**Acknowledgements**

My first impression to ponder upon the link between 3D technology and its medical application happened in a collaboration research discussion conducted with colleagues at the University of Gondar in Ethiopia in early 2017. Hence, my thanks goes to the University of Gondar for providing a platform for the meeting of experts in the field of law, technology and human rights. My special thanks goes to my friend and colleague, Dr. Hailemichael Demissie, who first tabled the importance of 3D printing technology for harnessing Africa's development, on the basis of which I personally appreciated the significance of linking 3D printing as a useful avenue for promoting the right to healthcare in Africa.

## **Author details**

Solomon Tekle Abegaz

Address all correspondence to: solomomte@gmail.com

School of Law, University of Gondar, Gondar, Ethiopia

## **References**


liberalization and deregulation to ensure a more central, dynamic and effective contribution of communication to the work of the Commission. Driven by technological convergence, it is here argued that the concentrated use of 3D technology can bring unprecedented comparative advantages to the continent. The knowledge-based economy of the future will depend more and more on the effective use of this technology. Rapid advances in technology coupled with the low-cost of acquiring 3D technology tools are opening new windows of opportunity for Africa to accelerate access to healthcare services. The 3D printing revolution can accelerate Africa's goals in the right to health, fostering intra-regional trade, integration into the global economy, as well as realizing its security needs. This must be reinforced by African governments' political will to improving knowledge, skills and resources and creating collaboration

My first impression to ponder upon the link between 3D technology and its medical application happened in a collaboration research discussion conducted with colleagues at the University of Gondar in Ethiopia in early 2017. Hence, my thanks goes to the University of Gondar for providing a platform for the meeting of experts in the field of law, technology and human rights. My special thanks goes to my friend and colleague, Dr. Hailemichael Demissie, who first tabled the importance of 3D printing technology for harnessing Africa's development, on the basis of which I personally appreciated the significance of linking 3D printing as

[1] Jennifer MM. Human rights and development: Using advanced technology to promote human rights in sub-Saharan Africa. Case Western Reserve Journal of International Law.

[3] UN Committee on Economic, Social and Cultural Rights. General comment no. 14. The Right to the Highest Attainable Standard of Health. Article 12 of the Covenant. 11

[2] World Health Organization (WHO). Constitution of the WHO. 1946

and consensus among key stakeholders.

a useful avenue for promoting the right to healthcare in Africa.

Address all correspondence to: solomomte@gmail.com School of Law, University of Gondar, Gondar, Ethiopia

**Acknowledgements**

132 Reflections on Bioethics

**Author details**

**References**

Solomon Tekle Abegaz

1998;**30**(343):343-371

August 2000. E/C.12/2000/4


[20] June J. Dubai Is Going to be Home to the World's First 3D Printed Skyscraper [Internet]. March 16, 2017. Available from: https://futurism.com/dubai-is-going-to-be-home-to-theworlds-first-3d-printed-skyscraper/ [Accessed January 3, 2018]

[34] Human Rights Watch. Digital Disruption of Human Rights [Internet]. March 25, 2016. Available from: https://www.hrw.org/news/2016/03/25/digital-disruption-human-rights

Marching for 3D Printing: Its Potential to Promoting Access to Healthcare in Africa

http://dx.doi.org/10.5772/intechopen.75649

135

[35] Yvonne D. The right to enjoy the benefits of scientific progress: In search of state obligations in relation to health. Medicine, Health Care, and Philosophy. 2011;(4):371-381.

[Accessed December 24, 2017]

DOI: 10.1007/s11019-011-9327-y


[34] Human Rights Watch. Digital Disruption of Human Rights [Internet]. March 25, 2016. Available from: https://www.hrw.org/news/2016/03/25/digital-disruption-human-rights [Accessed December 24, 2017]

[20] June J. Dubai Is Going to be Home to the World's First 3D Printed Skyscraper [Internet]. March 16, 2017. Available from: https://futurism.com/dubai-is-going-to-be-home-to-the-

[21] Schubert V, Langeveld MCC, Donoso LA. Innovations in 3D printing: A 3D overview from optics to organs. The British Journal of Ophthalmology. 2014;**98**:159-161. DOI:

[22] Lu YGT, Wang MY. 3D printing and neurosurgery: Ready for prime time? World

[23] Permissionless Innovation. Advanced Medical Technology [Internet]. Available from: http://permissionlessinnovation.org/advanced-medical-technology/ [Accessed

[24] Banks J. Adding value in additive manufacturing: Researchers in the United Kingdom and Europe look to 3D printing for customization. IEEE Pulse. 2013;**4**:22-26. DOI:

[25] Stratasys. Advancing Health Care with 3D Printing: Applications and guidance on mate-

