![vii
Neutrophils [also known as polymorphonuclear leukocytes (PMNs) or granulocytes] are
the most abundant white cell in humans. Granulocytes and/or granulocyte precursors nor-
mally comprise ~60 % of the nucleated cells in bone marrow and blood. Mature neutrophils
have a typical circulating half-life of 6–8 h in the blood and then migrate through tissues
for ~2–3 days. Their relatively short life-span is devoted largely to surveillance for invading
microorganisms. During infection, the neutrophil life-span is extended, granulopoiesis
increases, and large numbers of neutrophils are rapidly recruited to the site(s) of infection.
Following recognition (binding) and phagocytosis of microorganisms, neutrophils utilize
an extraordinary array of oxygen-dependent and oxygen-independent microbicidal weap-
ons to destroy infectious agents. Oxygen-dependent mechanisms involve the production of
reactive oxygen species (ROS), while oxygen-independent mechanisms include degranula-
tion and release of lytic enzymes and bactericidal peptides. Inasmuch as these processes are
highly effective at killing most ingested microbes, neutrophils serve as the primary cellular
defense against infection.
The aim of Neutrophils: Methods and Protocols, Second Edition is to provide (1) a set of
protocols to assess most basic neutrophil functions, (2) protocols for investigating special-
ized areas in neutrophil research, and (3) step-by-step diagnostic assays for common neu-
trophil disorders. A wide variety of methods have been developed to assess neutrophil
function, and these methods have been instrumental in advancing our understanding of
the role of neutrophils in host defense and inflammatory disease. For those researchers and
clinicians interested in the study of neutrophils, the availability of a comprehensive source
of protocols describing the most modern methodological advances in neutrophil biology
is invaluable, as many publications do not provide information on the finer details critical
to success of a given method. As such, we have compiled a series of protocols written by
leading researchers in the field that provide detailed guidelines for establishing and per-
forming the most common neutrophil function assays. Hints of the best way to perform
these methods as well as guidance in detecting associated problems are included, so novice
investigators will also be able to effectively utilize these assays. While the volume provides
current protocols for evaluation of most basic neutrophil functions and certain specialized
functions, a section is dedicated to diagnostic assays for common neutrophil disorders.
Thus, this volume is designed for the basic researcher involved in the study of neutrophil
function and clinical investigators interested in medical aspects of neutrophil function in
health and disease.
In the second edition of Neutrophils: Methods and Protocols all of the chapters have
been updated, including many new approaches. In addition, the Second Edition contains
a number of new chapters that were not included in the First Edition. Part I is an over-
view of neutrophils and their role in host defense and inflammation. Part II describes the
most commonly used methods to isolate neutrophils from humans and other animal spe-
cies and procedures for subcellular fractionation of human neutrophils. This section also
contains a chapter that details collection and analysis of in vivo-transmigrated neutrophils.
Preface](https://image.slidesharecdn.com/80625-250419004255-68919ea7/85/Neutrophil-Methods-and-Protocols-1st-Edition-Mark-T-Quinn-13-320.jpg)
![This ebook is for the use of anyone anywhere in the United
States and most other parts of the world at no cost and with
almost no restrictions whatsoever. You may copy it, give it away
or re-use it under the terms of the Project Gutenberg License
included with this ebook or online at www.gutenberg.org. If you
are not located in the United States, you will have to check the
laws of the country where you are located before using this
eBook.
Title: The Handy Horse-book
Author: Maurice Hartland Mahon
Release date: August 21, 2020 [eBook #62993]
Most recently updated: October 18, 2024
Language: English
Credits: Produced by Julia Miller and the Online Distributed
Proofreading Team at https://www.pgdp.net (This file
was
produced from images generously made available by
The
Internet Archive/American Libraries.)
*** START OF THE PROJECT GUTENBERG EBOOK THE HANDY
HORSE-BOOK ***](https://image.slidesharecdn.com/80625-250419004255-68919ea7/85/Neutrophil-Methods-and-Protocols-1st-Edition-Mark-T-Quinn-31-320.jpg)
![PREFACE.
Finding myself a standing reference among my friends and
acquaintance on matters relating to horse-flesh, and being
constantly in the habit of giving them advice verbally and by letter, I
have been induced to comply with repeated suggestions to commit
my knowledge to paper, in the shape of a Treatise or Manual.
When I say that my experience has been practically tested on the
road, in the field, on the turf (having been formerly a steeplechase
rider, as well as now a hunting horseman), with the ribbons, and in a
cavalry regiment, I must consider that, with an ardent taste for
everything belonging to horses thus nourished for years, I must
either have sadly neglected my opportunities, or have picked up
some knowledge of the use and treatment of the animal in question.
[1]
Born and bred, I may say, in constant familiarity with a racing-
stable, and having been always devotedly attached to horses, the
wrongs of those noble animals have been prominently before my
eyes, and I have felt an anxious desire to see justice done to them,
which, I am sorry to say, according to my observation, is but too
seldom the case; indeed, I have often marvelled at the tractability of
those powerful creatures under the most perverted treatment by
their riders and drivers.
My object, therefore, in offering the following remarks, is not to
trench upon the sphere of the professional veterinary surgeon or
riding-master, but to render horse-proprietors independent of the
dictation of ignorant farriers and grooms. Intending this little work
merely as a useful manual, I have purposely avoided technicalities,
as belonging exclusively to the professional man, and endeavoured
to present my dissertations on disease in the most comprehensive
terms possible, proposing only simple remedies as far as they go;](https://image.slidesharecdn.com/80625-250419004255-68919ea7/85/Neutrophil-Methods-and-Protocols-1st-Edition-Mark-T-Quinn-37-320.jpg)
![though, for the satisfaction of my readers, I may mention that, as an
amateur, I have myself devoted much time and thought to the study
of anatomy, and that any treatment of disease herein recommended
has been carefully perused and approved by a veterinary surgeon.
Theories are excluded, and I confine myself simply to practical rules
founded on my own experience.
Hints and remarks are here offered to the general public, which, to
practical men, will appear trifling and unnecessary; but keen and
extended observation, carried on as opportunity offered, amongst all
classes and in many countries and climates, has given me an insight
into the want of reasoning exhibited by men of every station in
dealing with the noble and willing inmates of the stable, and has
assisted in suggesting the necessity for just such A B C instructions
as are herein presented by the Public’s very humble servant,
“MAGENTA.”[2]
PREFACE TO THIRD EDITION.
Increased attention having been directed to the necessity for
greater vigilance with regard to the breeding and production of good
and useful horses, many readers have expressed a wish that I would
give some decided views on these subjects; and concurring with
them as to the exigency of the case, I have ventured, in an
additional chapter in this new and Third Edition, to make a few
remarks, which, although doubtless patent to practical men, are
naturally looked for by the public in this Manual, which has been so
favourably received.
“MAGENTA.”
PREFACE TO FOURTH EDITION.
The Third Edition of this little work, published so recently as April
last, being already out of print, the Author, in presenting a new one,](https://image.slidesharecdn.com/80625-250419004255-68919ea7/85/Neutrophil-Methods-and-Protocols-1st-Edition-Mark-T-Quinn-38-320.jpg)
![PART I.
BREEDING.
A few words only of observation would I make on this subject.[3]
Palpably our horses, especially racers and hunters, are degenerating
in size and power, owing mainly, it is to be feared, to the parents
being selected more for the reputation they have gained as winners
carrying feather-weights, than for any symmetrical development or
evidence of enduring power under the weight of a man. We English
might take a useful lesson in selecting parental stock from the
French, who reject our theory of breeding from animals simply
because they have reputation in the racing calendars, and who
breed from none but those which have shape and power, as well as
blood and performance, to recommend them. They are also
particular to avoid using for stud purposes such animals as may
exhibit indications of any constitutional unsoundness.
SELECTING.
In selecting an animal, the character of the work for which he is
required should be taken into consideration. For example, in
choosing a hack, you will consider whether he is for riding or for
draught. In choosing a hunter, you must bear in mind the peculiar
nature of the country he will have to contend with.
