The articles published in this volume are based on the papers del ivered at a conference on the Role of Metal Ions in Biological Systems held November 20 and 21, 1972, at Argonne National Laboratory. The purpose of the conference was to present to an interdiscipl inary audience of physical scientists some recent developments illustrating the chemical and environ mental participation of the heavy metal ions in the biological system. The invited speakers at the conference are special ists in the fields they describe, and the articles presented here are at a level of interest to readers with backgrounds in physical sciences who are not necessari Iy doing research in the areas described. The articles are referenced through 1972, and in some cases early 1973, and thus should also be of value to research workers. It is hoped that the book wi II be of particular interest to chemists, biologists, workers in the fields of en vironmental science and publ ic health, as wei I as graduate and senior undergraduate students in these discipl ines. The conference was sponsored by the Central States Universities, Inc., a consortium of sixteen midwestern univer sities, the Center for Educational Affairs, Argonne National Laboratory, and the United States Atomic Energy Commission. It is my pleasure to thank the members of the conference committee for their ideas and active help in organizing the conference."

With the rapid spread of drug use and abuse in this country over the last decade, events relating to drugs have become increasingly prominent in the court of law as well as in society in general. It is anticipated that throughout the 1980s this problem will continue to be one of the major social problems in the United States. With the complexity of the Federal and state laws and their interrelationships, and with the increasing demand for the expert chemist or toxicologist, or both, in the courtroom, more education for these people relating to drugs must be implemented. The prosecuting and defense attorneys are also in need of accurate toxicological information to adequately handle cases in which drugs playa role. This book is intended to be a practical as well as an educational book-a resource tool for the active forensic chemist, as well as for the student. It is with the forensic chemist in mind that the material on controlled substances and substances found in cases of drug abuse has been arranged in a systematic order for quick and easy reference. This section includes synonyms, pharma ceutical preparations, biochemistry and/or pharmacognosy, toxicology and/or pharmacology, and how the substance is controlled under Federal law. Under the concept of scientific consideration as compared with the Federal law, this book enables one to deal with some of the major conflicts in the American legal system concerning scientific evidence and how it is presented in a court of law."

Tiselius demonstrated that the immunologically active components of immune sera migrated electrophoretically in the gamma globulin region. His findings illuminated the classic observations of Jenner regarding development of resistance to infection, and those of von Pirquet, Pasteur, and Arthus regarding the transfer and specificity of resistance. Conceptual integration of these observations provided the impetus for the present modern era of immunology. Subsequent to Tiselius's work, multiple, rapid advances have occurred in the study of congenital and acquired immune deficiency states in mice, chickens, and humans. These studies have readily demonstrated that the immunologic ability of an organ ism to protect itself from environmental influences is a prerequisite for survival. Indeed, this necessity for protection from microenvironmental influences has promoted the evolu tionary development of immunologic diversification, namely, host dependence upon a sophisticated, multifaceted network of cells and effector mechanisms responsible for the clearance and neutralization of toxins and potentially harmful pathogens. The obligate dependence of animals upon the functional integrity of their immunologic systems is illus trated by the ready invasion of ubiquitous organisms when the host is in a state of immune defense derangement. Nevertheless, derangements in immune function can range from par tial to complete and can be compatible with survival. The consequences of such derange ments run the gamut from subclinical disease to inevitable mortality."

Elastic filaments refer mainly to titin, the largest of all known proteins. Titin was discovered initially in muscle cells, where it interconnects the thick filament with the Z-line. Titin forms a molecular spring that is responsible for maintaining the structural integrity of contracting muscle, ensuring efficient muscle contraction. More recently, it has become clear that titin is not restricted to muscle cells alone. For example, titin is found in chromosomes of neurons and also in blood platelets. This topic is fast becoming a focal point for research in understanding viscoelastic properties at the molecular, cellular, and tissue levels. In titin may lie a generic basis for biological viscoelasticity. It has become clear that titin may hold the key to certain clinical anomalies. For example, it is clear that titin-based ventricular stiffness is modulated by calcium and that titin is responsible for the altered stiffness in cardiomyopathies. It is also clear from evidence from a group of Finnish families that titin mutations may underlie some muscular dystrophies and that with other mutations chromatids fail to separate during mitosis. Thus, it is clear that this protein will have important clinical implications stemming from its biomechanical role. One aspect of this field is the bringing together of bioengineers with clinical researchers and biologists. Genetic and biochemical aspects of titin-related proteins are being studied together with front-line engineering approaches designed to measure the mechanics of titin either in small aggregates or in single molecules.

