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Methods of Animal Experimentation. Volume I. Book • Edited by: WILLIAM Contents of Volume II CHAPTER 7 - Methods in Germfree Animal Research.
Table of contents

The use of animals for scientific or teaching purposes should be considered only when there is no alternative. Ethical considerations in the use of animals are the basis of the authorization of projects.


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The application of the principles of replacement, reduction, and refinement must be guaranteed. The means are put in place so that the animals do not necessarily suffer, and they are provided with analgesics and anesthetics to minimize the suffering or anguish. Euthanasia methods appropriate to the animal species and the procedure performed are used. The personnel participating in the procedures have the appropriate training training and experience to carry out the tasks entrusted to them.

An experiment is a procedure for collecting scientific data in a systematic way to answer a question correctly or for the generation of new hypotheses. All research should be described in such a way that the study design could be repeated elsewhere [ 1 , 2 ] Table 1. Animal research has made major contributions to the health and welfare of humans and domestic animals. These and many other advances have enabled physicians to treat a wide range of human diseases.

Many experiments appear to be poorly designed and inadequately analyzed and reported.


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As a result, some are found to be unrepeatable, leading to a waste of animals and scientific resources. Critical appraisal is an essential part of the scientific process designed to assess the validity of scientific findings. The new techniques of systematic reviews and meta-analyses are hampered by poorly written papers. The importance of randomization and blinding does not always seem to be understood, and it seems that many scientists have inadequate training in experimental design and statistics [ 7 ].

Animal studies differ from clinical studies in some aspects, such as the diversity of animal species studied, experimental design, and study characteristics.

Part 2: Politics and Legislation of Animal Experimentation

These methods used in animal studies are explained in [ 8 ]. For ethical and economic reasons, it is important to design animal experiments well, to analyze the data correctly, and to use the minimum number of animals necessary to achieve the scientific objectives—but not so few as to miss biologically important effects or require unnecessary repetition of experiments [ 4 ].

The 3Rs—replacement, reduction, and refinement—can be applied to any animal experiment by researchers and other bodies seeking to conduct those studies in as humane manner as possible. Key to the success of this endeavor is an appreciation of the principles of good experimental design and analysis; these need to be considered in concert before any data is collected and understanding of animal welfare plays a central role in laboratory practice—are to the betterment of research per se [ 40 ].

Careful choice of the animal model is essential, if research is to be conducted efficiently, by using the minimum number of animals in order to provide the maximum amount of information. Inbred strains of rodents provide an excellent way of controlling and investigating genetic variation in characters of interest and in response to experimental treatments. Outbred stocks, in which genetic and nongenetic factors are inextricably mixed, are much less suitable, because random and uncontrolled genetic variation tends to obscure any treatment responses [ 10 ].

There is concern about the lack of repeatability of many preclinical experiments involving animal toxicity tests in rodents used to assess the safety of drugs to detect adverse effects that have not been formally evaluated. However, the test does not specify the strain of animals in which the genetic variation, is unknown and uncontrolled; a better strategy would be to use small numbers of animals of several genetically defined strains of mice or rats instead of the undefined animals used in the present.

Inbred strains are more stable providing more repeatable data than outbred stocks [ 11 ]. One of the uses of animal models is related to the evaluation of new drugs for the treatment of human diseases.

Description

For this type of use, the animal model must respond adequately to the effects of different therapeutic agents. Some of this may be due to poor design of the animal studies. But in some cases, the animal model may not be truly representing the human condition.

Research using animals: an overview

It is suggested that a good model of a human disease should also have the same human biomarkers of that disease [ 7 ]. Compounds that are active in routine clinical practice should show activity in the model positive controls , and compounds that show no activity in clinical practice should not show effects in the animal model negative controls [ 12 ]. Most animal laboratory models have been developed and used for the study of the cause, nature, and cure of diseases in humans.

There are five categories of experimental models, of which the first three are the important ones, since they are the most used: Induced animal models. The selection of any animal model for research should be based on the following considerations: models based on analogy similar structures involve similar functions and models based on homology structures derived from the same evolutionary precursor have the same or similar functions. The most appropriate selection of an animal species for the experimental purpose should not be based on its easy management due to its small size, availability, familiarity, or cost [ 13 ].

However, scientists recognize that there are no real substitutes in the use of laboratory animals. Studies with bacteria, tissue cultures, and computer simulations can provide useful information, but the complexity of living organisms requires research and analysis on animals similar to humans to achieve reliable results. When considering which can be the best animal model to use, it is important to take into account the extrapolation or generalization of results that this model generates. It is only possible in the case that in the animal model, the respective injuries to the associated syndromes resemble each other.

Isomorphic models: similar symptoms but the cause does not have to be the same. For example, in a neural zone degeneration pathology, we can alter that same area in rat brain and see that the symptoms are identical. Partial: Some of the models do not completely imitate the human disease, but they can be used in the study of certain aspects or treatments of the human disease, considering that an optimal model would be one that develops a comparable symptomatology, etiology, and neurophysiological background and that responds similarly to the effects of different therapeutic agents.

