when approached by a human. (Photos by M. Novak.)
species, such as a cat, before working with a prey species, such
as a mouse). However, it is reasonable to consider that species-
typical reactions to predators honed over evolutionary time
by natural selection can impact behavior in the laboratory.
For example, research tamarins (small New World primates,
closely related to the marmosets discussed in Chapter 24)
that lived in a laboratory environment their entire lives, were
given a choice of sleeping boxes with differing levels of con-
cealment. The tamarins preferred the boxes with the highest
level of cover, even though they had never seen a predator.
Further, when forced to sleep in boxes with relatively little
concealment, they increased their rates of vigilant scanning
(Caine et al. 1992). They maintained this antipredator behav-
ior, despite the lack of predators in their environment.
There are, of course, events that occur across the day that
can increase the perception of risk for laboratory animals.
Human observers and/or caretakers may be viewed as poten-
tial threats (Caine 2017, Waiblinger et al. 2006), particularly
if they are involved in a negative interaction (e.g., handling
for injections). There are other events that can be perceived
as high risk, including being transferred to new rooms, new
locations, or new partners. It is likely that some of the natural
fear responses (increased vigilance and movements indicative
of flight) can be mitigated by desensitization and positive rein-
forcement training techniques (e.g., blood glucose testing in
chimpanzees: Reamer et al. 2014; reducing abnormal behav-
ior: Baker et al. 2009; Coleman and Maier 2010) and by pro-
viding safe havens when possible (e.g., transferring marmosets
to new cages by carrying them in their nest boxes).
Social Interactions and Organization
Social Organization
Social organization refers to the structural features of social
relationships that include, but are not limited to, sexual com-
position, age composition, cohesiveness, division of labor,
and patterns of communication. Social organization varies
widely, ranging from solitary females, whose primary social
interaction is with offspring, to large multimale, multifemale
Behavioral Biology of Laboratory Animals
social groups consisting of every age class (infants, juveniles,
adolescents, and adults). An example of such widespread vari-
ability can be observed in the primate order. Five distinct
organizations are recognized: (1) Solitary living: Only a very
few primate species are solitary (e.g., lorises, galagos, orang-
utans), with the social unit being the mother-infant dyad. In
this form of social organization, solitary males form overlap-
ping home ranges around solitary females. (2) Monogamy:
Relatively few primate species live as breeding pairs sur-
rounded by immature offspring that disperse prior to sexual
maturity (e.g., gibbons, siamangs, owl monkeys, titi mon-
keys). These species tend to be territorial and monomorphic.
(3) Polyandry: In polyandrous groups, one resident female
lives with several males. This social structure is also quite
rare, but has been observed in some tamarins. (4) Polygyny:
In polygynous groups, one resident male lives with several
females (e.g., gelada and hamadryas baboons, langurs, patas
monkeys) in a “harem” group. Coexisting with these harem
groups are all-male groups. Males from these all-male groups
frequently attempt to oust harem males, and the tenure of
individual harem males is often short. (5) Polygynandry:
Polygynandrous groups consist of multiple adult males and
multiple adult females plus offspring of varying ages. This is
a common form of primate social structure typical of many
macaques, most baboon species, vervet monkeys, capuchins,
ringtailed lemurs, and some colobines.
The problem with creating the above classification is that
within-species variation is not adequately represented. For
example, marmosets are considered monogamous primates
that live in extended family groups, and most studies of free-
ranging marmosets are consistent with this view (Arruda et al.
2005; Yamamoto et al. 2014). However, other marmoset social
groupings have been reported, including multimale and mul-
tifemale groups of individuals that are not necessarily closely
related (Faulkes et al. 2003), as well as groups containing two
breeding females (Digby and Ferrari 1994). Marmosets are not
the only primate species to show such variability. There are
other factors, such as resource availability and predation pres-
sure, that are not taken into account in the above scheme, yet
do influence social groupings.
Another random document with
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the years rolled on, the bitterness of those memories grew less bitter
and still the sweet remained.
Presently, as for the hundredth time the beloved name met my
eye, there came upon me a yearning to creep back with her into the
sunny past; to forget, if only for a short hour, the hideous anxieties of
the present and in memory to walk with her once more “along the
meads of asphodel.”
Halting before the box, I stood and lifted it tenderly to the table
and having unlocked it, raised the lid and looked thoughtfully into the
interior. Then, one by one, I lifted out my treasures, set them out in
order on the table and sat down to look at them and let them speak
to me their message of peace and consolation.
To a stranger’s eye they were a mere collection of odds and
ends. Some would have been recognizable as relics of the more
conventional type. There were several photographs of the dead girl,
some taken by myself, and a tress of red-gold hair—such hair as I
had been told often glorifies the victims whom consumption had
marked for its own. It had been cut off for me by Barbara when she
took her own tress, and tied up with a blue ribbon. But it was not
these orthodox relics that spoke to me most intimately. I had no need
of their aid to call up the vision of her person. The things that set my
memory working were the records of actions and experiences; the
sketch-books, the loose sketches and the little plaster plaques and
medallions that she had made with my help after she had become
bed-ridden and could go no more abroad to sketch. Every one of
these had its story to tell, its vision to call up.