A horse should at all times have sufficient size and power for the
weight he has to move. It is an act of cruelty to put a small horse,
be his courage and breeding ever so good, to carry a heavy man or
draw a heavy load. With regard to colour, some sportsmen say, and
with truth, that “a good horse can’t be a bad colour, no matter what
his shade.” Objection may, however, be reasonably made to pie-](https://image.slidesharecdn.com/80625-250419004255-68919ea7/85/Neutrophil-Methods-and-Protocols-1st-Edition-Mark-T-Quinn-44-320.jpg)
![balls, skew-balls, or cream-colour, as being too conspicuous,—
moreover, first-class animals of these shades are rare; nor are the
roan or mouse-coloured ones as much prized as they should be.
Bay, brown, or dark chestnuts,[4] black or grey horses, are about
the most successful competitors in the market, and may be preferred
in the order in which they are here enumerated. Very light chestnut,
bay, and white horses are said to be irritable in temper and delicate
in constitution.[5]
Mares are objected to by some as being occasionally uncertain in
temper and vigour, and at times unsafe in harness, from
constitutional irritation. More importance is attached to these
assumed drawbacks than they deserve; and though the price of the
male is generally from one-fourth to one-sixth more than that of the
female, the latter will be found to get through ordinary work quite as
well as the former.
To judge of the Age by the Teeth.—The permanent nippers, or
front teeth, in the lower jaw, are six. The two front teeth are cut and
placed at from two to three years of age; the next pair, at each side
of the middle ones, at from three and a half to four; and the corner
pair between four and a half and five years of age, when the tusks
in the male are also produced.
The marks or cavities in these nippers are effaced in the following
order:—At six years old they are worn out in the two centre teeth, at
seven in the next pair, and at eight in the corner ones, when the
horse is described as “aged.”
After this, as age advances, these nippers appear to change
gradually year by year from an oval to a more detached and
triangular form, till at twenty their appearance is completely
triangular. After six the tusks become each year more blunt, and the
grooves, which at that age are visible inside, gradually wear out.
The Hack to Ride.—A horse with a small well-shaped head seldom
proves to be a bad one; therefore such, with small fine ears, should
be sought in the first instance.](https://image.slidesharecdn.com/80625-250419004255-68919ea7/85/Neutrophil-Methods-and-Protocols-1st-Edition-Mark-T-Quinn-45-320.jpg)
![It is particularly desirable that the shoulder of a riding hack should
be light and well-placed. A high-withered horse is by no means the
best for that purpose. Let the shoulder-blades be well slanted as the
horse stands, their points light in front towards the chest. Nor should
there be too wide a front; for such width, though well enough for
draught, is not necessary in a riding-horse, provided the chest and
girth be deep.
As a matter of course the animal should be otherwise well formed,
with rather long pasterns (before but not behind),—the length of
which increases the elasticity of his movement on hard roads. His
action should be independent and high, bending the knees. If he
cannot walk well—in fact, with action so light that, as the dealers
say, “he’d hardly break an egg if he trod on it”—raising his legs
briskly off the ground, when simply led by the halter (giving him his
head)—in other words, if he walks “close to the ground”—he should
be at once rejected.
With regard to the other paces, different riders have different
fancies: the trot and walk I consider to be the only important paces
for a gentleman’s ordinary riding-horse. It is very material, in
selecting a riding-horse, to observe how he holds his head in his
various paces; and to judge of this the intending purchaser should
remark closely how he works on the bit when ridden by the rough-
rider, and he should also pay particular attention to this point when
he is himself on his back, before selection is made.[6]](https://image.slidesharecdn.com/80625-250419004255-68919ea7/85/Neutrophil-Methods-and-Protocols-1st-Edition-Mark-T-Quinn-46-320.jpg)
![The Hunter, like the hack, should be particularly well-formed
before the saddle. He should be deep in the girth, strong in the
loins, with full development of thigh, short and flat in the canon joint
from the knee to the pastern, with large flat hocks and sound fore
legs. This animal, like the road-horse, should lift his feet clear of the
ground and walk independently, with evidence of great propelling
power in the hind legs when put into a canter or gallop.
A differently-shaped animal is required for each kind of country
over which his rider has to be carried. In the midland counties and
Yorkshire, the large three-quarter or thorough-bred horse only will
be found to have pace and strength enough to keep his place. In
close countries, such as the south, south-west, and part of the north
of England, a plainer-bred and closer-set animal does best.
In countries where the fences are height jumps—a constant
succession of timber, or stone walls—one must look for a certain
angularity of hip, not so handsome in appearance, but giving greater
leverage to lift the hind legs over that description of fence.
A hunter should be all action; for if the rider finds he can be
carried safely across country, he will necessarily have more
confidence, and go straighter, not therefore requiring so much pace
to make up for round-about “gating” gaps and “craning.”[7]
BUYING.
If you propose purchasing from a dealer, take care to employ none
but a respectable man. It is also well to get yourself introduced to
such a one, by securing the good offices of some valuable customer
of his for the purpose; for such an introduction will stimulate any
dealer who values his character to endeavour by his dealings to
sustain it with his patron.](https://image.slidesharecdn.com/80625-250419004255-68919ea7/85/Neutrophil-Methods-and-Protocols-1st-Edition-Mark-T-Quinn-48-320.jpg)
![opportunity of comparison than elsewhere.
Private Purchase.—In buying from a private gentleman or
acquaintance, it is not unusual to get a horse on trial for three or
four days. Many liberal dealers, if they have faith in the animal they
want to dispose of, and in the intending purchaser, will permit the
same thing.
Warranty.—As observed under the head of “Selecting,” it is never
wise to conclude the purchase of a horse without having him
examined by a professional veterinary surgeon, and getting a
certificate of his actual state. If the animal be a high-priced one, a
warranty should be claimed from the seller as a sine qua non; and if
low-priced, a professional certificate is desirable, stating the extent
of unsoundness, for your own satisfaction.[8]
STABLING.
Ventilation is a matter of the first importance in a stable. The
means of ingress and egress of air should be always three or four
feet higher than the range of the horses’ heads, for two simple
reasons: first, when an animal comes in warm, it is not well to have
cold air passing directly on the heated surface of his body; and, in
the second place, the foul air, being the lightest, always ascends,
and you give it the readiest mode of exit by placing the ventilation
high up. The common louver window, which can never be
completely closed, is the best ordinary ventilator.
