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    Mario Manuel Fiorentino Ferreyros
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    CHAPTER Ethical Aspects of the Use of Animals in Experimental Research Vera Baumans Met has long used animals for food, for transport and as companions. The use of animals in experimental research parallels the development of medicine, which had its roots in ancient Greece. Aristotle and Hippocrates laid down their knowledge on structure and function of the human body in their respective Historia Animalium and Corpus Hippocraticum, mainly based on dissections in animals. Galen (130-201 AD), physician of the Roman emperor Marcus Aurelius, performed physiological experiments on pigs, monkeys and dogs; these experiments provided the basis for medical practices in the centuries thereafter. After Galen, experimental science remained in a dormant stage until the beginning of the Renaissance when Vesalius popularized the empirical approach, starting with anatomical studies, Later on, physiological studies were performed as well. With the advent of Cartesian philosophy in the 17" century, experiments on animals could be performed without great moral problems. The French philosopher Rene Descartes (1596-1650) stated that living systems could be understood on pure mechani- cal principles. The difference between man and animals is that man has a mind, which is a prerequisite for awareness and consequently for the capability of feeling pain. Animals cannot think and are more like machines. However, Jeremy Bentham (1789) opposed Descartes’ views: “The question is not, can they reason? Nor, can they talk? But can they suffer?” The discovery of anesthetics and Darwin's publication on the Origin of Species in 1859, defending the biological similarities between man and animal, contributed to an increase in animal experimentation. Claude Bernard published his book “Introduction 4 Pétude de la médecine expérimentale” in 1865, introducing methodology as a tool for the design of physiological experiments. The development of microbiology caused an in- crease in the use of animals, due to Koch's ‘Postulates’ where it is stated that the pathoge- nicity of a microorganism can be proven after successfully infecting healthy, susceptible 7 CHAPTER 2 animals (Van Zutphen, 2001). The development of biomedical disciplines such as phar- macology, toxicology and immunology caused a sharp increase in the use of animals in the 20" century. Since the early 1980's, animal experimentation has decreased due to public awareness; strict legislation regarding animal use; the development of animal eth- ics committees and improved animal quality (Figure 1). However, in recent decades the use of animals started to increase again, mainly due to the development of genetically modified animals, which has caused a 23% increase in the numbers of mice used each year. This increase reflects not only the animals used in research but also the large num- ber of mice necessary to create each genetically modified line (breeding males, donor females, vasectomized males and pseudo-pregnant recipient females), Furthermore, non- transgenic and wild-type littermates may be produced that are not suitable for research or further breeding (Dennis, 2002). Genetically engineered or modified mice are those with induced mutations, including mice with: transgenes; targeted mutations (knock- outs); and retroviral, proviral or chemically induced mutations. Transgenic technology focuses on the introduction or exclusion (knockout) of functional genetic material in the germ-line of an animal, thus changing the genetic characteristics of an organism and its progeny. These techniques have led to the rapid development of a variety of animal mod- els, designed for the study of gene regulation, gene expression, pathogenesis and the treat ment of human and animal diseases (¢.g., Alzheimer’s disease, growth hormone distur- bances, mastitis in cows, and poliovirus vaccine testing for eventual use in humans). genetic immunology: toxicology/one pharmacology Number («ml microbiology sg eee eeses Year FIGURE I = Development of animal use in the 20" century. ETHICAL ASPECIS OF THE USE OF ANIMALS IN EXPERIMENTAL RESEARCH ‘The experimental procedures involved in the process of transgenesis may compromise animal welfare, The donor animals, vasectomized males and foster mothers which