[26] Alicia M. The Evolution of 3D Printing: Past, Present and Future [Internet]. August 01, 2016. Available from: https://3dprintingindustry.com/news/evolution-3d-printing-past-

[27] Sunday AA, Clifford O, Olusina B, Opeyemi F. Barriers to accessing health care in Nigeria: Implications for child survival. Global Health Action. 2014;**7**:1-10. DOI: 10.3402/

[28] Solomon TA. Access to healthcare facilities: Promises and challenges in Ethiopia [Internet]. The Reporter. November 2013. Available from: http://www.thereporterethio-

[29] Sulaiman P. Africa can improve access to healthcare for all [Internet]. May 08, 2017. Available from: https://www.brandsouthafrica.com/play-your-part-category/play-yourpart-news/other-news/africa-can-improve-access-healthcare+&cd=5&hl=en&ct=clnk

[31] Convention on the Elimination of All Forms of Discrimination against Women.

[32] Convention on the Rights of Persons with Disabilities. May 3, 2008. General Assembly

[33] International Convention on the Elimination of All Forms of Racial Discrimination.

[30] Council of Europe, European Social Charter (Revised). May 3, 1996. ETS 163

worlds-first-3d-printed-skyscraper/ [Accessed January 3, 2018]

Neurosurgery. 2013;**80**:233-235. DOI: 10.1016/j.wneu.2013.07.009

present-future-90605/ [Accessed December 21, 2017]

September 1981. General Assembly resolution 34/180

January 4, 1969. General Assembly resolution 2106 A (XX)

10.1136/bjophthalmol-2013-304446

December 13, 2017]

134 Reflections on Bioethics

gha.v7.23499

10.1109/MPUL.2013.2279617

rial selection. 2016. pp. 1-34

pia.com/ [Accessed July 12, 2017]

[Accessed January 23, 2018]

resolution 61/106

[35] Yvonne D. The right to enjoy the benefits of scientific progress: In search of state obligations in relation to health. Medicine, Health Care, and Philosophy. 2011;(4):371-381. DOI: 10.1007/s11019-011-9327-y

**Section 4**

**Bioethics in the Use of Experimental Animals**

**Bioethics in the Use of Experimental Animals**

**Chapter 8**

**Provisional chapter**

**Bioethics in the Use of Experimental Animals**

**Bioethics in the Use of Experimental Animals**

DOI: 10.5772/intechopen.74520

This chapter deals with the history of the humanitarian use of animals in laboratory experiments from ancient times to the present day. It emphasizes the various criteria that have been established to try to improve the quality of life of an animal and its sacrifice with euthanasic techniques, since the emergence of Russell's statement of the three Rs (replacement, reduction, and refinancing). In addition, there is a review of the application of bioethical principles in scientific institutions in developing countries, such as Mexico. It also reviews some aspects of the humanitarian treatment of experimental animals at

"The greatness of a nation and its moral progress can be judged by the way its animals are

Since many centuries ago, the human being understood that animals, in addition to providing companionship, food and protection, could also be a source of knowledge. In this way, indiscriminate use has been made of many animal species throughout civilizations. Different species, including humans, have served to enhance the well-being and the art of human science, but it was not until the twentieth century, when it began to prohibit experimentation with humans, to use species phylogenetically very close to humans with scientific purposes; this was without considering how to design experiments and without taking into account the animal suffering infringed on them. Moreover, it was not until the mid-twentieth century that

**Keywords:** bioethics, laboratory animals, experimental design, three Rs,

© 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

© 2018 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use,

distribution, and reproduction in any medium, provided the original work is properly cited.

Tomás Alejandro Fregoso Aguilar and

Tomás Alejandro Fregoso Aguilar and

Additional information is available at the end of the chapter

Additional information is available at the end of the chapter

the time of designing an experiment protocol.

Elizabeth Guarneros Bañuelos

Elizabeth Guarneros Bañuelos

http://dx.doi.org/10.5772/intechopen.74520

**Abstract**

bioethics in Mexico

treated," Mahatma Gandhi.

**1. Introduction**

#### **Bioethics in the Use of Experimental Animals Bioethics in the Use of Experimental Animals**

DOI: 10.5772/intechopen.74520

Tomás Alejandro Fregoso Aguilar and Elizabeth Guarneros Bañuelos Tomás Alejandro Fregoso Aguilar and Elizabeth Guarneros Bañuelos

Additional information is available at the end of the chapter Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/intechopen.74520

**Abstract**

This chapter deals with the history of the humanitarian use of animals in laboratory experiments from ancient times to the present day. It emphasizes the various criteria that have been established to try to improve the quality of life of an animal and its sacrifice with euthanasic techniques, since the emergence of Russell's statement of the three Rs (replacement, reduction, and refinancing). In addition, there is a review of the application of bioethical principles in scientific institutions in developing countries, such as Mexico. It also reviews some aspects of the humanitarian treatment of experimental animals at the time of designing an experiment protocol.