I turned over the sketches—simple but careful pencil drawings for
the most part, for Stella, like me, had more feeling for form than for
colour—and recalled the making of them; the delightful rambles
across the sunny meadows or through the cool woodlands, the
solemn planting of sketching-stools and earnest consultation on the
selection and composition of the subjects. These were the happiest
days, before the chilly hand of the destroyer had been laid on its
chosen victim and there was still a long and sunny future to be
vaguely envisaged.
And then I turned to the little plaques and medallions which she
had modelled and under my supervision and of which I had made
the plaster moulds and casts. These called up sadder memories, but
yet they spoke of an even closer and more loving companionship; for
each work was, in a way, a joint achievement over which we had
triumphed and rejoiced together. So it happened that, although the
shadow of sickness, and at last of death, brooded over them, it was
on these relics that I tended to linger most lovingly.
Here was the slate that I had got for her to stick the clay on and
which she used to hold propped up against her knees as she worked
with never-failing enthusiasm through the long, monotonous days,
and even, when she was well enough, far into the night by the light
of the shaded candle. Here were the simple modelling-tools and the
little sponge and the Camel-hair brush with which she loved to put
the final finish on the damp clay reliefs. Here was Lanterri’s priceless
text-book over which we used to pore together and laud that
incomparable teacher. Here were the plaques, medals and
medallions that we had prised out, with bated breath, from their too-
adherent moulds. And here—the last and saddest relic—was the
wax mould from which no cast had ever been made, the final,
crowning work of those deft, sensitive fingers.
For the thousandth time, I picked it up and let the light fall
obliquely across its hollows. The work was a medal some three
inches across, a portrait of Stella, herself, modelled from a profile
photograph that I had taken for the purpose. It was an excellent
likeness and unquestionably the best piece of modelling that she had
ever done.
Often, I had intended to take the cast from it, but always had
been restrained by a vague reluctance to disturb the mould. Now, as
I looked at the delicate, sunken impression, I had again the feeling
that this, her last work, ought to be finished; and I was still debating
the matter with the mould in my hand when I heard a quick step
upon the stair, followed by a characteristic knock on my door.
My first impulse was to hustle my treasures back into their box
before answering the summons. But this was almost instantly
followed by a revulsion. I recognized the knock as Thorndyke’s; and
somehow there came upon me a desire to share my memories with
him. He had shown a strangely sympathetic insight into my feelings
towards Stella. He had read my diary. He now knew the whole story;
and he was the kindest, the most loyal and most discreet of friends.
Gently laying down the mould I went to the door and threw it open.
“I saw your light burning as I passed just now,” said Thorndyke as
he entered and shook my hand warmly, “so I thought I would take
the opportunity to drop in and return your diary. I hope I am not
disturbing you. If I am, you must treat me as a friend and eject me.”
“Not at all, Thorndyke,” I replied. “On the contrary, you would be
doing me a charity if you would stay and smoke a companionable
pipe.”
“Good,” said he, “then I will give myself the pleasure of a quiet
gossip. But what is amiss, Mayfield?” he continued, laying a friendly
hand on my shoulder and looking me over critically. “You look worn,
and worried and depressed. You are not letting your mind dwell too
much, I hope, on the tragedy that has come unbidden into your life?”
“I am afraid I am,” I replied. “The horrible affair haunts me.
Suspicion and mystery are in the very air I breathe. A constant
menace seems to hang over all my friends, so that I am in continual
dread of some new catastrophe. I have just ascertained that
Wallingford is really being watched and shadowed; and not only
Wallingford but even Miss Norris.”
He did not appear surprised or seek for further information. He
merely nodded and looked into my face with grave sympathy.
“Put it away, Mayfield,” said he. “That is my counsel to you. Try to
forget it. You have put the investigation into my hands. Leave it there
and wash your own of it. You did not kill Harold Monkhouse.
Whoever did must pay the penalty if ever the crime should be
brought home to the perpetrator. And if it never can be, it were better
that you and all of us should let it sink into oblivion rather than allow
it to remain to poison the lives of innocent persons. Let us forget it
now. I see you were trying to.”
I had noticed that when he first entered the room, he cast a
single, swift glance at the table which, I was sure, had
comprehended every object on it. Then he had looked away and
never again let his eyes stray in that direction. But now, as he
finished speaking, he glanced once more at the table, and this time
with undisguised interest.
 “Yes,” I admitted. “I was trying to find in the memories of the past
an antidote for the present. These are the relics of that past. I
daresay you have read of them in the diary and probably have
written me down a mawkish sentimentalist.”
“I pray you, my friend, not to do me that injustice!” he exclaimed.
“Faithful friendship that even survives the grave, is not a thing that
any man can afford to despise. But for the disaster of untimely death,
your faithfulness and hers would have created for two persons the
perfect life. I assure you, Mayfield, that I have been deeply moved by
the story of your delightful friendship and your irreparable loss. But
don’t let us dwell too much on the sad aspects of the story. Show me
your relics. I see some very charming little plaques among them.”