A variety of metabolic processes are known to be intimately involved in the maintenance of cellular structure and function. It has also become clear that metabolic events involved in the synthesis and hydrolysis of ATP as well as for the synthesis of proteins and phospholipids are essential for cellular health. The regulation of cell function is generally achieved through participation of a wide variety of hormones and different signal transduction mechanisms for the activation/deactivation of some specific metabolic processes. In this regard cyclic AMP and calcium seem to play a crucial role. Various hormones are also known to affect the genetic machinery of all the cell; however, the exact signals for genetic control of cellular function are not well defined. In particular, the sequence of events concerned with remodelling of different types of cells under various pathological situations is poorly understood. In this book we have therefore dealt with some of these issues from biochemical, molecular biological, physiological, and pharmacological viewpoints. Special emphasis has been laid on understanding heart function and metabolism in health and disease in general, and cardiac hypertrophy, heart failure, and ischemic heart disease in particular. It is hoped that this multidisciplinary information will be of value to basic scientists and clinical investigators.

The field of genetic toxicology is a relatively new one which grew out of the studies of chemical mutagenesis and modern toxicology. Considering that systematic practices to detect chemical mutagenesis are only a little over thirty years old, this field has evolved very rapidly with an abundance of methods for identifying chemical mutagens. To evalulate the usefulness of the methods and to select the assay which will yield the most important information under practical conditions requires the broad experience such as that which Dr. Brusick has acquired over the last decade. Since this field is expanding very rapidly and new test ing methods are being recognized, it should be kept in mind that revisions may have to be made during the next five to ten years. The need for such a book has been obvious to us, particularly since training courses and workshops on genetic toxicology are being organized which find it beneficial to utilize established guidelines and since the reports in the literature do not always describe in detail how the work was carried out during laboratory testing. In addition to his broad background in genetics, Dr. Brusick has had much practical experience, having organized and directed the most extensive laboratory for mutagen testing. I am most pleased to see this volume, for which there is an ever-increasing need. Alexander Hollaender Associated Universities, Inc. 1717 Massachusetts Avenue, N.W."

The best protection against environmental mutagens is to identify them before they ever come into general use. But it is always possible that some substance will escape detection and affect a large number of persons without this being realized until later generations. This article considers ways in which such a genetic emergency might be promptly detected. A mutation-detecting system should be relevant in that it tests for effects that are as closely related as possible to those that are feared. It should be sensitive enough to detect a moderate increase in mutation rate, able to discover the increase promptly before more damage is done, responsive to various kinds of mutational events, and designed in such a way as to maxi- mize the probability that the Gause of an increase can be found. Methods based on germinal mutation necessarily involve enormous numbers of persons and tests. On the other hand, with somatic mutations the individual cell becomes the unit of measurement rather than the in- dividual person. For this reason, I think that somatic tests are preferable to germinal tests, despite the fact that it is germinal mutations which are feared.

The traditional approach to diagnosis of virus infectians by isolation of the causative virus is usually both slow and expensive. More recently, the emphasis has been on the direct detection of viruses or viral antigens in clinical specimens. This can be done using established techniques such as immunofluorescence or electron microscopy, or by newly developed biochemical methods. The purpose of this meeting was to review these and other developments in the laboratory diagnosis of virus infections. We would like to thank all those who contributed to the success of this meeting. In particular, we are grateful to the CEC for financial sponsorship, to Professor C. Dow, Director of the Veterinary Research Laboratories, for help in organising the meeting, and to Miss B. Hamilton for her excellent typing of the manuscripts. M S McNulty J B McFerran VII CONTENTS Page Preface V VIRAL DIAGNOSIS BY TIMMUNOFLUORESCENCE P. S. Gardner APPLICATIONS OF IMMUNOFLUORESCENCE IN VETERINARY VIRAL DIAGNOSIS M. S. McNulty and G. M. Allan 15 TAGGING OF VIRAL ANTIGENS USING IMMUNOPEROXIDASE . AND IMMUNOOOLD TEGINlQUES R. Ducatelle, F. Castryck and J. Hoorens 27 DETECTION OF BVD VIRUS IN VlRAEMIC CATTLE BY AN INDIRECT IMMUNOPEROXIDASE TEGINIQUE A. Meyling 37 VIRAL DIAGNOSIS BY ELECTRON MICROSCOPY J. D. Almeida 47 CAPTURE ASSAYS FOR THE DETECTION OF VIRUS-SPECIFIC IgM ANTIBODY R. S. Tedder 60 DETECTION . AND IDENTIFICATION OF PAPILLOMAVIRUSES IN BENIGN . AND MALIGNANT 'TIlmJRS OF CATTLE M. S. Campo 72 DETECTION OF VIRAL NUCLEIC ACIDS IN CELL CULTURES .