**Keywords:** bioethics, laboratory animals, experimental design, three Rs, bioethics in Mexico

## **1. Introduction**

"The greatness of a nation and its moral progress can be judged by the way its animals are treated," Mahatma Gandhi.

Since many centuries ago, the human being understood that animals, in addition to providing companionship, food and protection, could also be a source of knowledge. In this way, indiscriminate use has been made of many animal species throughout civilizations. Different species, including humans, have served to enhance the well-being and the art of human science, but it was not until the twentieth century, when it began to prohibit experimentation with humans, to use species phylogenetically very close to humans with scientific purposes; this was without considering how to design experiments and without taking into account the animal suffering infringed on them. Moreover, it was not until the mid-twentieth century that

© 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. © 2018 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

some scientists began to consider designing experiments on animals, trying to cause as little suffering as possible and creating the first ethical committees for experimentation in science.

achievements went to the background. In the 1960s, movements for the rights of oppressed minorities appeared, which, using the same arguments toward animals, led to the famous animal liberation movements. Reaching its climax with the Australian philosopher Peter Singer (born in 1946), who wrote in 1975 his work "Animal Liberation" [6], Singer proposed an ethic that, starting from man, was also directed toward the rest of the animals. He attacked what he called "speciesism" or belief in the superiority of one species (the man) over others. A few years later, in several parts of the world, the idea of "animal rights" arose when in 1978 the "Universal Declaration of the Rights of Animals" was proclaimed by the UNESCO and the UNO [7], affirming among others things that animals have the right to: (1) live without hunger and thirst, (2) live comfortably, (3) live without suffering and illness, (4) express nor-

Bioethics in the Use of Experimental Animals http://dx.doi.org/10.5772/intechopen.74520 141

With the previous historical review, it could be inferred that currently there is an entire series of guidelines about ethical animal use in the laboratory, albeit they are not necessarily respected in all those countries where research is taking place on this matter or where subjects in relationship with the biomedical areas are taught. That is, in many places investigation and teaching remain using animals without the adequate measures (humanitarian) for their maintenance, handling and sacrifice. Talking about an ethic in the laboratory, animal handling looks like a utopic idea in many developing countries. Although some countries try to respect the international guidelines about animal experimentation, it is possible to claim that there is

It has understood that a laboratory animal is any animal species used for the purpose of

**2.** Biological reagent: the animal is considered like a biological substratum that can be put down to treatment in order to observe the result; this answer is reliable, duplicated and

**3.** Biological model: to extrapolate the results of a treatment from one specie to another, generally the human, with the purpose of improving the existing treatments [3, 9].

On the other hand, it is necessary to give a definition of animal experimentation too. In accordance with Mrad-De-Osorio and Rosenkranz [10], and Tobón-Marulanda [11], this concept refers to any experimental procedure that causes an alteration on the animal's well-being with the likelihood of causing it pain, suffering, anguish or discomfort. The objective of this procedure is always to make evident biological phenomena in that specie, even if these results are not compatible with human beings. The most complex designs are of the clinical type

**1.** Raw material: being exposed to different experimental variables, waiting for a result**;**

backwardness of many decades in comparison to the developed countries.

scientific experimentation. In this regard, a laboratory animal can be used as:

although there is no perfect model that can be extrapolated with humans.

mal behavior and (5) live without fear and anguish [5–8].

**3. Contemporaneous panorama**

comparable; and

When men were trying to know the why of biological processes and their pathologies in antiquity, the vivisection of men and animals was allowed alike; there are records that Persian physicians experimented with subjects condemned to death. In the time of Ptolemy, medical practice in criminals was allowed, reaching the point that Celso, in the second century, justified these practices saying that "it is not cruelty to inflict suffering on a few, when the benefit is for many" [1].

## **2. Historical review**

Since transition from nomadic to sedentary life, with the discovery of agriculture and the formation of the first settlements, prehistoric man became aware of the need to use animals to obtain meat, clothing and help in the transport of materials, thus emerging the basis of domestication.