He picked up one with reassuring daintiness of touch and
examined it through a reading-glass that I handed to him.
“It really is a most admirable little work,” said he. “Not in the least
amateurish. She had the makings of a first-class medallist; the
appreciation of the essential qualities of a miniature relief. And she
had a fine feeling for composition and spacing.”
Deeply gratified by his appreciation and a little surprised by his
evident knowledge of the medallist’s art, I presented the little works,
one after another, and we discussed their merits with the keenest
interest. Presently he asked:
“Has it never occurred to you, Mayfield, that these charming little
works ought to be finished?”
“Finished?” I repeated. “But, aren’t they finished?”
“Certainly not. They are only in the plaster. But a plaster cast is
an intermediate form, just a mere working model. It is due to the
merits of these plaques and medals that they should be put into
permanent material—silver or copper or bronze. I’ll tell you what,
Mayfield,” he continued, enthusiastically. “You shall let Polton make
replicas of some of them—he could do it with perfect safety to the
originals. Then we could hand the casts to an electrotyper or a
founder—I should favour the electrotype process for such small
works—and have them executed in whichever metal you preferred.
Then you would be able to see, for the first time, the real quality of
the modelling.”
I caught eagerly at the idea, but yet I was a little nervous.
 “You think it would be perfectly safe?” I asked.
“Absolutely safe. Polton would make gelatine moulds which
couldn’t possibly injure the originals.”
That decided me. I fell in with the suggestion enthusiastically, and
forthwith we began an anxious consultation as to the most suitable
pieces with which to make a beginning. We had selected half a
dozen casts when my glance fell on the wax mould. That was
Stella’s masterpiece and it certainly ought to be finished; but I was
loath to part with the mould for fear of an accident. Very dubiously, I
handed it to Thorndyke and asked:
“What do you think of this? Could it be cast without any risk of
breaking it?”
He laid the mould on the table before him so that the light fell
obliquely across it and looked down on it reflectively.
“So,” said he, “this is the wax mould. I was reading about it only
yesterday and admiring your resourcefulness and ingenuity. I must
read the entry again with the actual object before me.”
He opened the diary, which he had laid on the table, and when he
had found the entry, read it to himself in an undertone.
“Dropped in to have tea with Stella and found her bubbling with
excitement and triumph. She had just finished the portrait medal and
though her eyes were red and painful from the strain of the close
work, in spite of her new spectacles, she was quite happy and as
proud as a little peacock. And well she might be. I should like
Lanterri to see his unknown pupil’s work. We decided to make the
mould of it at once, but when I got out the plaster tin, I found it
empty. Most unfortunate, for the clay was beginning to dry and I
didn’t dare to damp it. But something had to be done to protect it.
Suddenly I had a brilliant idea. There was nearly a whole candle in
Stella’s candlestick, quite enough for a mould, and good, hard wax
that wouldn’t warp. I took off the reflector and lighted the candle,
which I took out of the candlestick and held almost upside down over
the clay medal and let the wax drip on to it. Soon the medal was
covered by a film of wax which grew thicker and thicker, until, by the
time I had used up practically the whole of the candle, there was a
good, solid crust of wax, quite strong enough to cast from. When I
went home, I took the slate with me with the wax mould sticking to it,
intending to cover it with a plaster shell for extra safety. But my
plaster tin was empty, too, so I put the slate away in a safe place
until I should get some fresh plaster to make the cast; which will not
happen until I get back from Chelmsford.
“Busy evening getting ready for to-morrow; hope I shall feel less
cheap then than I do now.”
As Thorndyke finished reading he looked up and remarked: “That
was an excellent plan of yours. I have seen Polton use the same
method. But how was it that you never made the cast?”
“I was afraid of damaging the mould. As you know, when I came
back from Ipswich, Stella was dead, and as the medal was her last
work and her best, I hardly dared to risk the chance of destroying it.”
“Still,” Thorndyke urged, “it was the medal that was her work. The
mould was your own; and the medal exists only potentially in the
mould. It will come into actual existence only when the cast is made.”
I saw the force of this, but I was still a little uneasy, and said so.
“There is no occasion,” said he. “The mould is amply strong
enough to cast from. It might possibly break in separating the cast,
but that would be of no consequence, as you would then have the
cast, which would be the medal, itself. And it could then be put into
bronze or silver.”
“Very well,” I said, “if you guarantee the safety of the operation, I
am satisfied. I should love to see it in silver; or perhaps it might look
even better in gold.”
Having disposed of the works, themselves, we fell to discussing
the question of suitable settings or frames; and this led us to the
subject of the portraits. Thorndyke glanced over the collection, and
picking up one, which happened to be my own favourite, looked at it
thoughtfully.
“It is a beautiful face,” said he, “and this seems to have been a
singularly happy portrait. In red chalk autotype, it would make a
charming little picture. Did you take it?”