This publication contains the proceedings of a Seminar "Beef production from different dairy breeds and dairy beef crosses," held in Ireland on April 13-15, 1981, under the auspices of the Commission of the European Communities (CEC) as part of the European Communities (EC) programme for beef production research. The CEC wishes to thank those representatives of Ireland who took responsibility for the organisation and conduct of this Seminar, notably Professor E.P. Cunningham, Dr. G.J. More O'Ferrall (local organiser), Dr. Patricia McGlaughlin and In particular, thanks are due to Dr. McGlaughlin Mr. R. Barlow. and Mr. Barlow for their recording of the discussions. Professor Ian Gordon of the Faculty of Agriculture, kindly made available the facilities of Lyons House, University College, for the Seminar. Thanks are also accorded to the Chairmen of the Sessions Professor D. Smidt, Dr. R.B. Thiessen, Professor A. Neimann Sorensen, Professor E.P. Cunningham, and to all the participants who presented papers and took part in the discussions. X OBJECTIVES The aims of the Seminar were to review recent comparisons of Holstein and Friesian strains with other dairy breeds for beef and veal production; to look at the use of beef breeds for crossing on dairy herds in various EEC countries, and to examine the economic and genetic balance between milk and beef traits in dual purpose bull testing and selection."

Where do you begin to look for a recent, authoritative article on the diagnosis or management of a particular malignancy? The few general oncology textbooks are generally out of date. Single papers in specialized journals are informative but seldom comprehensive; these are more often preliminary reports on a very limited number of patients. Certain general journals frequently publish good indepth reviews of cancer topics, and published symposium lectures are often the best overviews available. Unfor tunately, these reviews and supplements appear sporadically, and the reader can never be sure when a topic of special interest will be covered. Cancer Treatment and Research is a series of authoritative volumes which aim to meet this need. It is an attempt to establish a critical mass of oncology literature covering virtually all oncology topics, revised frequently to keep the coverage up to date, easily available on a single library shelf or by a single personal subscription. We have approached the problem in the following fashion. First, by dividing the oncology literature into specific subdivisions such as lung can cer, genitourinary cancer, pediatric oncology, etc. Second, by asking emi nent authorities in each of these areas to edit a volume on the specific topic on an annual or biannual basis. Each topic and tumor type is covered in a volume appearing frequently and predictably, discussing current diagnosis, staging, markers, all forms of treatment modalities, basic biology, and more."

The twentieth century has seen a remarkable evolution of environmental health and environmental protection concerns and concepts in the United States. As a teacher of Environmental Health since the late 1950s, I have witnessed the many twists and turns that have marked the latter half of the century, and have had to seek continuously to explain these phenomena to my students in some rational manner. We have witnessed the following and more: great progress in controlling acute infectious diseases through successes in drinking water treatment and food processing; the emergence of greater concern with trace chemicals in air and water and their role in chronic disease causation; conflicting attitudes toward miraculous chemicals such as DDT (which promised relief from arthropod-borne scourges, then came to be seen as another chemical threat to our children and our environment); then the reemerging concern with infectious diseases precipitated by blood-borne pathogens such as HIV. All this occurred against a backdrop of scientific uncertainty and amid failures of risk assessment and risk communication, together with press sensationalism-from "mad cow disease" to "flesh eating" streptococci. No wonder the public is confused.