Drainage ought to be closely investigated. The drains should run
so as to remove the traps or grates outside the stable, or as far as
possible from the horses, in order to keep the effluvium away from
them. All foul litter and mass should be removed frequently during
the day; straw and litter ought not to be allowed to remain under a
horse in the daytime, unless it be considered expedient that he
should rest lying down, in which case let him be properly bedded
and kept as quiet as possible. In many cases the practice of leaving
a small quantity of litter in the stall is a fine cloak for deposit and](https://image.slidesharecdn.com/80625-250419004255-68919ea7/85/Neutrophil-Methods-and-Protocols-1st-Edition-Mark-T-Quinn-50-320.jpg)
Neutrophil Methods and Protocols 1st Edition Mark T. Quinn
- 1. Neutrophil Methods and Protocols 1st Edition Mark T. Quinn pdf download https://ebookfinal.com/download/neutrophil-methods-and- protocols-1st-edition-mark-t-quinn/ Explore and download more ebooks or textbooks at ebookfinal.com
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- 5. Neutrophil Methods and Protocols 1st Edition Mark T. Quinn Digital Instant Download Author(s): Mark T. Quinn, Frank R. DeLeo (eds.) ISBN(s): 9781627038447, 1627038442 Edition: 1 File Details: PDF, 6.95 MB Year: 2014 Language: english
- 6. Neutrophil Methods andProtocols MarkT. Quinn Frank R.DeLeo Editors SecondEdition Methods in Molecular Biology 1124
- 7. ME T H O D S I N MO L E C U L A R BI O LO G Y Series Editor John M. Walker School of Life Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK For further volumes: http://www.springer.com/series/7651
- 9. Neutrophil Methods and Protocols Second Edition Edited by MarkT. Quinn DepartmentofMicrobiologyandImmunology,MontanaStateUniversity,Bozeman,MT,USA Frank R. DeLeo Laboratory of Human Bacterial Pathogenesis, Rocky Mountain Laboratories, NationalInstituteofAllergyandInfectiousDiseases,NationalInstitutesofHealth,Hamilton,MT,USA
- 10. ISSN 1064-3745 ISSN 1940-6029 (electronic) ISBN 978-1-62703-844-7 ISBN 978-1-62703-845-4 (eBook) DOI 10.1007 /978-1-62703-845-4 Springer New York Heidelberg Dordrecht London Library of Congress Control Number: 2014930433 © Springer Science+Business Media, LLC 2014 This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. Exempted from this legal reservation are brief excerpts in connection with reviews or scholarly analysis or material supplied specifically for the purpose of being entered and executed on a computer system, for exclusive use by the purchaser of the work. Duplication of this publication or parts thereof is permitted only under the provisions of the Copyright Law of the Publisher’s location, in its current version, and permission for use must always be obtained from Springer. Permissions for use may be obtained through RightsLink at the Copyright Clearance Center. Violations are liable to prosecution under the respective Copyright Law. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. While the advice and information in this book are believed to be true and accurate at the date of publication, neither the authors nor the editors nor the publisher can accept any legal responsibility for any errors or omissions that may be made. The publisher makes no warranty, express or implied, with respect to the material contained herein. Printed on acid-free paper Humana Press is a brand of Springer Springer is part of Springer Science+Business Media (www.springer.com) Editors Mark T. Quinn Department of Microbiology and Immunology Montana State University Bozeman, MT, USA Frank R. DeLeo Laboratory of Human Bacterial Pathogenesis Rocky Mountain Laboratories National Institute of Allergy and Infectious Diseases National Institutes of Health Hamilton, MT, USA
- 11. v This volume is dedicated to Dr. Gary M. Bokoch (1954–2010) in recognition of his exten- sive contributions to neutrophil biology and GTPase signaling. Gary was a good friend and coeditor of the first edition of this volume. This volume is also dedicated to our families, who are quite patient with all of the time we spend studying neutrophils. Dedication
- 13. vii Neutrophils [also known as polymorphonuclear leukocytes (PMNs) or granulocytes] are the most abundant white cell in humans. Granulocytes and/or granulocyte precursors nor- mally comprise ~60 % of the nucleated cells in bone marrow and blood. Mature neutrophils have a typical circulating half-life of 6–8 h in the blood and then migrate through tissues for ~2–3 days. Their relatively short life-span is devoted largely to surveillance for invading microorganisms. During infection, the neutrophil life-span is extended, granulopoiesis increases, and large numbers of neutrophils are rapidly recruited to the site(s) of infection. Following recognition (binding) and phagocytosis of microorganisms, neutrophils utilize an extraordinary array of oxygen-dependent and oxygen-independent microbicidal weap- ons to destroy infectious agents. Oxygen-dependent mechanisms involve the production of reactive oxygen species (ROS), while oxygen-independent mechanisms include degranula- tion and release of lytic enzymes and bactericidal peptides. Inasmuch as these processes are highly effective at killing most ingested microbes, neutrophils serve as the primary cellular defense against infection. The aim of Neutrophils: Methods and Protocols, Second Edition is to provide (1) a set of protocols to assess most basic neutrophil functions, (2) protocols for investigating special- ized areas in neutrophil research, and (3) step-by-step diagnostic assays for common neu- trophil disorders. A wide variety of methods have been developed to assess neutrophil function, and these methods have been instrumental in advancing our understanding of the role of neutrophils in host defense and inflammatory disease. For those researchers and clinicians interested in the study of neutrophils, the availability of a comprehensive source of protocols describing the most modern methodological advances in neutrophil biology is invaluable, as many publications do not provide information on the finer details critical to success of a given method. As such, we have compiled a series of protocols written by leading researchers in the field that provide detailed guidelines for establishing and per- forming the most common neutrophil function assays. Hints of the best way to perform these methods as well as guidance in detecting associated problems are included, so novice investigators will also be able to effectively utilize these assays. While the volume provides current protocols for evaluation of most basic neutrophil functions and certain specialized functions, a section is dedicated to diagnostic assays for common neutrophil disorders. Thus, this volume is designed for the basic researcher involved in the study of neutrophil function and clinical investigators interested in medical aspects of neutrophil function in health and disease. In the second edition of Neutrophils: Methods and Protocols all of the chapters have been updated, including many new approaches. In addition, the Second Edition contains a number of new chapters that were not included in the First Edition. Part I is an over- view of neutrophils and their role in host defense and inflammation. Part II describes the most commonly used methods to isolate neutrophils from humans and other animal spe- cies and procedures for subcellular fractionation of human neutrophils. This section also contains a chapter that details collection and analysis of in vivo-transmigrated neutrophils. Preface
- 14. viii Part III encompasses protocols addressing neutrophil biochemistry, electrophysiology, signal transduction, and apoptosis. New chapters covering neutrophil microinjection and generation of mature neutrophils from induced pluripotent stem cells are now included. Part IV details methods for investigating adhesion and chemotaxis, with new chapters on evaluation of neutrophil migration through extracellular matrix and characterization of outside-in signaling via integrins. Part V provides protocols for assessing neutrophil phagocytosis and bactericidal activity, including new chapters that describe how to mea- sure phagocytosis by flow cytometry and analyze formation and function of extracellular traps. Part VI provides an extensive set of assays for evaluating NADPH oxidase priming and activation, production of reactive oxygen species, and new chapters describing analy- sis of p47phox phosphorylation and flavocytochrome b conformational changes during neutrophil activation. Part VII includes protocols to measure gene expression in neutro- phils and a new chapter on high-purity neutrophil isolation from saliva for transcriptome analysis. Finally, Part VIII provides assays for diagnosis of the most common neutrophil disorders, including an updated section on assays for myeloperoxidase and myeloperoxi- dase deficiency. In addition to the step-by-step protocols, the Notes section of each chapter provides an outstanding depot of useful and interesting information not typically published in the Methods sections of standard journal articles. We thank John M. Walker, Series Editor, and Humana Press for the opportunity to assemble an outstanding collection of articles and for help with the publication of the vol- ume. We also thank the Montana State University COBRE Center for Zoonotic and Emerging Infectious Diseases (NIH P20 GM103500) and the Intramural Research Program of the NIH, National Institutes of Allergy and Infectious Diseases, for sponsoring this volume. Finally, we thank the authors for taking time to write outstanding chapters. Bozeman, MT, USA Mark T. Quinn Hamilton, MT, USA Frank R. DeLeo Preface