“Yes; and as I have the negative I am inclined to adopt your
suggestion. I am surprised that I never thought of it myself, for red
chalk is exactly the right medium.”
“Then let Polton have the negative. He is quite an expert in
autotype work.”
 I accepted the offer gladly and we then came back to the
question of framing. Thorndyke’s suggestion was that the portrait
should be treated as a medallion and enclosed in a frame to match
that of the medal. The idea appealed to me rather strongly, and
presently a further one occurred to me, though it was suggested
indirectly by Thorndyke, who had taken up the tress of Stella’s hair
and was looking at it admiringly as he drew it softly between his
fingers.
“Human hair,” he remarked, “and particularly a woman’s hair, is
always a beautiful material, no matter what its colour may be; but
this red-gold variety is one of the most gorgeous of Nature’s
productions.”
“Yes,” I agreed, “it is extremely decorative. Barbara had her tress
made up into a thin plait and worked into the frame of a miniature of
Stella. I liked the idea, but somehow the effect is not so very
pleasing. But it is an oblong frame.”
“I don’t think,” said Thorndyke, “that a plait was quite the best
form. A little cable would look better, especially for a medallion
portrait; indeed I think that if you had a plain square black frame with
a circular opening, a little golden cable, carried round concentrically
with the opening would have a rather fine effect.”
“So it would,” I exclaimed. “I think it would look charming. I had
no idea, Thorndyke, that you were a designer. Do you think Polton
could make the cable?”
“Polton,” he replied, impressively, “can do anything that can be
done with a single pair of human hands. Let him have the hair, and
he will make the cable and the frame, too; and he will see that the
glass cover is an airtight fit—for, of course, the cable would have to
be under the glass.”
To this also I agreed with a readiness that surprised myself. And
yet it was not surprising. Hitherto I had been accustomed secretly
and in solitude to pore over these pathetic little relics of happier days
and lock up my sorrows and my sense of bereavement in my own
breast. Now, for the first time, I had a confidant who shared the
knowledge of my shattered hopes and vanished happiness; and so
whole heartedly, with such delicate sympathy and perfect
understanding had Thorndyke entered into the story of my troubled
life that I found in his companionship not only a relief from my old
self-repression but a sort of subdued happiness. Almost cheerfully I
fetched an empty cigar-box and a supply of cotton wool and tissue
paper and helped him tenderly and delicately to pack my treasures
for their first exodus from under my roof. And it was with only a faint
twinge of regret that I saw him, at length, depart with the box under
his arm.
“You needn’t be uneasy, Mayfield,” he said, pausing on the stairs
to look back. “Nothing will be injured; and as soon as the casting is
successfully carried through, I shall drop a note in your letter-box to
set your mind at rest. Good night.”
I watched him as he descended the stairs, and listened to his
quick foot-falls, fading away up the court. Then I went back to my
room with a faint sense of desolation to re-pack the depleted deed-
box and thereafter to betake myself to bed.
 Chapter XV.
A Pursuit and a Discovery
More than a week had passed since that eventful evening—how
eventful I did not then realize—when I had delivered my simple
treasures into Thorndyke’s hands. But I was not uneasy; for, within
twenty-four hours, I had found in my letter-box the promised note,
assuring me that the preliminary operations had been safely carried
through and that nothing had been damaged. Nor was I impatient. I
realized that Polton had other work than mine on hand and that there
was a good deal to do. Moreover, a little rush of business had kept
me employed and helped me to follow Thorndyke’s counsel and
forget, as well as I could, the shadow of mystery and peril that hung
over my friends, and, by implication, over me.
But on the evening of which I am now speaking I was free. I had
cleared off the last of the day’s work, and, after dining reposefully at
my club, found myself with an hour or two to spare before bed-time;
and it occurred to me to look in on Thorndyke to smoke a friendly
pipe and perchance get a glimpse of the works in progress.
I entered the Temple from the west, and, threading my way
through the familiar labyrinth, crossed Tanfield Court, and passing
down the narrow alley at its eastern side, came out into King’s Bench
Walk. I crossed the Walk at once and was sauntering down the
pavement towards Thorndyke’s house when I noticed a large, closed
car drawn up at its entry, and, standing on the pavement by the car,
a tall man whom I recognized by the lamp light as Mr.
Superintendent Miller.
Now I did not much want to meet the superintendent, and in any
case it was pretty clear to me that my visit to Thorndyke was not very
opportune. The presence of Miller suggested business, and the size
of the car suggested other visitors. Accordingly I slowed down and
was about to turn back when my eye caught another phenomenon.
In the entry next to Thorndyke’s a man was standing, well back in
the shadow, but not so far that he could not get a view of the car; on
which he was quite obviously keeping a watchful eye. Indeed, he
was so pre-occupied with his observation of it that he had not
noticed my approach, his back being turned towards me.
Naturally, the watchful attitude and the object of his watchfulness
aroused my suspicions as to his identity. But a movement backward
on his part which brought him within range of the entry lamp, settled
the matter. He was Anthony Wallingford.