Take a disease of complex pathology with inflammatory and neoplastic features, which affects lymphoid and neural tissues, belonging to a disease group which killed one chicken in five, and which defied efforts to understand and control it for lOre than 50 years, and one can begin to appreciate the interest Marek's disease has received. Canpound these characteristics with the finding of the causal herpesvirus, its recog nition as the neoplasm first discovered to be so caused, and its pre vention by vaccination, and the special place of Marek's disease in veterinary medicine and comparative oncology becomes clear. This book sets out to provide an authoritative and corrprehensive account of knowledge of Marek's disease and its control. I hope that it will be of value to veterinary research workers, teachers and students who need information about the disease, to veterinarians, poultrymen and vaccine manufacturers who have to diagnose and control it, and to oncologists in other fields interested in comparative aspects. other reviews of the disease exist, of course, but this is the first multi authored book devoted to the subject."

The "Groupe International de Recherche sur l'Infinitesimal" (GIRI) was created in 1986 by Professor Madeleine Bastide and Doctor Rene Halm. Madeleine Bastide was its President for several years. The group comprises actually more than 100 researchers from different disciplines (physicians, chemists, biologists, pharmacologists, physicists, etc. ) from 22 different countries. The original aim, which, for the most part, has now been achieved, was to create a working group to exchange ideas and results concerning the effects of very low doses and high dilutions. Madeleine Bastide then proposed to publish a collection of the lectures given at their scientific meetings in a bilingual book (French and English) entitled "Signals and Images." The first volume was published in 1990 by the ATELIERS ALPHA BLEUE. It contains lectures on the new pharmacological approach and related concepts that were presented at the 3rd and 4th Symposiums held in Paris in 1989 and 1990. This volume treats the problem of the effects of very low doses and high dilution in in vitro and in vivo experimental models. It is conceivable that this effect, like that of elect- magnetic fields, is none other than an "ultra molecular" effect as shown in yet p- lished results. The effect could be interpreted as a piece of information, i. e. a signal whose transmission and perception remains to be elucidated. According to Madeleine Bastide "everything depends on the nature and quality of the information, signals and images.""

As editor I want especially to thank Dr. Ernst Freese for helpful co operation in preparing these volumes, and to express my appreclatlOn to Drs. Kurt Hirschhorn and Marvin Legator, the other members of the editorial board. Alexander Hollaender January 1971 Preface The purpose of these volumes is to encourage the development and ap plication of testing and monitoring procedures to avert significant human exposure to mutagenic agents. The need for protection against exposure to possibly mutagenic chemicals is only now coming to be generally realized. The recently issued Report of the Secretary's Commission on Pesticides and Their Possible Effects on Health (the Mrak Report-U.S. Department of Health, Education and Welfare, December 1969) has made an important start. Its Panel on Mutagenicity recommends that all currently used pesticides be tested for mutagenicity in several recently developed and relatively simple systems. Whether recommendations such as these are actually put into effect will depend on convincing government, industry, and the public that the problem is important, that the proposed tests would be effective, and that they can be conducted at a cost that is not prohibitive. Why is it important to screen environmental agents for mutagenic activity? To those who will read this book, the answer is self-evident. The sine qua non of all that we value and all that we are is our genetic heritage.

This book grew out of an effort to salvage a potentially useful idea for greatly simplifying traditional quantitative risk assessments of the human health consequences of using antibiotics in food animals. In 2001, the United States FDA's Center for Veterinary Medicine (CVM) (FDA-CVM, 2001) published a risk assessment model for potential adverse human health consequences of using a certain class of antibiotics, fluoroquinolones, to treat flocks of chickens with fatal respiratory disease caused by infectious bacteria. CVM's concern was that fluoroquinolones are also used in human medicine, raising the possibility that fluoroquinolone-resistant strains of bacteria selected by use of fluoroquinolones in chickens might infect humans and then prove resistant to treatment with human medicines in the same class of antibiotics, such as ciprofloxacin. As a foundation for its risk assessment model, CVM proposed a dramatically simple approach that skipped many of the steps in traditional risk assessment. The basic idea was to assume that human health risks were directly proportional to some suitably defined exposure metric. In symbols: Risk = K x Exposure, where "Exposure" would be defined in terms of a metric such as total production of chicken contaminated with fluoroquinolone-resistant bacteria that might cause human illnesses, and "Risk" would describe the expected number of cases per year of human illness due to fluoroquinolone-resistant bacterial infections caused by chicken and treated with fluoroquinolones."