- 15. ix Contents Preface.................................................................................................................... vii Contributors.................................................................................................................... xiii PART I NEUTROPHILS: AN OVERVIEW 1 The Role of Neutrophils in the Immune System: An Overview......................... 3 Harry L. Malech, Frank R. DeLeo, and Mark T. Quinn PART II NEUTROPHIL ISOLATION AND SUBCELLULAR FRACTIONATION 2 Isolation of Human Neutrophils from Venous Blood........................................ 13 William M. Nauseef 3 Neutrophil Isolation from Nonhuman Species.................................................. 19 Daniel W. Siemsen, Natalia Malachowa, Igor A. Schepetkin, Adeline R. Whitney, Liliya N. Kirpotina, Benfang Lei, Frank R. DeLeo, and Mark T. Quinn 4 Collection of In Vivo Transmigrated Neutrophils from Human Skin................. 39 Karin Christenson, Lena Björkman, Lisa Davidsson, Anna Karlsson, Per Follin, Claes Dahlgren, and Johan Bylund 5 Subcellular Fractionation of Human Neutrophils and Analysis of Subcellular Markers.................................................................. 53 Stine Novrup Clemmensen, Lene Udby, and Niels Borregaard PART III BIOCHEMISTRY, BIOLOGY AND SIGNAL TRANSDUCTION OF NEUTROPHILS 6 Rho Family and Rap GTPase Activation Assays................................................. 79 Richard T. Jennings and Ulla G. Knaus 7 Measurement of Phospholipid Metabolism in Intact Neutrophils...................... 89 Susan Sergeant and Linda C. McPhail 8 Optical Methods for the Measurement and Manipulation of Cytosolic Calcium Signals in Neutrophils ..................................................... 107 Maurice B. Hallett, Maha Al-Jumaa, and Sharon Dewitt 9 Analysis of Electrophysiological Properties and Responses of Neutrophils......... 121 Deri Morgan and Thomas E. DeCoursey 10 Assessment of Neutrophil Apoptosis................................................................. 159 David A. Dorward, Adriano G. Rossi, Ian Dransfield, and Christopher D. Lucas
- 16. x 11 Microinjection Methods for Neutrophils .......................................................... 181 Iraj Laffafian, Kimberly J. Lewis, K.Benjamin Masterman, and Maurice B. Hallett 12 Generation of Functionally Mature Neutrophils from Induced Pluripotent Stem Cells...................................................................................... 189 Colin L. Sweeney, Randall K. Merling, Uimook Choi, Debra Long Priel, Douglas B. Kuhns, Hongmei Wang, and Harry L. Malech PART IV NEUTROPHIL ADHESION AND CHEMOTAXIS 13 Neutrophil Migration Through Extracellular Matrix......................................... 209 Richard T. Jennings and Ulla G. Knaus 14 Spinning Disk Confocal Imaging of Neutrophil Migration in Zebrafish ............ 219 Pui-ying Lam, Robert S. Fischer, William D. Shin, Clare M. Waterman, and Anna Huttenlocher 15 Detection of Bidirectional Signaling During Integrin Activation and Neutrophil Adhesion................................................................................. 235 Stuart M. Altman, Neha Dixit, and Scott I. Simon PART V NEUTROPHIL PHAGOCYTOSIS AND BACTERICIDAL ACTIVITY 16 Immunofluorescence and Confocal Microscopy of Neutrophils......................... 251 Lee-Ann H. Allen 17 Expression of Genetically Encoded Fluorescent Probes to Monitor Phospholipid Dynamics in Live Neutrophils................................... 269 Benjamin E. Steinberg, Marco A.O. Magalhaes, and Sergio Grinstein 18 Quantitative Assessment of Neutrophil Phagocytosis Using Flow Cytometry..................................................................................... 279 Pontus Nordenfelt 19 Analysis of Neutrophil Bactericidal Activity....................................................... 291 Heather A. Parker, Nicholas J. Magon, Jessie N. Green, Mark B. Hampton, and Christine C. Winterbourn 20 Induction and Quantification of Neutrophil Extracellular Traps........................ 307 Alejandro Sanchez Gonzalez, Bart W. Bardoel, Christopher J. Harbort, and Arturo Zychlinsky PART VI NADPH OXIDASE AND PRODUCTION OF REACTIVE OXYGEN SPECIES 21 Measurement of Respiratory Burst Products, Released or Retained, During Activation of Professional Phagocytes ............................... 321 Johan Bylund, Halla Björnsdottir, Martina Sundqvist, Anna Karlsson, and Claes Dahlgren 22 Cell-Free NADPH Oxidase Activation Assays: “In Vitro Veritas”...................... 339 Edgar Pick Contents
- 17. xi 23 Assessment of Priming of the Human Neutrophil Respiratory Burst ................. 405 Margarita Hurtado-Nedelec, Karama Makni-Maalej, Marie-Anne Gougerot-Pocidalo, Pham My-Chan Dang, and Jamel El-Benna 24 Affinity Purification and Reconstitution of Human Phagocyte Flavocytochrome b for Detection of Conformational Dynamics in the Membrane.............................................................................................. 413 Marcia Riesselman and Algirdas J. Jesaitis 25 Evaluation of p47phox Phosphorylation in Human Neutrophils Using Phospho-Specific Antibodies.................................................................. 427 Sahra Amel Belambri, Pham My-Chan Dang, and Jamel El-Benna PART VII ANALYSIS OF NEUTROPHIL GENE EXPRESSION AND TRANSCRIPTION FACTORS 26 Genome-Scale Transcript Analyses with Human Neutrophils............................ 437 Scott D. Kobayashi, Daniel E. Sturdevant, and Frank R. DeLeo 27 Fast and Accurate Quantitative Analysis of Cytokine Gene Expression in Human Neutrophils........................................................... 451 Nicola Tamassia, Marco A. Cassatella, and Flavia Bazzoni 28 High-Purity Neutrophil Isolation from Human Peripheral Blood and Saliva for Transcriptome Analysis................................................................ 469 Flavia S. Lakschevitz and Michael Glogauer 29 Detection of Intact Transcription Factors in Human Neutrophils...................... 485 Patrick P. McDonald and Richard D. Ye PART VIII NEUTROPHIL DEFECTS AND DIAGNOSIS 30 Disorders of Neutrophil Function: An Overview............................................... 501 Mary C. Dinauer 31 Diagnostic Assays for Chronic Granulomatous Disease and Other Neutrophil Disorders....................................................................... 517 Houda Zghal Elloumi and Steven M. Holland 32 Diagnostic Assays for Myeloperoxidase and Myeloperoxidase Deficiency........... 537 William M. Nauseef Index ............................................................................................................................... 547 Contents
- 19. xiii MAHA AL-JUMAA • Neutrophil Signalling Group, School of Medicine, Cardiff University, Cardiff, UK LEE-ANN H. ALLEN • Inflammation Program and the Departments of Medicine and Microbiology, University of Iowa and the VA Medical Center, Iowa City, IA, USA STUART M. ALTMAN • Department of Biomedical Engineering, University of California, Davis, CA, USA BART W. BARDOEL • Department of Cellular Microbiology, Max Planck Institute for Infection Biology, Berlin, Germany FLAVIA BAZZONI • Section of General Pathology, Department of Pathology and Diagnostic, University of Verona, Verona, Italy SAHRA AMEL BELAMBRI • Département de Biologie, Faculté des Sciences, Université Ferhat Abbas, Sétif, Algeria LENA BJÖRKMAN • The Phagocyte Research Group, Department of Rheumatology and Inflammation Research, University of Gothenburg, Gothenburg, Sweden HALLA BJÖRNSDOTTIR • The Phagocyte Research Group, Department of Rheumatology and Inflammation Research, University of Gothenburg, Gothenburg, Sweden NIELS BORREGAARD • The Granulocyte Research Laboratory, Department of Hematology, Rigshospitalet, Copenhagen University, Copenhagen, Denmark JOHAN BYLUND • The Phagocyte Research Group, Department of Rheumatology and Inflammation Research, University of Gothenburg, Gothenburg, Sweden MARCO A. CASSATELLA • Section of General Pathology, Department of Pathology and Diagnostic, University of Verona, Verona, Italy UIMOOK CHOI • Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA KARIN CHRISTENSON • Department of Rheumatology and Inflammation Research, University of Gothenburg, Gothenburg, Sweden STINE NOVRUP CLEMMENSEN • The Granulocyte Research Laboratory, Department of Hematology, Rigshospitalet, Copenhagen University, Copenhagen, Denmark CLAES DAHLGREN • The Phagocyte Research Group, Department of Rheumatology and Inflammation Research, University of Gothenburg, Gothenburg, Sweden PHAM MY-CHAN DANG • Faculté de Medecine, Centre de Recherche Biomédicale Bichat Beaujon, INSERM U773, Paris, France LISA DAVIDSSON • The Phagocyte Research Group, Department of Rheumatology and Inflammation Research, University of Gothenburg, Gothenburg, Sweden THOMAS E. DECOURSEY • Department of Molecular Biophysics and Physiology, Rush University Medical Center, Chicago, IL, USA FRANK R. DELEO • Laboratory of Human Bacterial Pathogenesis, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, USA Contributors