I turned and walked quietly back a few paces. What was this idiot
doing here within a few yards of Thorndyke’s threshold? Was he
merely spying fatuously and without purpose? Or was it possible that
he might be up to some kind of mischief? As I framed the question
my steps brought me opposite another entry. The Walk was in
darkness save for the few lamps and the place was practically
deserted. After a moment’s reflection, I stepped into the entry and
decided thence to keep a watch upon the watcher.
I had not long to wait. Hardly had I taken up my rather undignified
position when three men emerged from the house and walked slowly
to the car. By the light of the lamp above Thorndyke’s entry, I could
see them quite plainly and I recognized them all. One was
Thorndyke, himself, another was Dr. Jervis, Thorndyke’s colleague,
now in the employ of the Home Office, and the third was Dr.
Barnwell, well-known to me as the analyst and toxicologist to the
Home Office. All three carried substantial bags and Dr. Barnwell was
encumbered with a large case, like an out-size suit-case, suggestive
of chemical apparatus. While they were depositing themselves and
their impedimenta in the car, Superintendent Miller gave directions to
the driver. He spoke in clear, audible tones, but though (I have to
confess) I listened intently, I caught only the question: “Do you know
the way?” The words which preceded and followed it were just
audible but not intelligible to me. It appeared, however, that they
were intelligible to Wallingford, for, as soon as they were spoken and
while the superintendent still held the open door of the car, he
stepped forth from his lurking-place and walked boldly and rapidly
across to the narrow passage by which I had come.
 Realizing instantly what his intention was, I came out of the entry
and started in pursuit. As I reached the entrance to the passage, my
ear caught the already faint sound of his receding footsteps; by
which I learned that he was running swiftly and as silently as he
could. Since I did not intend to lose him, I had no choice but to follow
his example, and I raced across Tanfield Court, past the Cloisters
and round by the church as if the Devil were after me instead of
before. Half-way up Inner Temple Lane he slowed down to a walk—
very wisely, for otherwise the night porter would certainly have
stopped him—and was duly let out into Fleet Street, whither I
followed him at a short interval.
When I stepped out of the gate I saw him some little distance
away to the west, giving directions to the driver of a taxi. I looked
round desperately, and, to my intense relief, perceived an apparently
empty taxi approaching from the east. I walked quickly towards it,
signalling as I went, and the driver at once drew in to the kerb and
stopped. I approached him, and, leaning forward, said in a low voice
—though there was no one within earshot:
“There is a taxi just in front. It will probably follow a big car which
is coming up Middle Temple Lane. I want you to keep that taxi in
sight, wherever it may go. Do you understand?”
The man broke into a cynical grin—the nearest approach to
geniality of which a taxi-driver is capable—and replied that he
understood; and as, at this moment, the nose of the car appeared
coming through the arched entrance gate of Middle Temple Lane, I
sprang into the taxi and shut the door. From the off-side window, but
keeping well back out of sight, I saw the car creep across Fleet
Street, turn eastward and then sweep round into Chancery Lane.
Almost immediately, Wallingford’s taxi moved off and followed; and
then, after a short interval, my own vehicle started, and, crossing
directly to Chancery Lane, went ahead in the wake of the others.
It was an absurd affair. Now that the pursuit was started and its
conduct delegated for the time to the driver, I leaned back in the
shadow and was disposed to grin a little sheepishly at my own
proceedings. I had embarked on them in obedience to a sudden
impulse without reflection—for which, indeed, there had been no
time. But was there anything to justify me in keeping this watch on
Wallingford? I debated the question at some length and finally
decided that, although he was probably only playing the fool, still it
was proper that I should see what he was really up to. Thorndyke
was my friend and it was only right that I should stand between him
and any possible danger. Well as he was able to take care of
himself, he could not be always on his guard. And I could not forget
the infernal machine. Some one at least had the will to do him an
injury.
But what about the brown-hatted man? Why had he not joined in
this novel sport? Or had he? I put my head out of the window and
looked along the street in our rear, but there was no sign of any
pursuing taxi. The ridiculous procession was limited to three
vehicles; which was just as well, since we did not want a police
cyclist bringing up the rear.
From my own proceedings my thoughts turned to those of
Thorndyke and his companions, though they were no affair of mine,
or of Wallingford’s either, for that matter. Apparently the three men
were going somewhere to make a post mortem examination. The
presence of Dr. Barnwell suggested an analysis in addition; and the
presence of Miller hinted at a criminal case of some kind. But it was
not my case or Wallingford’s. For both of us the analyst had already
done his worst.
While I reflected, I kept an eye on the passing landmarks,
checking our route and idly trying to forecast our destination. From
Chancery Lane we crossed Holborn and entered Gray’s Inn Road, at
the bottom of which we swept round by King’s Cross into Pancras
Road. At the end of this we turned up Great College Street, crossed
Camden Road and presently passed along the Kentish Town Road.
So far I had noted our progress with no more than a languid interest.