This volume on enzootic bovine leukosis (EBL) and bovine leukemia virus (BLV) is the second in our series "Developments in Veterinary Virology". Each book in this series is devoted to a major virus disease of agricultural significance. The chapters in each volume are planned to supply information on a range of subjects from pathogenesis of the causative virus to vaccination, eradication, and rules regarding disease control. The present volume on enzootic bovine leukosis and bovine leukemia virus updates the reader on the disease and its causative agent and includes the nucleotide sequence of the BLV genome as well as data on its integration into the DNA of the tumor cell. Insights into diagnosis, veterinary legislation, and the economic aspects of EBL are also provided. Intense research conducted on EBL and BLV during the course of a decade is presented in a most concise and in-depth manner, so as to provide the reader with a comprehensive overview of this economically important disease of cattle. I wish to thank the editors, A. Burny and M. Mammerickx, as well as all the authors, for making this excellent book available at a stage when the knowledge on bovine leukemia virus will also contribute to our understanding of the virus causing human AIDS.

The purpose of this book is to provide the reader with some basic information applicable to cattle feeding. It is intended to adapt some of the basic principles of nutrition in applied form. During the past few decades there have been various changes in type and form of feeds available for livestock feeding due to new kinds of equipment. Mechanization has made it possible to perform certain opera tions of the beef production program more efficiently and economically. With all the new innovations and advances in animal nutrition combined with the capabilities of the computer, it becomes very challenging for everyone to keep up to date on the latest information in the field of cattle feeding and production. The text was written with the intent of utilizing the raw materials, facilities, equipment, etc. which are available in the United States. The terminology of certain materials such as feed ingredients will vary from one country to another. One term which is frequently used in this text is forage. Although the term roughage is used more commonly in the United States it has been replaced with forage in this text. J.K. MATSUSHIMA Fort Collins, January 1979 Contents Chapter 1 Nutrients 1 Proximate Feed Analysis 1 Chemical Classification of Nutrients 2 1.1 Water 3 1.1.1 Drinking Water .... ...."

This book contains the contributions to a workshop on stress in farm animals held on April 17-18,1986 at the Pietersberg Con ference Centre Oosterbeek. The workshop was financed by the Commis sion of the European Communities from its budget for the coordina tion of Community Agricultural Research and the Agricultural Uni versity of Wageningen (The Netherlands). Its aim was to bring together experts from different disciplines all having in common that in one way or another they were involved in stress research. Such a multidisciplinary encounter should not only provide an interesting description of present day knowledge on stress, but also promote a more integrated view on stress phenomena as they occur in higher vertebrates. In the course of this workshop the following fields of research were related to stress: endocrinology, immunology, pathology, neurobiology, ethology and theoretical bio logy. Each of these relationships was introduce& by one speaker presenting a concise state of the art. The same relationship was elaborated by a second speaker implementing the available knowledge as far as possible to the farm animal situation. Therefore in this book each discipline is represented by a duo and introduced by some integrating remarks.

Reproductive wastage is a major inefficiency in all livestock production with embryonic mortality accounting for a major portion of this loss. Accordingly the Commission of the European Communities encouraged the organisation of a seminar on embryonic mortality in farm animals which was held in Brussels on the 11th and 12th of December 1984. This book contains the text of the papers, discussions and final summary presented at that Seminar. As a background to the Seminar, the extent and timing of embryonic loss was described for farm animals. Particular consideration was then given to the various mechanisms and signals, both embryonic and uterine in origin, that are so far known to be involved in the establishment of pregnancy. Possible causes of embryonic death including physiological, endocrinological, genetic and immunological components were outlined and discussed. The final summary contains general conclusions from the Seminar and recommendations for future research work on this topic. J.M. Sreenan M.G. Diskin July 1985. ******************** THE EXTENT AND TIMING OF EMBRYONIC MORTALITY IN THE COW J. M. Sreenan & M. G. Diskin, The Agricultural Institute, Belclare, Tuam, Galway, Ireland ABSTRACT The extent and timing of embryonic mortality in heifers, normal cows and repeat breeder cows has been reviewed.