It is in ancient Greece when a more "scientific" approach to the treatment of diseases with Galen and Hippocrates is obtained; however, it was not until the time of Andreas Vesalius, a doctor born in Belgium in the sixteenth century, when he changed the medicine, doing dissections of corpses of humans and animals. In the Middle Ages, great medical knowledge was obtained using animals, but in many cases, this knowledge was obtained by considering them as mere use and disuse objects, as René Descartes did, who claimed that animals had a lack of thought and conscience, concluding therefore that they did not have the capacity to feel pain. On the other hand, some scientific people of that time already began to think about the way in which studies were made in living beings; as an example to this, history has that in the works of Leonardo Da Vinci (1452–1519), he made contributions to the anatomy with dissections in dogs and cats but predicted that "one day, animal experimentation would be judged as a crime" [2]. It is in the seventeenth and eighteenth centuries, when men like Graff, Harvey, Malpighi, Aselli and Haller obtained physiological and histological knowledge from animal experimentation, in many cases, they were not anesthetized [3]. However, Schopenhauer (1788–1860) affirmed in his philosophical essays that animals were aware and could perceive pain. From this moment, currents of thought began to emerge that questioned the suffering of the animal in exchange for knowledge generation; in this sense, Jeremy Bentham (1748–1832) made clear that the questions were not: can they reason ?, can they talk ?, but rather: can they suffer? [3, 4].

In the second half of the nineteenth century, the Royal Society for the Prevention of Cruelty to Animals was founded in the United Kingdom and specifically in 1876, this country passed a law against cruelty to animals [4, 5].

Already entered the twentieth century, the English-speaking countries continued to set the guideline in terms of legislation in favor of the protection of animals, but it must be clarified that in the course of the two world wars (1914–1918 and 1939–1945), these issues and his achievements went to the background. In the 1960s, movements for the rights of oppressed minorities appeared, which, using the same arguments toward animals, led to the famous animal liberation movements. Reaching its climax with the Australian philosopher Peter Singer (born in 1946), who wrote in 1975 his work "Animal Liberation" [6], Singer proposed an ethic that, starting from man, was also directed toward the rest of the animals. He attacked what he called "speciesism" or belief in the superiority of one species (the man) over others. A few years later, in several parts of the world, the idea of "animal rights" arose when in 1978 the "Universal Declaration of the Rights of Animals" was proclaimed by the UNESCO and the UNO [7], affirming among others things that animals have the right to: (1) live without hunger and thirst, (2) live comfortably, (3) live without suffering and illness, (4) express normal behavior and (5) live without fear and anguish [5–8].

## **3. Contemporaneous panorama**

some scientists began to consider designing experiments on animals, trying to cause as little suffering as possible and creating the first ethical committees for experimentation in science. When men were trying to know the why of biological processes and their pathologies in antiquity, the vivisection of men and animals was allowed alike; there are records that Persian physicians experimented with subjects condemned to death. In the time of Ptolemy, medical practice in criminals was allowed, reaching the point that Celso, in the second century, justified these practices saying that "it is not cruelty to inflict suffering on a few, when the benefit

Since transition from nomadic to sedentary life, with the discovery of agriculture and the formation of the first settlements, prehistoric man became aware of the need to use animals to obtain meat, clothing and help in the transport of materials, thus emerging the basis of

It is in ancient Greece when a more "scientific" approach to the treatment of diseases with Galen and Hippocrates is obtained; however, it was not until the time of Andreas Vesalius, a doctor born in Belgium in the sixteenth century, when he changed the medicine, doing dissections of corpses of humans and animals. In the Middle Ages, great medical knowledge was obtained using animals, but in many cases, this knowledge was obtained by considering them as mere use and disuse objects, as René Descartes did, who claimed that animals had a lack of thought and conscience, concluding therefore that they did not have the capacity to feel pain. On the other hand, some scientific people of that time already began to think about the way in which studies were made in living beings; as an example to this, history has that in the works of Leonardo Da Vinci (1452–1519), he made contributions to the anatomy with dissections in dogs and cats but predicted that "one day, animal experimentation would be judged as a crime" [2]. It is in the seventeenth and eighteenth centuries, when men like Graff, Harvey, Malpighi, Aselli and Haller obtained physiological and histological knowledge from animal experimentation, in many cases, they were not anesthetized [3]. However, Schopenhauer (1788–1860) affirmed in his philosophical essays that animals were aware and could perceive pain. From this moment, currents of thought began to emerge that questioned the suffering of the animal in exchange for knowledge generation; in this sense, Jeremy Bentham (1748–1832) made clear that the questions were not: can they reason ?, can

In the second half of the nineteenth century, the Royal Society for the Prevention of Cruelty to Animals was founded in the United Kingdom and specifically in 1876, this country passed a

Already entered the twentieth century, the English-speaking countries continued to set the guideline in terms of legislation in favor of the protection of animals, but it must be clarified that in the course of the two world wars (1914–1918 and 1939–1945), these issues and his

is for many" [1].

140 Reflections on Bioethics

domestication.