It did not matter to me whither we were going. But when, at the Bull
and Gate, we swept round into Highgate Road, my attention awoke;
and when the taxi turned sharply at the Duke of St. Albans and
entered Swain’s Lane, I sat up with a start. In a moment of sudden
enlightenment, I realized what our destination must be; and the
realization came upon me with the effect of a palpable blow. This
lane, with its precipitous ascent at the upper end, was no ordinary
thoroughfare. It was little more than an approach to the great
cemetery whose crowded areas extended on either side of it; its
traffic was almost completely limited to the mournful processions that
crept up to the wide gates by the mortuary chapel. Indeed, on the
very last occasion when I had ridden up this lane, my conveyance
had been the mourning carriage which followed poor little Stella to
her last home.
Before I had recovered from the shock of this discovery
sufficiently to consider what it might mean, the taxi came to a sudden
halt. I stepped out, and, looking up the lane, made out the shadowy
form of Wallingford’s vehicle, already backing and manœuvring to
turn round.
“Bloke in front has got out,” my driver announced in a hoarse
whisper, and as he spoke, I caught sight of Wallingford—or at least
of a human figure—lurking in the shadow of the trees by the railings
on the right-hand side of the road. I paid off my driver (who,
thereupon, backed on to the footway, turned and retired down the
hill) and having waited for the other taxi to pass down, began slowly
to ascend the lane, keeping in the shadow of the trees. Now that the
two taxis were gone, Wallingford and I had the lane to ourselves,
excepting where, in the distance ahead, the reflected light from the
head-lamps of the car made a dim halo and the shape of the gothic
chapel loomed indistinctly against the murky sky. I could see him
quite plainly, and no doubt he was aware of my presence; at any
rate, I did not propose to attempt any concealment, so far as he was
concerned. His movements had ceased to be of any interest to me.
My entire concern was with the party ahead and with the question at
to what Thorndyke was doing at this time of night in Highgate
Cemetery.
The burial ground is divided, as I have said, into two parts, which
lie on either side of the lane; the old cemetery with its great gates
and the large mortuary chapel, on the left or west side and the newer
part on the right. To which of these two parts was Thorndyke bound?
That was the question that I had to settle.
I continued to advance up the lane, keeping in the shadow,
though it was a dark night and the precaution was hardly necessary.
Presently I overtook Wallingford and passed him without either
concealment or recognition on either side. I could now clearly make
out the gable and pinnacles of the chapel and saw the car turn in the
wide sweep and then extinguish its headlights. Presently, from the
gate-house there emerged a party of men of whom some carried
lanterns, by the light of which I could recognize Thorndyke and his
three companions; and I noted that they appeared to have left their
cases either in the car or elsewhere for they now carried nothing.
They lingered for a minute or two at the wicket by the great gates;
then, accompanied by a man whom I took to be the gate-keeper,
they crossed the road to the gate of the eastern cemetery and were
at once followed by another party of men, who trundled two wheel-
barrows, loaded with some bulky objects the nature of which I could
not make out. I watched them with growing anxiety and suspicion as
they passed in at the gate; and when they had all entered and
moved away along the main path, I came forth from the shadow and
began to walk quickly up the lane.
The eastern cemetery adjoins Waterlow Park, from which it is
separated by a low wall surmounted by tall railings, and this was my
objective. The park was now, of course, closed for the night, locked
up and deserted. So much the better. Locks and bars were no
hindrance to me. I knew the neighbourhood of old. Every foot of the
lane was familiar to me, though the houses that had grown up at the
lower end had changed its aspect from that which I remembered
when as a boy I had rambled through its leafy shades. On I strode,
past the great gates on the left and the waiting car, within which I
could see the driver dozing, past the white gatehouse on the right,
up the steep hill until I came to the place where a tall oak fence
encloses the park from the lane. Here I halted and took off my
overcoat, for the six-foot fence is guarded at the top by a row of
vicious hooks. Laying the folded overcoat across the top of the
fence, I sprang up, sat for a moment astride and then dropped down
into the enclosure.
I now stood in a sort of dry ditch between the fence and a steep
bank, covered with bushes which rose to the level of the park. I had
just taken down my overcoat and was putting it on before climbing
the bank when its place was taken by another overcoat cast over
from without. Then a pair of hands appeared, followed by the clatter
of feet against the fence and the next moment I saw Wallingford
astride of the top and looking down at me.
I still affected to be unaware of him, and, turning away, began to
scramble up the bank, at the summit of which I pushed my way
through the bushes, and, stepping over a three-foot fence, came out
upon a by-path overshadowed by trees. Pausing for a moment to get
my bearings and to mark out a route by which I could cross the park
without coming into the open, where I might be seen by some
watchful keeper, I started off towards a belt of trees just as
Wallingford stepped over the dwarf fence and came out upon the
path behind me.
The position was becoming absurd, though I was too agitated to
appreciate its humour. I could not protest against his following me
seeing that I had come in the first place to spy upon him, and was
now, like himself, engaged in spying upon Thorndyke. However, he
soon solved the difficulty by quickening his pace and overtaking me,
when he asked in a quite matter-of-fact tone:
“What is Thorndyke up to, Mayfield?”