- 20. xiv SHARON DEWITT • Neutrophil Signalling Group, School of Medicine, Cardiff University, Cardiff, UK MARY C. DINAUER • Departments of Pediatrics (Hematology/Oncology) and Pathology & Immunology, Washington University School of Medicine, St. Louis Children’s Hospital, St. Louis, MO, USA NEHA DIXIT • Department of Biomedical Engineering, University of California, Davis, CA, USA DAVID A. DORWARD • MRC Centre for Inflammation Research, Queen’s Medical Research Institute, University of Edinburgh, Edinburgh, UK IAN DRANSFIELD • MRC Centre for Inflammation Research, Queen’s Medical Research Institute, University of Edinburgh, Edinburgh, UK JAMEL EL-BENNA • Faculté de Medecine, Centre de Recherche Biomédicale Bichat Beaujon, INSERM U773, Paris, France HOUDA ZGHAL ELLOUMI • Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA ROBERT S. FISCHER • Cell Biology and Physiology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA PER FOLLIN • Department of Communicable Disease Control and Prevention, Västra Götaland Region, Gothenburg, Sweden MICHAEL GLOGAUER • Department of Periodontology and Matrix Dynamics Group, Faculty of Dentistry, University of Toronto, Toronto, ON, Canada ALEJANDRO SANCHEZ GONZALEZ • Department of Cellular Microbiology, Max Planck Institute for Infection Biology, Berlin, Germany MARIE-ANNE GOUGEROT-POCIDALO • Centre de Recherche Biomédicale Bichat Beaujon, INSERM U773, Paris, France JESSIE N. GREEN • Free Radical Research Group, Department of Pathology, Christchurch School of Medicine, Christchurch, New Zealand SERGIO GRINSTEIN • Cell Biology Programme, Hospital for Sick Children, Toronto, ON, Canada MAURICE B. HALLETT • Neutrophil Signalling Group, School of Medicine, Cardiff University, Cardiff, UK MARK B. HAMPTON • Free Radical Research Group, Department of Pathology, Christchurch School of Medicine, Christchurch, New Zealand CHRISTOPHER J. HARBORT • Department of Cellular Microbiology, Max Planck Institute for Infection Biology, Berlin, Germany STEVEN M. HOLLAND • Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA MARGARITA HURTADO-NEDELEC • Centre de Recherche Biomédicale Bichat Beaujon, INSERM U773, Paris, France ANNA HUTTENLOCHER • Departments of Pediatrics and Pharmacology, University of Wisconsin Medical School, Madison, WI, USA RICHARD T. JENNINGS • Conway Institute, University College Dublin, Dublin, Ireland ALGIRDAS J. JESAITIS • Department of Microbiology, Montana State University, Bozeman, MT, USA ANNA KARLSSON • The Phagocyte Research Group, Department of Rheumatology and Inflammation Research, University of Gothenburg, Gothenburg, Sweden LILIYA N. KIRPOTINA • Department of Immunology and Infectious Diseases, Montana State University, Bozeman, MT, USA Contributors
- 21. xv ULLA G. KNAUS • Conway Institute, University College Dublin, Dublin, Ireland SCOTT D. KOBAYASHI • Department of Microbiology, Molecular Biology, and Biochemistry, University of Idaho, Moscow, ID, USA DOUGLAS B. KUHNS • Neutrophil Monitoring Lab, Applied/Developmental Research Directorate, Frederick National Laboratory on Cancer Research, Frederick, MD, USA IRAJ LAFFAFIAN • Neutrophil Signalling Group, School of Medicine, Cardiff University, Cardiff, UK FLAVIA S. LAKSCHEVITZ • Matrix Dynamics Group, Faculty of Dentistry, University of Toronto, Toronto, ON, Canada PUI-YING LAM • Program in Cellular and Molecular Biology, University of Wisconsin-Madison, Madison, WI, USA BENFANG LEI • Department of Immunology and Infectious Diseases, Montana State University, Bozeman, MT, USA KIMBERLY J. LEWIS • Neutrophil Signalling Group, School of Medicine, Cardiff University, Cardiff, UK CHRISTOPHER D. LUCAS • MRC Centre for Inflammation Research, Queen’s Medical Research Institute, University of Edinburgh, Edinburgh, UK MARCO A.O. MAGALHAES • Cell Biology Programme, Hospital for Sick Children, Toronto, ON, Canada NICHOLAS J. MAGON • Free Radical Research Group, Department of Pathology, Christchurch School of Medicine, Christchurch, New Zealand KARAMA MAKNI-MAALEJ • Centre de Recherche Biomédicale Bichat Beaujon, INSERM U773, Paris, France NATALIA MALACHOWA • Laboratory of Human Bacterial Pathogenesis, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, USA HARRY L. MALECH • Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA K. BENJAMIN MASTERMAN • Neutrophil Signalling Group, School of Medicine, Cardiff University, Cardiff, UK PATRICK P. MCDONALD • Department of Pharmacology, University of Illinois, Chicago, IL, USA LINDA C. MCPHAIL • Department of Biochemistry, Wake Forest University School of Medicine, Winston-Salem, NC, USA RANDALL K. MERLING • Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA DERI MORGAN • Department of Molecular Biophysics and Physiology, Rush University Medical Center, Chicago, IL, USA WILLIAM M. NAUSEEF • Inflammation Program and Department of Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Coralville, IA, USA PONTUS NORDENFELT • Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Harvard University, Boston, MA, USA HEATHER A. PARKER • Free Radical Research Group, Department of Pathology, Christchurch School of Medicine, Christchurch, New Zealand EDGAR PICK • The Julius Friedrich Cohnheim-Minerva Center for Phagocyte Research and the Ela Kodesz Institute of Host Defense against Infectious Diseases, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel Contributors
- 22. xvi DEBRA LONG PRIEL • Neutrophil Monitoring Lab, Applied/Developmental Research Directorate, Frederick National Laboratory on Cancer Research, Frederick, MD, USA MARK T. QUINN • Department of Microbiology and Immunology, Montana State University, Bozeman, MT, USA MARCIA RIESSELMAN • Department of Microbiology, Montana State University, Bozeman, MT, USA ADRIANO G. ROSSI • MRC Centre for Inflammation Research, Queen’s Medical Research Institute, University of Edinburgh, Edinburgh, UK IGOR A. SCHEPETKIN • Department of Immunology and Infectious Diseases, Montana State University, Bozeman, MT, USA SUSAN SERGEANT • Department of Biochemistry, Wake Forest University School of Medicine, Winston-Salem, NC, USA WILLIAM D. SHIN • Cell Biology and Physiology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA DANIEL W. SIEMSEN • Department of Immunology and Infectious Diseases, Montana State University, Bozeman, MT, USA SCOTT I. SIMON • Department of Biomedical Engineering, University of California, Davis, CA, USA BENJAMIN E. STEINBERG • Cell Biology Programme, Hospital for Sick Children, Toronto, ON, Canada DANIEL E. STURDEVANT • Genomics Unit, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, USA MARTINA SUNDQVIST • The Phagocyte Research Group, Department of Rheumatology and Inflammation Research, University of Gothenburg, Gothenburg, Sweden COLIN L. SWEENEY • Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA NICOLA TAMASSIA • Section of General Pathology, Department of Pathology and Diagnostic, University of Verona, Verona, Italy LENE UDBY • The Granulocyte Research Laboratory, Department of Hematology, Rigshospitalet, Copenhagen University, Copenhagen, Denmark HONGMEI WANG • Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA CLARE M. WATERMAN • Cell Biology and Physiology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA ADELINE R. WHITNEY • Laboratory of Human Bacterial Pathogenesis, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, USA CHRISTINE C. WINTERBOURN • Free Radical Research Group, Department of Pathology, Christchurch School of Medicine, Christchurch, New Zealand RICHARD D. YE • Department of Pharmacology, University of Illinois, Chicago, IL, USA ARTURO ZYCHLINSKY • Department of Cellular Microbiology, Max Planck Institute for Infection Biology, Berlin, Germany Contributors
- 23. Part I Neutrophils: An Overview