**2. Historical review**

they talk ?, but rather: can they suffer? [3, 4].

law against cruelty to animals [4, 5].

With the previous historical review, it could be inferred that currently there is an entire series of guidelines about ethical animal use in the laboratory, albeit they are not necessarily respected in all those countries where research is taking place on this matter or where subjects in relationship with the biomedical areas are taught. That is, in many places investigation and teaching remain using animals without the adequate measures (humanitarian) for their maintenance, handling and sacrifice. Talking about an ethic in the laboratory, animal handling looks like a utopic idea in many developing countries. Although some countries try to respect the international guidelines about animal experimentation, it is possible to claim that there is backwardness of many decades in comparison to the developed countries.

It has understood that a laboratory animal is any animal species used for the purpose of scientific experimentation. In this regard, a laboratory animal can be used as:


On the other hand, it is necessary to give a definition of animal experimentation too. In accordance with Mrad-De-Osorio and Rosenkranz [10], and Tobón-Marulanda [11], this concept refers to any experimental procedure that causes an alteration on the animal's well-being with the likelihood of causing it pain, suffering, anguish or discomfort. The objective of this procedure is always to make evident biological phenomena in that specie, even if these results are not compatible with human beings. The most complex designs are of the clinical type although there is no perfect model that can be extrapolated with humans.

As the reader will understand, the concepts mentioned above establish some criteria of laboratory animal handling, but it does not classify those considering ethical aspects about the use of species in laboratory. For this reason, in many cases, the ethical aspects about the inclusion of animal experimentation depend on the exclusive experimenter judgment, who designs the treatment to obtain results in a short or medium term to use them for human treatment or, at least, to publish the researcher's scientific results in a scientific magazine which will be rewarded with "points" for the curriculum vitae or for receiving scholarships. This leads to keep ethical aspects aside at the point of making an experiment design. This leads to putting aside the ethical aspects when designing an experiment, to which is added that in countries like Mexico, in many of the institutions where research is done, just a few years ago, it has begun to integrate committees of bioethics that evaluate and ensure that animals are treated in accordance with international ethical statutes. On the other hand, problems can arise when submitting an article to be published in a scientific journal, when an author comes from a country where there are different ethical laws that do not follow these rules to the letter [12]. Even worse, in many countries, there is an infinity of laws for the ethical animal handling regulation and the author (with the editors) must decide which one of these to follow to write the article. As an example, in the United States, in 2004, 2100 laws for animal wellbeing were proposed [8]. However, in a certain way, in 2006, it was intended to be solved when the International Committee of Medical Journal Editors recommended the authors to report if animal experiments had been conducted according to the institutional and national guidelines about the use and care of laboratory animals [12]. In general, a laboratory animal should be kept in appropriate conditions, including its storage place, its feeding and its genetic characteristics (**Figure 1**).

Replacement: This includes conscious animals for unconscious animals or insensitive materials. To get this it is necessary to consider the use of *in vitro* systems, audiovisual aids, sacrificed animals, slaughterhouse material, use of invertebrates, human material, human volunteers or

Bioethics in the Use of Experimental Animals http://dx.doi.org/10.5772/intechopen.74520 143

Reduction: This includes reducing the number of animals without accuracy diminution. For this, it should consider the use of genetically homogeneous colonies without environmental influences; the animal model selected; sanitary, genetics and environmental quality; cryopreservation; advanced biostatistics methodology; data bank (publishing negative results to

Refinancing: It involves pain and discomfort reduction techniques taking into consideration animal care and well-being, dexterity and training of laboratory personnel to make methods perfect for pain detection, use of analgesics, analgesics and tranquilizers, use of radiography (tomography) to detect tumors or organic deterioration and application anticipated euthanasia [10, 13].

If the meaning of these three R's is carefully analyzed, it will be clear that these three criteria can implie a debate in which, for some researchers (teachers) or institutions, the interpretation of these guidelines may depend on questions such as the individual morality of the experimenter and the resource account for compliance with the three Rs, or even that the price

Despite these apparent obstacles, it must be clear that first individually, then institutionally and at last nationally, it is possible to gather agreements that legislate and delimitate the animal handling ethics (bioethics) following the established legal guidelines (even if there are few of them) and to try and fulfill the three Rs and the different legal regulations for the case. In some instances, attempts have been made to solve some of the suggestions of the three Rs, arguing that the required specifications would represent a considerable cost for the institution and that the budget for the experiments would increase considerably, even while using stray dogs and stray cats. The latter, of course, in the long-term could represent an additional expense when designing an experiment and generate the respective results because by not knowing the previous state of health and metabolic integrity of these animals, there would be the risk that many of them die in the course of experimental manipulation or that valid or uniform results are not obtained due to the individual variability (heterogeneity) of the animals used [4, 15]. Therefore, one would necessarily have to return to the similar approach to