“That is what I want to find out,” I replied.
“He is not acting on your instructions, then?”
“No; and the probability is that what he is doing is no concern of
mine or of yours either. But I don’t know; and I have come here to
make sure. Keep in the shadow. We don’t want the keeper to see us
prowling about here.”
He stepped back into the shade and we pursued our way in
silence; and even then, troubled and agitated as I was, I noted that
he asked me no question as to what was in my mind. He was leaving
the initiative entirely to me.
When we had crossed the park in the shelter of the trees and
descended into the hollow by the little lake where we were out of
sight of the gate-house, I led the way towards the boundary between
the park and the cemetery. The two enclosures were separated, as I
have said, by a low wall surmounted by a range of high, massive
railings; and the wall and the cemetery beyond were partially
concealed by an irregular hedge of large bushes. Pushing through
the bushes, I moved along the wall until I came to the place which I
intended to watch; and here I halted in the shade of a tall mass of
bushes, and resting my arms on the broad coping of the wall, took
up my post of observation with Wallingford, silently attentive at my
side.
The great burial ground was enveloped in darkness so profound
that the crowded headstones and monuments conveyed to the eye
no more than a confused glimmer of ghostly pallor that was barely
distinguishable from the general obscurity. One monument only
could be separately identified: a solitary stone cross that rose above
a half-seen grave some sixty yards from the wall. But already the
mysterious procession could be seen threading its way in and out by
the intricate, winding paths, the gleam of the lanterns lighting up now
a marble figure and now a staring head-stone or urn or broken
column; and as it drew ever nearer, the glare of the lanterns, the
rumble of the barrow-wheels on the hard paths and the spectral
figures of the men grew more and more distinct. And still Wallingford
watched and spoke never a word.
At length, a turn of the path brought the procession into full view,
and as it approached I could make out a man,—evidently by his
uniform, the cemetery keeper,—leading, lantern in hand and showing
the way. Nearer and nearer the procession drew until at last, close
by the stone cross, the leader halted. Then, as Thorndyke and his
companions—now clearly visible—came up, he lifted his lantern and
let its light fall full on the cross. And even at this distance I could read
with ease—though it was unnecessary—the single name STELLA.
As that name—to me so sacred—flashed out of the darkness,
Wallingford gripped my arm. “Great God!” he exclaimed. “It is Stella
Keene’s grave! I came here once with Barbara to plant flowers on it.”
He paused, breathing hard and still clutching my arm. Then, in a
hoarse whisper, he demanded:
“What can that devil be going to do?”
There was little need to ask. Even as he spoke, the labourers
began to unload from the first barrow its lading of picks, shovels and
coils of rope. And when these were laid on the ground, the second
barrow yielded up its cargo; a set of rough canvas screens which the
men began to set up around the grave. And even as the screens
were being erected, another lantern slowly approaching along the
path, revealed two men carrying a long, bedstead-like object—a bier
—which they at length set down upon its stunted legs just outside
the screens.
With set teeth I stared incredulously between the railings at these
awful preparations while Wallingford, breathing noisily, held fast to
my arm with a hand that I could feel shaking violently. The lanterns
inside the screens threw a weird, uncertain light on the canvas, and
monstrous, distorted shadows moved to and fro. Presently, amidst
these flitting, spectral shapes, appeared one like an enormous
gnome, huge, hideous and deformed, holding an up-raised pick. The
shadowy implement fell with an audible impact, followed by the ring
of a shovel.
At the sight and the sound—so dreadfully conclusive—
Wallingford sprang up with a stifled cry.
“God Almighty! That devil is going to dig her up!”
He stood motionless and rigid for a few moments. Then, turning
suddenly, without another word, he burst through the bushes, and I
heard him racing madly across the park.
I had half a mind to follow him. I had seen enough. I now knew
the shocking truth. Why stay and let my soul be harrowed by the
sight of these ghouls. Every stroke of pick or shovel seemed to
knock at my heart. Why not go and leave them to their work of
desecration? But I could not go. I could not tear myself away. There
was the empty bier. Presently she would be lying on it. I could not go
until I had seen her borne away.
So I stayed there gazing between the railings, watching the elfin
shapes that flitted to and fro on the screen, listening to the thud of
pick and the ring and scrape of shovel and letting my confused
thoughts wander obscurely through a maze of half-realized pain and
anger. I try in vain to recall clearly what was my state of mind. Out of
the confusion and bewilderment little emerges but a dull indignation
and especially a feeling of surprised resentment against Thorndyke.
The horrible business went on methodically. By degrees a
shadowy mound grew up at the bottom of the screen. And then other
movements and other sounds; a hollow, woody sound that seemed
to bring my heart into my mouth. At last, the screens were opened at
the end and then the coffin was borne out and laid on the bier. By the
light of the lanterns I could see it distinctly. I was even able to
recognize it, shabby and earth-stained as it now was. I saw
Thorndyke help the keeper to spread over it some kind of pall, and
then two men stepped between the handles of the bier, stooped and
picked it up; and then the grim procession re-formed and began
slowly to move away.