- 25. 3 Mark T. Quinn and Frank R. DeLeo (eds.), Neutrophil Methods and Protocols, Methods in Molecular Biology, vol. 1124, DOI 10.1007/978-1-62703-845-4_1, © Springer Science+Business Media, LLC 2014 Chapter 1 The Role of Neutrophils in the Immune System: An Overview Harry L. Malech, Frank R. DeLeo, and Mark T. Quinn Abstract Neutrophils, also known as polymorphonuclear leukocytes (PMNs), have long been considered as the short-lived, nonspecific white cells that form pus—and also happen to kill invading microbes. Indeed, neutrophils were often neglected (and largely not considered) as immune cells. This historic view of neutrophils has changed considerably over the past several decades, and we know now that, in addition to playing the predominant role in the clearance of bacteria and fungi, they play a major role in shaping the host response to infection and immune system homeostasis. The change in our view of the role of neutro- phils in the immune system has been due in large part to the study of these cells in vitro. Such work has been made possible by new and/or improved methods and approaches used to investigate neutrophils. These methods are the focus of this volume. Key words Polymorphonuclear leukocyte, Granulocyte, Neutrophil methods 1 Introduction This valuable and unique book contains a compendium of methods and reviews that does much more than allow one to study the biol- ogy of neutrophils. What makes this collection of contributions so special is that it highlights and facilitates using the neutrophil as a simple, pure, single primary cell suspension model to study a remarkable array of generalized cellular functions (priming, che- motaxis and transmigration, adhesion, phagocytosis, degranula- tion, oxygen radical production, apoptosis, extracellular trap formation), biochemical pathways (GTPase activation, phospho- lipid metabolism, calcium transients, ion channel regulation, phos- phorylation events, adhesion molecule regulation), as well as specialized functions and molecules important to host defense against infection, the mediation and resolution of inflammation, and cytokine/chemokine modulation of immunity (see Fig. 1). Consideration of the array of chapter topics evokes some of the past history of inquiry into how neutrophils function and how we
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- 30. The Project Gutenberg eBook of The Handy Horse-book
- 31. This ebook is for the use of anyone anywhere in the United States and most other parts of the world at no cost and with almost no restrictions whatsoever. You may copy it, give it away or re-use it under the terms of the Project Gutenberg License included with this ebook or online at www.gutenberg.org. If you are not located in the United States, you will have to check the laws of the country where you are located before using this eBook. Title: The Handy Horse-book Author: Maurice Hartland Mahon Release date: August 21, 2020 [eBook #62993] Most recently updated: October 18, 2024 Language: English Credits: Produced by Julia Miller and the Online Distributed Proofreading Team at https://www.pgdp.net (This file was produced from images generously made available by The Internet Archive/American Libraries.) *** START OF THE PROJECT GUTENBERG EBOOK THE HANDY HORSE-BOOK ***
- 33. OPINIONS OF THE PRESS. “Most certainly the above title is no misnomer, for the ‘Handy Horse-Book’ is a manual of driving, riding, and the general care and management of horses, evidently the work of no unskilled hand.”—Bell’s Life. “As cavalry officer, hunting horseman, coach proprietor, whip, and steeplechase rider, the author has had long and various experience in the management of horses, and he now gives us the cream of his information in a little volume, which will be to horse-keepers and horse-buyers all that the ‘Handy Book on Property Law,’ by Lord St Leonards, has for years past been to men of business. It does not profess to teach the horse-keeper everything that concerns the beast that is one of the most delicate as well as the noblest of animals; but it supplies him with a number of valuable facts, and puts him in possession of leading principles.”— Athenæum. “The writer shows a thorough knowledge of his subject, and he fully carries out the object for which he professes to have undertaken his task—namely, to render horse-proprietors independent of the dictations of ignorant farriers and grooms.”— Observer. “We need only say that the work is essentially a multum in parvo, and that a book more practically useful, or that was more required, could not have possibly been written.”—Irish Times. “He propounds no theories, but embodies in simple and untechnical language what he has learned practically; and a perusal of the volume will at once testify that he is fully qualified for the task; and so skilfully is the matter condensed that there is scarcely a single sentence which does not convey sound and valuable information.”—Sporting Gazette. “We can cordially recommend it as a book especially suited to the general public, and not beneath the attention of ‘practical men.’”—The Globe. “Contains a very great modicum of information in an exceedingly small space.... There can be little doubt that it will, when generally known, become the established vade mecum of the fox-hunter, the country squire, and the trainer.”— Army and Navy Gazette. “A useful little work.... In the first part he gives just the amount of information that will enable a man to work his horse comfortably, check his groom, and generally know what he is about when riding, driving, or choosing gear.”— Spectator. “This is a book to be read and re-read by all who take an interest in the noble animal, as it contains a most comprehensive view of everything appertaining to
- 34. horse-flesh; and is, moreover, as fit for the library and drawing-room as it is for the mess-table or the harness-room.”—Sporting Magazine. “By all means buy the book; it will repay the outlay.”—Land and Water. DRAWING COVER THE HANDY HORSE-BOOK
- 35. OR PRACTICAL INSTRUCTIONS IN DRIVING, RIDING, AND THE GENERAL CARE AND MANAGEMENT OF HORSES BY A CAVALRY OFFICER FOURTH EDITION, REVISED AND ENLARGED With Engravings WILLIAM BLACKWOOD AND SONS EDINBURGH AND LONDON MDCCCLXVIII The Right of Translation is reserved TO MAJOR-GENERAL LORD GEORGE PAGET, C.B. Inspector-General of Cavalry,
- 36. SON OF THE DISTINGUISHED HORSEMAN AND HERO WHO COMMANDED THE CAVALRY AT WATERLOO, AND HIMSELF A LEADER AMONG THE “IMMORTAL SIX HUNDRED,” THIS BOOK IS BY PERMISSION INSCRIBED, IN TRIBUTE TO HIS SOLDIERLY QUALITIES, AND TO HIS CONSIDERATION FOR THE NOBLE ANIMAL WHICH HAS CARRIED THE BRITISH CAVALRY THROUGH SO MANY DANGERS TO SO MANY TRIUMPHS, BY HIS LORDSHIP’S OBEDIENT SERVANT, “MAGENTA.”
- 37. PREFACE. Finding myself a standing reference among my friends and acquaintance on matters relating to horse-flesh, and being constantly in the habit of giving them advice verbally and by letter, I have been induced to comply with repeated suggestions to commit my knowledge to paper, in the shape of a Treatise or Manual. When I say that my experience has been practically tested on the road, in the field, on the turf (having been formerly a steeplechase rider, as well as now a hunting horseman), with the ribbons, and in a cavalry regiment, I must consider that, with an ardent taste for everything belonging to horses thus nourished for years, I must either have sadly neglected my opportunities, or have picked up some knowledge of the use and treatment of the animal in question. [1] Born and bred, I may say, in constant familiarity with a racing- stable, and having been always devotedly attached to horses, the wrongs of those noble animals have been prominently before my eyes, and I have felt an anxious desire to see justice done to them, which, I am sorry to say, according to my observation, is but too seldom the case; indeed, I have often marvelled at the tractability of those powerful creatures under the most perverted treatment by their riders and drivers. My object, therefore, in offering the following remarks, is not to trench upon the sphere of the professional veterinary surgeon or riding-master, but to render horse-proprietors independent of the dictation of ignorant farriers and grooms. Intending this little work merely as a useful manual, I have purposely avoided technicalities, as belonging exclusively to the professional man, and endeavoured to present my dissertations on disease in the most comprehensive terms possible, proposing only simple remedies as far as they go;
- 38. though, for the satisfaction of my readers, I may mention that, as an amateur, I have myself devoted much time and thought to the study of anatomy, and that any treatment of disease herein recommended has been carefully perused and approved by a veterinary surgeon. Theories are excluded, and I confine myself simply to practical rules founded on my own experience. Hints and remarks are here offered to the general public, which, to practical men, will appear trifling and unnecessary; but keen and extended observation, carried on as opportunity offered, amongst all classes and in many countries and climates, has given me an insight into the want of reasoning exhibited by men of every station in dealing with the noble and willing inmates of the stable, and has assisted in suggesting the necessity for just such A B C instructions as are herein presented by the Public’s very humble servant, “MAGENTA.”[2] PREFACE TO THIRD EDITION. Increased attention having been directed to the necessity for greater vigilance with regard to the breeding and production of good and useful horses, many readers have expressed a wish that I would give some decided views on these subjects; and concurring with them as to the exigency of the case, I have ventured, in an additional chapter in this new and Third Edition, to make a few remarks, which, although doubtless patent to practical men, are naturally looked for by the public in this Manual, which has been so favourably received. “MAGENTA.” PREFACE TO FOURTH EDITION. The Third Edition of this little work, published so recently as April last, being already out of print, the Author, in presenting a new one,
- 39. feels called upon gratefully to acknowledge this unusual mark of favour on the part of the public. London, November 1867.