Contrary to common belief, infectious diseases are not as well under control as we would like. We are now at a crossroads regarding the impact of the environment on infectious diseases. Renewed interest in biological weapons and the emergence of new pathogens, coupled with a better understanding of the impact of infectious agents on other conventional diseases, has led us to realise that we can no longer remain complacent about the impact of infectious agents on human, animal and crop health. The present book first discusses current and emerging military and civilian policies on the environment. In addition, the impact of environmental biology on the future of space exploration is discussed, especially in reference to the Mars mission. There follows a discussion of the state of bacteria in the environment, with a presentation of current and emerging techniques of microbial investigation. Finally, two case studies are presented on the impact of these techniques on both political and environmental problems.

The most rewarding aspect of writing a book is receiving encouraging comments from one's colleagues, since one always wonders whether fair coverage was made of the work of others or whether some omissions were made. I feel very fortunate that many colleagues took the time to read the first edition of this book and chose to use it as a textbook in their teaching. During the past few years they have given me valuable suggestions by pointing out areas that needed to be added to improve the book. Toxicology is one of the fastest moving scientific fields. In the areas of insecticide toxicology many new advances have been made since this treatise first appeared. Therefore, it would not be easy to write even a review paper that would not be outdated by the time it was published. In revising this volume I have made a conscious effort to adhere to the basic principles which have been developed over the years. While I have retained the basic framework of the original book, advances that fundamentally change certain concepts or add a new horizon have been chosen for updating those fields where applicable. The main emphasis has been placed on the addition of new sections and new compounds developed since 1975. Since the first edition appeared, several books covering technical details in each group have been published. They are excellent encyclopedic resources in their chosen areas, and are listed in each section as recommended reading material.

"Complementary and Alternative Veterinary Medicine Considered" is a book that belongs in your veterinary library. If you are a veterinarian wondering if you should incorporate complementary and alternative veterinary medicine (CAVM) into your practice, if you have recently hired an associate eager to try such things as acupuncture or homeopathy, or if you have clients asking you about chiropractic, herbal, or magnetic field therapy for their pets, you'll want to understand the history, science and ethics behind such therapies.

In its 2001 "Guidelines for Complementary and Alternative Medicine," the American Veterinary Medical Association (AVMA) recognizes the growing interest in CAVM, and encourages the critical examination of these therapies using the scientific method. Following the AVMA's lead on this subject, "Complementary and Alternative Veterinary Medicine Considered" thoroughly examines a variety of CAVM therapies and asks important questions regarding alternative treatments. For example, is acupuncture effective in pain relief? What is homeopathy? What is the history behind chiropractic? What does the research say (and not say) about various CAVM modalities? And, just as importantly, what are the ethical and regulatory considerations concerning such therapies? This book has the answers to those questions and more.

"Complementary and Alternative Veterinary Medicine Considered" will help practicing veterinarians to make informed decisions about specific CAVM therapies. This text evaluates various prevalent therapies, and will give veterinarians the ethical and scientific bases they need to make sound decisions regarding CAVM therapies

Coverage includes but isnot limited to:
Acupuncture and acupressure;
Energy medicine;
Manual therapy (chiropractic);
Manual therapy (massage);
Magnetic and electromagnetic therapy;
Laser and light therapy;
Homeopathy; and
Herbal therapy.

The use of ultrasonic imaging techniques for diagnostic purposes in veteri nary medicine and animal science has lagged far behind their use in human medicine. In the area of domestic animal reproduction, diagnostic ultrasono graphy has a relatively short history. Reports on B-mode scanning first appeared as late as 1969, when Stouffer and co-workers used it for counting foetal numbers in late gestation sheep. After Lindahl had re-evaluated the potential of two-dimensional ultrasonography for pregnancy diagnosis in sheep in 1976, results of the first large scale field-trials by Fowler and Wilkins on predicting foetal numbers during the first half of gestation in sheep were published as recently as 1980. In this year also the first paper, by Palmer and co-workers, on ultrasonic scanning of the uterus and ovaries in the mare appeared. The improvement of diagnostics which was achieved by the application of this technique is illustrated by its present routine use in clinical equine practice and sheep breeding. Besides offe ring an accurate early pregnancy diagnosis in species such as the horse, cow, sheep, goat, pig and dog, ultrasonography enables the visualization of ovarian and uterine structures for the identification of both physiological and pathological conditions. In this way it replaces or supplements the more common diagnostic techniques used by the general practitioner so far. Ultrasonography has also opened several new lines of research for clini cians and research workers."