I watched it until it had passed round a turn of the path and was
hidden from my view. Then I stood up, pushed my way through the
bushes and stole away across the park by the way I had come. In
the ditch inside the fence I stood for a few moments listening, but the
silence was as profound as the darkness. As quietly as I could I
climbed over the fence and dropped down into the lane. There
seemed to be not a soul moving anywhere near; nevertheless, when
I had slipped on my overcoat, instead of retracing my steps down the
lane past the entrance-gates of the cemetery, I turned to the right
and toiled up the steep hill to its termination in South Grove, where I
bore away westward and descending the long slope of West Hill,
passed the Duke of St. Albans and re-entered the Highgate Road.
It did not occur to me to look out for any conveyance. My mind
was in a whirl that seemed to communicate itself to my body and I
walked on and on like one in a dream.
The dreary miles of deserted streets were consumed unreckoned
—though still, without conscious purpose, I followed the direct road
home as a well-constructed automaton might have done. But I saw
nothing. Nor, for a time, could I be said to think coherently. My
thoughts seethed and eddied in such confusion that no product
emerged. I was conscious only of an indignant sense of shocked
decency and a loathing of Thorndyke and all his works.
Presently, however, I grew somewhat more reasonable and my
thoughts began to take more coherent shape. As a lawyer, I could
not but perceive that Thorndyke must have something definite in his
mind. He could not have done what I had seen him do without a
formal authority from the Home Secretary; and before any such
authority would have been given he would have been called upon to
show cause why the exhumation should be carried out. And such
licenses are not lightly granted. Nor, I had to admit, was Thorndyke
likely to have made the application without due consideration. He
must have had reasons for this outrageous proceeding which not
only appeared sufficient to him but which must have appeared
sufficient to the Home Secretary.
All this became by degrees clear enough to me. But yet I had not
a moment’s doubt that he had made some monstrous mistake.
Probably he had been misled by something in my diary. That
seemed to be the only possible explanation. Presently he would
discover his error—by means which I shudderingly put aside. But
when the error was discovered, the scandal would remain. It is
impossible to maintain secrecy in a case like this. In twenty-four
hours or less, all the world would know that the body of Mrs.
Monkhouse’s step-sister had been exhumed; and no subsequent
explanation would serve to destroy the effect of that announcement.
Wallingford’s dismal prophecy was about to be fulfilled.
Moreover, Thorndyke’s action amounted in effect to an open
accusation—not of Madeline or Wallingford but of Barbara, herself.
And this indignity she would suffer at my hands—at the hands of her
oldest friend! The thought was maddening. But for the outrageous
lateness of the hour, I would have gone to her at once to put her on
her guard and crave her pardon. It was the least that I could do. But
it could not be done to-night, for she would have been in bed hours
ago and her flat locked up for the night. However, I would go in the
morning at the earliest possible hour. I knew that Barbara was an
early riser and it would not be amiss if I arrived at the flat before the
maid. She must be warned at the earliest possible moment and by
me, who was the author of the mischief.
Thus, by the time that I reached my chambers I had decided
clearly what was to be done. At first, I was disposed to reject
altogether the idea of sleep. But presently, more reasonable
thoughts prevailing, I decided at least to lie down and sleep a little if I
could. But first I made a few indispensable preparations for the
morning; filled the kettle and placed it on the gas-ring, set out the
materials for a hasty breakfast, and cleaned my shoes. Then, when I
had wound the alarm clock and set it for five, I partially undressed
and crept into bed.
 Chapter XVI.
Barbara’s Message
The routine of modern life creates the habit of dividing the day
into a series of definite phases which we feel impelled to recognize
even in circumstances to which they have no real application.
Normally, the day is brought formally to an end by retirement to bed,
a process that—also normally—leads to a lapse into
unconsciousness the emergence from which marks the beginning of
another day. So, in mere obedience to the call of habit, I had gone to
bed, though, in spite of bodily fatigue, there had been no hint of any
tendency to sleep. But I might have saved myself the trouble. True,
my tired limbs stretched themselves out restfully and mere muscular
fatigue slowly wore off; but my brain continued, uselessly and
chaotically to pursue its activities only the more feverishly when the
darkness and the silence closed the avenues of impressions from
without.
Hour after hour crept by with incredible slowness, marked at each
quarter by the gentle undertone of the Treasury clock, voicing its
announcement, as it seemed, in polite protest (surely there was
never a clock that hinted so delicately and unobtrusively at the
passage of the irrevocable minutes “that perish for us and are
reckoned”). Other sound there was none to break the weary silence
of the night; but by the soft, mellow chime I was kept informed of the
birth of another day and the progress of its infancy, which crawled so
tardily in the wake of my impatience.
At last, when half-past four had struck, I threw back the bed-
clothes, and, stepping out, switched on the light and put a match to
the gas under the kettle. I had no occasion to hurry, but rather
sought to make my preparations with studied deliberation; in spite of
which I had shaved, washed and dressed and was sitting down to