are needed for the production of the transgenic offspring may experience discomfort from procedures, such as early mating (from 3 weeks onwards), anesthesia, surgery and injections. At the level of integration of the microinjected DNA into the genome, unintentional insertional mutations may occur, impacting the health of the animal. Furthermore, ex- pression of the introduced gene may cause detrimental side-effects, e.g,, the giant mouse with an overproduction of growth hormone, suffering from chronic kidney and liver dysfunction (Poole, 1995). ‘Transgenic technology has gteat potential for increasing our understanding of the role of genes and may provide suitable animal models for human and animal disease. However, the welfare of transgenic animals must be carefully monitored, at least until the second generation of offspring, Score sheets can be helpful in correlating animal health problems with humane end-points in order to euthanize severely affected animals (Crawley, 1998; Van der Meer et al., 2001). Data banks will be useful in providing data from existing genetically modified animals in order to predict possible health impair ments in newly created genetically modified lines. Today, 75-100 million vertebrates per year are used in research for a wide range of purposes, The major areas are drug research; testing of vaccines; and cancer research, whereas about 30% of the animals are used for other purposes such as fundamental re- search, and diagnostics (Figure 2). ‘Mice and rats are the most frequently used animal species (Figure 5). In many Euro- pean countries, itis mandatory by law to grade the level of discomfort experienced by the animals used in an experiment as minor, moderate and severe. On average, 50% of the laboratory animals experience minor discomfort (e.g., single blood sampling), 30% ex- perience moderate discomfort (e.g. recovery from anesthesia) and 20% experience se~ vere discomfort (e.g., toxicity tests). The increasing demand for high-quality animal models together with a critical view on the use of animals led to the development of Laboratory Animal Science in the fifties, a field that can be defined as a multidisciplinary branch of science, contributing to the quality of animal experiments and to the welfare of laboratory animals. Laboratory Ani- 10 CHAPTER 2 Cancer research 12% Heart/circul 2% Vaccines/biol 21% Drug research 23% Tusicitytests ducati % Other research 3 FIGURE 2 — Distribution of the purpo ‘mouse guinea pig 2% rabbit 1% other species fish birds 10% FIGURE 3 ~ Distribstion of vertebrate animal species used for research, testing and education. IL “Supyovay pur ‘sosodind ssuajap Arey 104 sysa1 ‘sysa1 gg CUT Suse} IAaWIsO9 UT se yINS spandie aq 1y8tur squaumadxa feuue pera Jo Aussadou ay) ‘soULDYZIN “(suOeUID -nyjey 40 soypepeay touur “8'2) sjeuue uy papayopun 08 Aeu 10 2ouapI9UT Mo} AjoUION -x9 0} anp paziuSooa1 aq 1ou deur sBnup Jo si2ay9 apis ayp ‘sueUMY UT speER yeDTUTD pue suaturiadxa jeurtue Jo sarias v auidsaq “uew 01 parejodennse Aqquyer aq you Stu syeur que ut paureygo synsay “paaruesend aq youu Aypigerr weyp asoud sey 410181} “SII -adxe jeuitue jo Ayissacau pur Agger ayy UO se [fam se ‘SpeUUTUe asn o2 IYBLI ayy sey, ‘ubur aatpaya Jo UORSaNb erp axp wo fexUas UT sosnd0j WIsIINLID ‘sasodind yoseIsar Jo} pasn a1am s[eUnue aoUIS 1A WHS!>HTID 0} 2a{qns Ud0q aavy stustUTIedxe [UII quoRwOosIp pur ured sazqenurur ye porpur e suo AuaMOUBO sjentur jo saquinu axp sonpaz :worsnpay, speumtue jo asn arp saovjdar quawiasejday, “Supsayns Aressaoouun quaaaad 03 paziueyina aq we> jeurrue ayy Uys uIod -pua sueumny e Suruyuaap Aq pue ssunpadoad peyuawutiadxe Suraordur Aq sSururen pue uoneonpe Ag paastyse aq URD YY Gers [eUMTUR/J9YDIvasas ay) JO S[[DYs ay) Survayuerens dq saze> pur eiso8peue ‘eisoyzsoue ayenbope Surpraoad dq ‘upuvgsny pur Sursnoy ayenbs -pe out jewtue ay Jo spaou jeorSoporskyd pur pesoraeyag ayy SuNPIsUEN Aq OJWO -sip Surseas.op surat UsMIOUyDY ‘sisAjeue Jomod “3:9 uautLiadxe ays 0} 101d sonstIEAS Sursn Aq pareumse Ajaremooe auout aq ues dno jeusuadxa aad sjeunue jo s9quunu ay sxouayNg ‘speuMUE ay Jo SuIaq-][2M axp aaordutt 04 9pso ut “Buysnoy Jepos pue quoUmpEue [eusUTONAta Jo sutsD) UT ‘spoau sypads-sapads s jewutUe ayy Bunga uA -voraue ue jo aaueuoduat 2yp paztuoras savy sioy>seasoa ‘apeoap ise] ayn SuLNG]“[eLAY -eui Surppog prepueys pue aBe> padeys-xoqaoys e soajoaut aonserd Asoresogey ‘JeNUassa posopistio> si ae [ewTUL ayp Jo VoNezprepuLIg “Woo [PUT ay UI aIeUET|D PUL Poo; Jo sua3) ut Apsejnon red ‘squauuotAua pur soanpasoud jeywaumtiodxa wuoyiun Sutsn sv jjan se Sayenb perrSoporqoxonu pue addrouas jo suisay ut eutue ayy Sutzrpavpurys dq posn sjeut -juv jo Jagumnu 24) Surses.vap sueow! UORINpay “sjapour poztainduto> 40 (sanssn ‘s|]92 $9) sonbruypor oa 1 BIN syeuntue Suray Jo asn ay SuNMUSGns sues yusuase [doy (6661 ‘young pur [jassmy) ,anbruqzay, eruoutadxy aueuMp Jo sajdioutig oy, Yooq 4194) UI 6S6I UI YANG pure [possny Aq paypuney quawoUYsY pur LONSNpay quatar2|doy JO SY ¢ ayp aue sajdourid Burp ayy ‘some pue swuaumtadxa jeunue o} ssaneuseye ‘erseuteapina pur ersa8pour “exsaxpsaute ‘sanbruypa ferttouutzadxo Jo wawoaosduut ‘soseasip. Jo qusWAeAN pur UoNUdsaid ‘oNezprepUrys erIFOJoIgo.D1U pue Indu ‘susWAZMb “au [PQUDWIUOIIAUA sTYp puL speusTUE AzoWeIOqGP] Jo ABojOIg axp sassedurosuia asUa!Ds JeUt HOUVaSIU TVLNANIEEXA NI STVWINY 40 98M BHA JO SIDTdSV WIHT 12 CHAPTER 2 Undoubtedly, activities of the general public such as animal protection organizations have contributed to legislative regulations for the protection of animals used for experi- mental purposes. The first law, the Cruelty to Animals Act, was adopted by the UK parlia- ‘ment in 1876, Since then, other countries have included provisions in their laws to pro- tect experimental animals. In Europe, two important documents controlling the use of animals in experiments were issued. In 1985, the Council of Europe (European Conven- tion for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes) signed the Convention for the Protection of Vertebrate Animals used for Experi- mental and other Scientific Purposes (ETS 123). In 2006, the EU passed a European Coun- il Directive, the Directive for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes (86/609/EEC), based on ETS 123, but more stringent. The Directive applies to vertebrate animals used in experiments likely to cause pain, suffering, distress or lasting harm, extending to the development of genetically modified animals at risk for pain and distress. The Directive contains provisions for the accommodation and care of experimental animals and for the competence of researchers and animal staff. It also lists alternatives to animal experiments, as well alternate forms of anesthesia and euthanasia. The document also contains statistics on animal experimentation and the supply of animals, In the US Animal Welfare Act (2002) the term “animal” means any live or dead dog, cat, monkey, guinea pig, hamster, rabbit, or other warm-blooded animal being used, or intended for use, for research, testing, experimentation, exhibition pur- poses, or as a pet. However, the term excludes birds, rats of the genus Rattus and mice of the genus Mus, bred for use in research. The guidelines for accommodation and care of laboratory animals are included in the Guide for the Care and Use of Laboratory Animals of the National Research Council, which also covers rats and mice (National Research Council, 1996). Although no specific provision in the EU Directive demands the establishment of animal ethics committees, such committees are operational in several European coun- tries. These groups are specifically dedicated to reviewing the ethical aspects of animal experimentation. In the US, comparable Institutional Animal Care and Use Committees (IACUC’s) are in operation. The task of such a committee is to perform an ethical evalu- ation of submitted research proposals. Animal experiments are considered acceptable only if the benefit of the proposed experiment outweighs the suffering of the animals. Ethical review of animal experiments will likely benefit the animals and improve the quality of animal-based research, As animal well-being is a prerequisite for reliable ex- perimental results; it is of the utmost importance to seck methods and procedures that can reduce the suffering of animals and improve their well-being. ETHICAL ASPECTS OF THE USE OF ANIMALS IN EXPERIMENTAL RESEARCH REFERENCES Crawley IN. What's wrong with my mouse? Behavioral phenotyping of transgenic and knvock- cut mice. New York, USA: Wiley-Liss; 1998, Dennis Je MB, Welfare issues of genetically modified animals. LAR } 2002;43(2):100-9. European Convention for the Protection of Vertebrate Animals used for Experimental and

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