In reference to Mexico, in the last two decades, laws or regulations for animal handling have been promulgated, taking into consideration the ethics aspect. For instance, on June 28, 2001, the Agriculture Ministry, Livestock, Rural Development and Fisheries and Food published in the Official Journal of the Federation the Official Mexican Standard NOM-062-ZOO-1999 "Technical specifications for the production, care and handling of laboratory animals," which specifies, among other things, "it is SAGARPA duty to promote the production, care and handling of laboratory animals through the application of techniques that assure the production,

protection health and the advantage of using laboratory animals" [14].

other more modern techniques.

that proposed by the three Rs.

avoid repetition); and specialized literature access.

increases to implement *in vivo* animal-replacement techniques.

In 1959, Russell and Burch marked a milestone in ethics in the handling of laboratory animals when they published their book titled *The Principles of Humane Experimental Technique* [13], which, through time, became a reference for animal handling. The essence of this proposal is summarized in the famous three R's:

**Figure 1.** Critical factors for the welfare of the laboratory animal.

Replacement: This includes conscious animals for unconscious animals or insensitive materials. To get this it is necessary to consider the use of *in vitro* systems, audiovisual aids, sacrificed animals, slaughterhouse material, use of invertebrates, human material, human volunteers or other more modern techniques.

As the reader will understand, the concepts mentioned above establish some criteria of laboratory animal handling, but it does not classify those considering ethical aspects about the use of species in laboratory. For this reason, in many cases, the ethical aspects about the inclusion of animal experimentation depend on the exclusive experimenter judgment, who designs the treatment to obtain results in a short or medium term to use them for human treatment or, at least, to publish the researcher's scientific results in a scientific magazine which will be rewarded with "points" for the curriculum vitae or for receiving scholarships. This leads to keep ethical aspects aside at the point of making an experiment design. This leads to putting aside the ethical aspects when designing an experiment, to which is added that in countries like Mexico, in many of the institutions where research is done, just a few years ago, it has begun to integrate committees of bioethics that evaluate and ensure that animals are treated in accordance with international ethical statutes. On the other hand, problems can arise when submitting an article to be published in a scientific journal, when an author comes from a country where there are different ethical laws that do not follow these rules to the letter [12]. Even worse, in many countries, there is an infinity of laws for the ethical animal handling regulation and the author (with the editors) must decide which one of these to follow to write the article. As an example, in the United States, in 2004, 2100 laws for animal wellbeing were proposed [8]. However, in a certain way, in 2006, it was intended to be solved when the International Committee of Medical Journal Editors recommended the authors to report if animal experiments had been conducted according to the institutional and national guidelines about the use and care of laboratory animals [12]. In general, a laboratory animal should be kept in appropriate conditions, including its storage place, its feeding and its

In 1959, Russell and Burch marked a milestone in ethics in the handling of laboratory animals when they published their book titled *The Principles of Humane Experimental Technique* [13], which, through time, became a reference for animal handling. The essence of this proposal is

genetic characteristics (**Figure 1**).

142 Reflections on Bioethics

summarized in the famous three R's:

**Figure 1.** Critical factors for the welfare of the laboratory animal.

Reduction: This includes reducing the number of animals without accuracy diminution. For this, it should consider the use of genetically homogeneous colonies without environmental influences; the animal model selected; sanitary, genetics and environmental quality; cryopreservation; advanced biostatistics methodology; data bank (publishing negative results to avoid repetition); and specialized literature access.

Refinancing: It involves pain and discomfort reduction techniques taking into consideration animal care and well-being, dexterity and training of laboratory personnel to make methods perfect for pain detection, use of analgesics, analgesics and tranquilizers, use of radiography (tomography) to detect tumors or organic deterioration and application anticipated euthanasia [10, 13].

If the meaning of these three R's is carefully analyzed, it will be clear that these three criteria can implie a debate in which, for some researchers (teachers) or institutions, the interpretation of these guidelines may depend on questions such as the individual morality of the experimenter and the resource account for compliance with the three Rs, or even that the price increases to implement *in vivo* animal-replacement techniques.

Despite these apparent obstacles, it must be clear that first individually, then institutionally and at last nationally, it is possible to gather agreements that legislate and delimitate the animal handling ethics (bioethics) following the established legal guidelines (even if there are few of them) and to try and fulfill the three Rs and the different legal regulations for the case. In some instances, attempts have been made to solve some of the suggestions of the three Rs, arguing that the required specifications would represent a considerable cost for the institution and that the budget for the experiments would increase considerably, even while using stray dogs and stray cats. The latter, of course, in the long-term could represent an additional expense when designing an experiment and generate the respective results because by not knowing the previous state of health and metabolic integrity of these animals, there would be the risk that many of them die in the course of experimental manipulation or that valid or uniform results are not obtained due to the individual variability (heterogeneity) of the animals used [4, 15]. Therefore, one would necessarily have to return to the similar approach to that proposed by the three Rs.