- 40. CONTENTS. PAGE PART I. BREEDING, 1 SELECTING, 2 BUYING, 6 STABLING, 8 GROOMING, 12 HALTERING, 16 CLOTHING, 18 FEEDING, 20 WATERING, 25 GRAZING, 26 TRAINING, 28 EXERCISING, 31 WORK, 33 BRIDLING, 38 SADDLING, 43 RIDING, 49 HARNESSING, 56 DRIVING, 65 DRAWING, 72 SHOEING, 75 VICE, 84 SELLING, 89 CAPRICE, 90 IRISH HUNTERS, AND THE BREEDING OF GOOD HORSES, 93 PART II.
- 41. DISEASES, 101 OPERATIONS, 102 TO GIVE A BALL, 104 TO GIVE A DRENCH, 105 PURGING, 106 THE PULSE, 109 DISEASES OF THE HEAD AND RESPIRATORY ORGANS, 109 DISEASES OF THE DIGESTIVE AND URINARY ORGANS, 120 DISEASES OF THE FEET AND LEGS, 127 LOTIONS, PURGES, BLISTERS, ETC., 158 INDEX, 164
- 42. LIST OF PLATES. DRAWING COVER, frontispiece. THE HACK, page 4 THE WEIGHT-CARRYING HUNTER, ” 6 RIDING AT IT, ” 53 THE PROPER FORM, ” 95 PREPARATORY CANTER, ” 99
- 44. PART I. BREEDING. A few words only of observation would I make on this subject.[3] Palpably our horses, especially racers and hunters, are degenerating in size and power, owing mainly, it is to be feared, to the parents being selected more for the reputation they have gained as winners carrying feather-weights, than for any symmetrical development or evidence of enduring power under the weight of a man. We English might take a useful lesson in selecting parental stock from the French, who reject our theory of breeding from animals simply because they have reputation in the racing calendars, and who breed from none but those which have shape and power, as well as blood and performance, to recommend them. They are also particular to avoid using for stud purposes such animals as may exhibit indications of any constitutional unsoundness. SELECTING. In selecting an animal, the character of the work for which he is required should be taken into consideration. For example, in choosing a hack, you will consider whether he is for riding or for draught. In choosing a hunter, you must bear in mind the peculiar nature of the country he will have to contend with. A horse should at all times have sufficient size and power for the weight he has to move. It is an act of cruelty to put a small horse, be his courage and breeding ever so good, to carry a heavy man or draw a heavy load. With regard to colour, some sportsmen say, and with truth, that “a good horse can’t be a bad colour, no matter what his shade.” Objection may, however, be reasonably made to pie-
- 45. balls, skew-balls, or cream-colour, as being too conspicuous,— moreover, first-class animals of these shades are rare; nor are the roan or mouse-coloured ones as much prized as they should be. Bay, brown, or dark chestnuts,[4] black or grey horses, are about the most successful competitors in the market, and may be preferred in the order in which they are here enumerated. Very light chestnut, bay, and white horses are said to be irritable in temper and delicate in constitution.[5] Mares are objected to by some as being occasionally uncertain in temper and vigour, and at times unsafe in harness, from constitutional irritation. More importance is attached to these assumed drawbacks than they deserve; and though the price of the male is generally from one-fourth to one-sixth more than that of the female, the latter will be found to get through ordinary work quite as well as the former. To judge of the Age by the Teeth.—The permanent nippers, or front teeth, in the lower jaw, are six. The two front teeth are cut and placed at from two to three years of age; the next pair, at each side of the middle ones, at from three and a half to four; and the corner pair between four and a half and five years of age, when the tusks in the male are also produced. The marks or cavities in these nippers are effaced in the following order:—At six years old they are worn out in the two centre teeth, at seven in the next pair, and at eight in the corner ones, when the horse is described as “aged.” After this, as age advances, these nippers appear to change gradually year by year from an oval to a more detached and triangular form, till at twenty their appearance is completely triangular. After six the tusks become each year more blunt, and the grooves, which at that age are visible inside, gradually wear out. The Hack to Ride.—A horse with a small well-shaped head seldom proves to be a bad one; therefore such, with small fine ears, should be sought in the first instance.
- 46. It is particularly desirable that the shoulder of a riding hack should be light and well-placed. A high-withered horse is by no means the best for that purpose. Let the shoulder-blades be well slanted as the horse stands, their points light in front towards the chest. Nor should there be too wide a front; for such width, though well enough for draught, is not necessary in a riding-horse, provided the chest and girth be deep. As a matter of course the animal should be otherwise well formed, with rather long pasterns (before but not behind),—the length of which increases the elasticity of his movement on hard roads. His action should be independent and high, bending the knees. If he cannot walk well—in fact, with action so light that, as the dealers say, “he’d hardly break an egg if he trod on it”—raising his legs briskly off the ground, when simply led by the halter (giving him his head)—in other words, if he walks “close to the ground”—he should be at once rejected. With regard to the other paces, different riders have different fancies: the trot and walk I consider to be the only important paces for a gentleman’s ordinary riding-horse. It is very material, in selecting a riding-horse, to observe how he holds his head in his various paces; and to judge of this the intending purchaser should remark closely how he works on the bit when ridden by the rough- rider, and he should also pay particular attention to this point when he is himself on his back, before selection is made.[6]
- 47. THE HACK Respecting soundness, though feeling fully competent myself to judge of the matter, I consider the half-guinea fee to a veterinary surgeon well-laid-out money, to obtain his professional opinion and a certificate of the state of an animal, when purchasing a horse of any value. The Hack for Draught ought to be as well formed as the one just described; but a much heavier shoulder and forehand altogether are admissible. No one should ever for a moment think of putting any harness- horse into a private vehicle, no matter what his seller’s recommendation, without first having him out in a single or double break, as the case may be, and seeing him driven, as well as driving him himself, to make acquaintance with the animal—in fact, to find him out.
- 48. The Hunter, like the hack, should be particularly well-formed before the saddle. He should be deep in the girth, strong in the loins, with full development of thigh, short and flat in the canon joint from the knee to the pastern, with large flat hocks and sound fore legs. This animal, like the road-horse, should lift his feet clear of the ground and walk independently, with evidence of great propelling power in the hind legs when put into a canter or gallop. A differently-shaped animal is required for each kind of country over which his rider has to be carried. In the midland counties and Yorkshire, the large three-quarter or thorough-bred horse only will be found to have pace and strength enough to keep his place. In close countries, such as the south, south-west, and part of the north of England, a plainer-bred and closer-set animal does best. In countries where the fences are height jumps—a constant succession of timber, or stone walls—one must look for a certain angularity of hip, not so handsome in appearance, but giving greater leverage to lift the hind legs over that description of fence. A hunter should be all action; for if the rider finds he can be carried safely across country, he will necessarily have more confidence, and go straighter, not therefore requiring so much pace to make up for round-about “gating” gaps and “craning.”[7] BUYING. If you propose purchasing from a dealer, take care to employ none but a respectable man. It is also well to get yourself introduced to such a one, by securing the good offices of some valuable customer of his for the purpose; for such an introduction will stimulate any dealer who values his character to endeavour by his dealings to sustain it with his patron.