To designing any scientific investigation once, having an idea for a research project is necessary to make a review of the literature and to get the information that is necessary for the experimental design phase. A null and an alternate hypotheses that address the problem statement are then formulated, and only then is the specific design of the experiment developed. The identification of the most appropriate animal model to address the experimental question being asked is very important.

Animal Handling and Experimentation

Other aspects are the considerations that include the number of animals needed per group and evaluating the most appropriate statistical analyses [ 14 ]. Nowadays models of human diseases are necessary for experimental research into the biological basis of disease and for the development of treatments. They have an enormous impact upon the success of biomedical research. However, in spite of this, a consistent system for evaluating, expressing, and comparing the clinical validity of disease models is not available [ 14 ].

Usually, studies are performed on animal species such as genetically heterogeneous GH mice, and rats continue to be used in research even though the case for using isogenic strains has been argued repeatedly. GH stocks represent poor material for controlled studies because genetic heterogeneity normally leads to phenotypic variability and a decline in experimental sensitivity.

Isogenic strains are a vital, proven, and powerful resource for biomedical research and should be used in preference to GH stocks by all scientists who use laboratory rodents [ 15 ]. It is impossible to give specific rules for the selection of the best animal model; however, it is convenient to make many considerations before an experiment. These are some general rules regarding the criteria for choosing the model [ 16 ] Table 2. It is also important to identify in usual practice among other criteria for the selection or rejection of a model the presence of diseases or special conditions of the animal and that the microbiological status of animal can influence their response [ 13 ].

These factors should be considered when choosing the animal model that best suits the experimental purpose. Many models that do not use animals have also been developed, refined, and characterized. These models are useful in some types of research and testing, and they can often be used to complement work with live animals. Animals have been used in research and teaching for a long time. However, ethical guidelines and pertinent legislation were instated only in the past few decades; even in developed countries guidelines for animal experimentation vary.

The EPAA is one of the leading organizations in Europe for the promotion of alternative approaches to animal testing [ 18 ]. The alternative methods are based on the principle of the 3Rs [ 19 ] established by Russell and Burch in R of reduction, using only the number of animals needed to obtain a reliable and accurate information; R for refinement understood as any system that allows to reduce the severity of the damage inflicted on the animals; and R for the replacement of vertebrates by any other method that uses nonsensitive material. All methods or techniques that could substitute the experiments carried out with animals, reduce the number of animals used in each trial, or improve existing procedures in order to reduce stress and avoid the suffering of the animals that are included.

The principle of the 3Rs has been responsible, in large part, for the drastic reductions in the use of laboratory animals that have occurred in the last century and for the significant changes in the techniques of research, testing, and education for the benefit of science and public health, as well as animals.

Undoubtedly, the promotion of alternative approaches is one of the basic aspects that permeate the new animal protection regulations. Experimental alternative methods include any procedure that replaces the use of animals, that reduces the need for animals in a particular test, or that refines a technique in order to reduce the amount of suffering endured by the animal. To be used in the toxicity tests required for the register prior to the commercialization, transportation, and use of a new chemical compound, it is necessary for the experimental procedure to be accepted by regulatory authorities.

The principles of good laboratory practice GLP are designed to help ensure the proper management and conduct of studies. GLP compliance demonstrates to regulatory authorities that studies were undertaken in a manner which promotes confidence in the data and reporting. Study management of interlaboratory studies in compliance with GLP is discussed [ 21 ]. The alternative approaches undoubtedly provide alternatives available to animal research to raise awareness of viable and, at times, even better options outside of animal experimentation. They currently cannot replace animal testing altogether.

The extant alternatives serve to complement animal experimentation in current research [ 17 ]. Basic animal handling, anesthetization, blood collection, drug administration, and euthanasia. The development of alternative methods for teaching is not new, and so in the report of the meeting of experts in alternative methods in teaching, organized by ECVAM in [ 24 , 39 ], several types of methods were already identified:. There are several modalities of alternatives that can be used in teaching [ 22 , 23 , 24 ] Table 3.

If an adequate system is not located, the bibliographic databases could also be revised. There are also systems aimed at improving the preparation of people who handle experimental animals.

Research using animals: an overview | University of Oxford

The mechanical models consist of reproductions of animals or organs that allow training in management techniques, administration, extraction, and surgery. The classical audiovisual systems were the first used to show the techniques of animal handling, to learn comparative anatomy and various specific techniques. The OOC looks promising as a pathophysiologically pertinent model of experimentation.

In vitro models of skin pathophysiology and drug testing have been around for some time. Presently, HSE models are used to demonstrate simple physiology, to analyze autoimmune disorders to malignancies [ 29 , 30 ]. These models may be better than animal models because the skin samples are human-derived.