In reference to Mexico, in the last two decades, laws or regulations for animal handling have been promulgated, taking into consideration the ethics aspect. For instance, on June 28, 2001, the Agriculture Ministry, Livestock, Rural Development and Fisheries and Food published in the Official Journal of the Federation the Official Mexican Standard NOM-062-ZOO-1999 "Technical specifications for the production, care and handling of laboratory animals," which specifies, among other things, "it is SAGARPA duty to promote the production, care and handling of laboratory animals through the application of techniques that assure the production, protection health and the advantage of using laboratory animals" [14].

Currently, the lack of planning in the production of laboratory animals, the lack of homogeneous criteria related to the activities addressed to care, handling and utility of animals with scientific, technological development and innovation purposes have caused that the care, the treatment and the application of experimental techniques practiced in these animals be exercised in an inadequate way and, therefore, representing severe damage to animal welfare. To achieve reliable results in scientific research, biomedical teaching and quality control, as well as to minimize the number of animals available, it is necessary to have laboratory animals in optimal conditions [4].

**4. Status of bioethics in superior schools of Mexico**

cedures and along its captivity previous to laboratory handling" [23].

One more example is the National School of Biological Sciences (ENCB by its initials in Spanish) of the National Polytechnic Institute, in which was recently established, in 2008, the bioethics committee, which has issued a regulation that is periodically revised in accordance with the scientific and social changes that come through the country. This committee's achievement is that many researchers take its advice about the ethical procedures for the

These actions are trying to be implemented also in higher educational institutions in Mexico, although with some discussions and problems. Sometimes the bioethics committees of each of these educational institutions could consider that the teachers-researchers are not properly handling animals in their charge, due to the high number of them that are used both for teaching and for research. However, the researcher could argue that, especially in the biomedical area, the use of many animals distributed in several lots that are submitted to different treatments could guarantee, statistically, reliable results and results compatible with human studies. In this sense, the discussion arises when some of the bioethics local committees propose the total or partial animal replacement in this kind of experiment (or in laboratory practices for teaching courses), arguing that current technology already offers tools to elaborate computation models to simulate and even predict some effects on organisms when most of the variabilities are controlled in an experiment. As an evidence of this, some medicine schools have replaced the use of animals in classes (like pharmacology, pathology, neuroanatomy, etc.) for computer simulation models or computerized mannequins which are programmed to respond to different variables that simulate some metabolic disorders, psychological disorders and so on. However, these teaching-research methods could have the disadvantage that the student (future researcher) does not deal with real situations, where it is not enough to have the theoretical knowledge about the kind of response an animal or a human being could have when any of them are exposed to a specific experimental handling; the fact that, with those methods it maybe not cause suffering to the animals which can be considered an advantage, but the student would lose the ability to react and make decisions when handling real situations with humans and animals. Even then, it is necessary to highlight that despite these ethic-philosophic issues, many universities around the country are trying to create their own bioethics committees that work following the national and international guidelines, without removing the student training aspect that the experimental animal handling provides. As an evidence of this, it is possible to mention some institutions such as the Autonomous Juarez University in Tabasco (2010), which has published a manual for the handling of animals with experimentation and teaching purposes and in the introduction, comments that: "When it did not count on alternative tools for the use of animals and required the use of it, the procedures performed must follow a scientific and teaching justified propose, have a reasonable expectation as far as an increase of knowledge is concerned about the biological processes and provide the necessary ability for the correct technique handling. It is necessary to take account that this technique it is justified only in the case of science knowledge for the good of humanity or animals." This manual ends, arguing that: "It is obligation of who is handling animals with study purposes to provide them with a real treat and proper care, from its capture pro-

Bioethics in the Use of Experimental Animals http://dx.doi.org/10.5772/intechopen.74520 145

The above is more relevant when the experimental designs include non-human primates (NHP), due to the closeness and similarity with the human being [15]. For this reason, a workshop was organized in 2014 on "alternative methods for the use of NHP in biomedical research," under the international exchange program of European Primate Network (EUPRIM-Net II) [16], which reinforced the application of the concept of the three Rs in the improvement of the techniques for the use of non-human primates in biomedical experiments that serve for research or for education.