- 49. THE WEIGHT-CARRYING HUNTER Auction.—An auction is a dangerous place for the uninitiated to purchase at. If, however, it should suit you to buy in that manner, the best course to pursue is to visit the stables on the days previous to the sale, for in all well-regulated repositories the horses are in for private inspection from two to three days before the auction-day. Taking, if possible, one good judge with you, eschewing the opinions of all grooms and others—in fact, fastening the responsibility of selection on the one individual—make for yourself all the examination you possibly can, in or out of stable, of the animal you think likely to suit you. There is generally a way of finding out some of the antecedents of the horses from the men about the establishment. Fairs.—To my mind it is preferable to purchase at fairs rather than at an auction: indeed, a judge will there have much more
- 50. opportunity of comparison than elsewhere. Private Purchase.—In buying from a private gentleman or acquaintance, it is not unusual to get a horse on trial for three or four days. Many liberal dealers, if they have faith in the animal they want to dispose of, and in the intending purchaser, will permit the same thing. Warranty.—As observed under the head of “Selecting,” it is never wise to conclude the purchase of a horse without having him examined by a professional veterinary surgeon, and getting a certificate of his actual state. If the animal be a high-priced one, a warranty should be claimed from the seller as a sine qua non; and if low-priced, a professional certificate is desirable, stating the extent of unsoundness, for your own satisfaction.[8] STABLING. Ventilation is a matter of the first importance in a stable. The means of ingress and egress of air should be always three or four feet higher than the range of the horses’ heads, for two simple reasons: first, when an animal comes in warm, it is not well to have cold air passing directly on the heated surface of his body; and, in the second place, the foul air, being the lightest, always ascends, and you give it the readiest mode of exit by placing the ventilation high up. The common louver window, which can never be completely closed, is the best ordinary ventilator. Drainage ought to be closely investigated. The drains should run so as to remove the traps or grates outside the stable, or as far as possible from the horses, in order to keep the effluvium away from them. All foul litter and mass should be removed frequently during the day; straw and litter ought not to be allowed to remain under a horse in the daytime, unless it be considered expedient that he should rest lying down, in which case let him be properly bedded and kept as quiet as possible. In many cases the practice of leaving a small quantity of litter in the stall is a fine cloak for deposit and
- 51. urine left unswept underneath, emitting that noxious ammonia with which the air of most stables is so disagreeably impregnated that on entering them from the fresh air you are almost stifled. Masters who object to their horses standing on the bare pavement can order that, after the stall is thoroughly cleaned and swept out, a thin layer of straw shall be laid over the stones during the daytime. In dealers’ and livery stables, and indeed in some gentlemen’s, the pavement is sanded over, which has a nice appearance, and prevents slipping. When the foul litter is abstracted, and the straw bedding taken from under the horse, none of it should be pushed away under the manger; let it be entirely removed: and in fair weather, or where a shed is available, the bedding should be shaken out, to thoroughly dry and let the air pass through it. Wheaten is more durable than oaten straw for litter: but the fibre of the former is so strong that it will leave marks on the coat of a fine-skinned animal wherever it may be unprotected by the clothing; however, this is not material. Light should be freely admitted into stables, not only that the grooms may be able to see to clean the horses properly, and to do all the stable-work, but if horses are kept in the dark it is natural that they should be more easily startled when they go into full daylight,—and such is always the consequence of badly-lighted stables. Of course, if a horse is ailing, and sleep is absolutely necessary for him, he should be placed separate in a dark quiet place. Stalls should be wide, from six to seven feet across if possible, yielding this in addition to other advantages, that if the partitions are extended by means of bars to the back wall, either end stall can be turned into a loose-box sufficiently large to serve in an emergency. A Loose-Box is unquestionably preferable to a stall (in which a horse is tied up all the time he is not at work in nearly the same position), and is indispensable in cases of illness. Loose-boxes
- 52. should be paved with narrow bricks; and when prepared for the reception of an animal whose shoes have been removed, the floor should be covered with sawdust or tan, or either of these mixed with fine sandy earth, or, best of all, peat-mould when procurable,—any of which, where the indisposition is confined to the feet only, may be kept slightly moistened with water to cool them. In cases of general illness, straw should be used for bedding; and where the poor beast is likely to injure himself in paroxysms of pain, the walls or partitions should be well padded in all parts within his reach, and as a further precaution let the door be made to open outwards, and be fastened by a bolt, as latches sometimes cause accidents. Partitions should be carried high enough towards the head to prevent the horses from being able to bite one another, or get at each other’s food. With regard to stable-kickers, see the remarks on this subject under the head of “Vice” (page 85). Racks and Mangers are now made of iron, so that horses can no longer gnaw away the manger piecemeal. Another improvement is that of placing the rack on a level with and beside the manger, instead of above the horses’ heads; but notwithstanding this more reasonable method of feeding hay when whole, it is far preferable to give it as manger-food cut into chaff. Flooring.—In the construction of most stables a cruel practice is thoughtlessly adopted by the way of facilitating drainage (and in dealers’ stables to make horses look large), viz., that of raising the paving towards the manger considerably above the level of the rear part. It should be borne in mind that the horse is peculiarly sensitive to any strain on the insertions of the back or flexor tendons of his legs. Thus in stalls formed as described, you will see the creature endeavouring to relieve himself by getting his toes down between the flags or stones (if the pavement will admit) with the heels resting upon the edges of them; and if the fastening to the head be
- 53. long enough he will draw back still farther, until he can get his toes down into the drain-channel behind his stall, with the heels upon the opposite elevation of the drain. Proper pavement in your stable will help to alleviate a tendency towards what is called “clap of the back sinew.”—See page 143. The slope of an inch and a half or two inches is sufficient for purposes of drainage in paving stables; but if the drainage can be managed so as to allow of the flooring being made quite level, so much the better. Should my reader be disposed to build stabling, he cannot do better than consult the very useful and practical work entitled ‘Stonehenge, or the Horse in the Stable and in the Field.’ The horse being a gregarious animal, and much happier in society than alone, will, in the absence of company of his own species, make friends with the most sociable living neighbour he can find. A horse should not be left solitary if it can be avoided. Dogs should never be kept in the stable with horses, or be permitted to be their playfellows, on account of the noxious emissions from their excrement. Cats are better and more wholesome companions. GROOMING. I do not profess to teach grooms their business, but to put masters on their guard against the common errors and malpractices of that class; and with a view to that end, two or three general rules are added which a master would do well to enforce on a groom when hiring him, as binding, under pain of dismissal. 1. Never to doctor a horse himself, but to acquaint his master immediately with any accident, wound, or symptom of indisposition about the animal, that may come under his observation, and which, if in existence, ought not to fail to attract the attention of a careful,
- 54. intelligent servant during constant handling of and attendance on his charge. 2. Always to exercise the horses in the place appointed by his master for the purpose, and never to canter or gallop them. 3. To stand by while a horse is having its shoes changed or removed, and see that any directions he may have received on the subject are carried out. 4. Never to clean a horse out of doors. These rules are recommended under a just appreciation of that golden one, “Prevention is better than cure.” If the master is satisfied with an ill-groomed horse, nine-tenths of the grooms will be so likewise; therefore he may to a great extent blame himself if his bearer’s dressing is neglected. Grooms are especially fond of using water in cleaning the horse (though often rather careful how they use it with themselves, either inside or out): it saves them trouble, to the great injury of the animal. The same predominating laziness which prompts them to use water for the removal of mud, &c., in preference to employing a dry wisp or brush for the purpose, forbids their exerting themselves to employ the proper means of drying the parts cleaned by wet. They will have recourse to any expedient to dry the skin rather than the legitimate one of friction. Over the body they will place cloths to soak up the wet; on the legs they will roll their favourite bandages. It is best, therefore, to forbid the use of water above the hoof for the purpose of cleaning—except with the mane and tail, which should be properly washed with soap and water occasionally. When some severe work has been done, so as to occasion perspiration, the ears should not be more neglected than the rest of the body; and when they are dried by hand-rubbing and pulling, the horse will feel refreshed. As already recommended, cleaning out of doors should be forbidden. If one could rely on the discretion of servants, cleaning
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