In other countries, such as those in the European continent, efforts are being made to improve the conditions of animals when they are used in laboratory experiments. For this, there are studies such as the "EXEMPLAR" scale, whose meaning is "Excellence in Editorial Mandatory Policies for Animal Research" [17]. This article was published in Portugal and conducted a sampling of 170 journals from 20 countries, dedicated to the dissemination of studies with animals in the medical-biological area. These studies were classified into four categories according to the publication policies used in those studies. The categories evaluated were (a) regulatory compliance, (b) quality of research and reporting of results, (c) animal welfare and ethics and (d) criteria for exclusion of papers. Although this study emphasized the good application of policies to approve a paper, describing experiments with animals, it is also made clear that there is very little progress in the policies of each publisher about the ethical treatment of animals [17]. This may be because many researchers still do not recognize that laboratory animals are vulnerable living beings to which they must be recognized as a great part of the raw material for the advancement of science [18].

At this point it is good to comment on a study carried out with 217 students from two university faculties in Silesia (Poland) in the year 2015, who were asked to answer a questionnaire on issues such as "to granting animals personality, consciousness and the right to life," and although no differences were found in their responses due to gender, religion, educational level and so on, it was seen that they had very little knowledge of animal protection laws and about alternative methods in animal research [19].

In 2016, another work was published based on previous reviews on the freedoms that should be granted to laboratory animals to maintain their well-being [20, 21], and five provisions were proposed to ensure non-abuse of laboratory animals, such as (1) good nutrition, (2) good environment, (3) good health, (4) appropriate behavior and (5) positive mental experiences—all this in order to make a clear guide for the management of animals, both for researchers and for people not specialized in the subject but who work in research laboratories [22].

## **4. Status of bioethics in superior schools of Mexico**

Currently, the lack of planning in the production of laboratory animals, the lack of homogeneous criteria related to the activities addressed to care, handling and utility of animals with scientific, technological development and innovation purposes have caused that the care, the treatment and the application of experimental techniques practiced in these animals be exercised in an inadequate way and, therefore, representing severe damage to animal welfare. To achieve reliable results in scientific research, biomedical teaching and quality control, as well as to minimize the number of animals available, it is necessary to have laboratory animals in

The above is more relevant when the experimental designs include non-human primates (NHP), due to the closeness and similarity with the human being [15]. For this reason, a workshop was organized in 2014 on "alternative methods for the use of NHP in biomedical research," under the international exchange program of European Primate Network (EUPRIM-Net II) [16], which reinforced the application of the concept of the three Rs in the improvement of the techniques for the use of non-human primates in biomedical experiments

In other countries, such as those in the European continent, efforts are being made to improve the conditions of animals when they are used in laboratory experiments. For this, there are studies such as the "EXEMPLAR" scale, whose meaning is "Excellence in Editorial Mandatory Policies for Animal Research" [17]. This article was published in Portugal and conducted a sampling of 170 journals from 20 countries, dedicated to the dissemination of studies with animals in the medical-biological area. These studies were classified into four categories according to the publication policies used in those studies. The categories evaluated were (a) regulatory compliance, (b) quality of research and reporting of results, (c) animal welfare and ethics and (d) criteria for exclusion of papers. Although this study emphasized the good application of policies to approve a paper, describing experiments with animals, it is also made clear that there is very little progress in the policies of each publisher about the ethical treatment of animals [17]. This may be because many researchers still do not recognize that laboratory animals are vulnerable living beings to which they must be recognized as a great

At this point it is good to comment on a study carried out with 217 students from two university faculties in Silesia (Poland) in the year 2015, who were asked to answer a questionnaire on issues such as "to granting animals personality, consciousness and the right to life," and although no differences were found in their responses due to gender, religion, educational level and so on, it was seen that they had very little knowledge of animal protection laws and

In 2016, another work was published based on previous reviews on the freedoms that should be granted to laboratory animals to maintain their well-being [20, 21], and five provisions were proposed to ensure non-abuse of laboratory animals, such as (1) good nutrition, (2) good environment, (3) good health, (4) appropriate behavior and (5) positive mental experiences—all this in order to make a clear guide for the management of animals, both for researchers and for people not specialized in the subject but who work in research labo-

optimal conditions [4].

144 Reflections on Bioethics

that serve for research or for education.

part of the raw material for the advancement of science [18].

about alternative methods in animal research [19].

ratories [22].

These actions are trying to be implemented also in higher educational institutions in Mexico, although with some discussions and problems. Sometimes the bioethics committees of each of these educational institutions could consider that the teachers-researchers are not properly handling animals in their charge, due to the high number of them that are used both for teaching and for research. However, the researcher could argue that, especially in the biomedical area, the use of many animals distributed in several lots that are submitted to different treatments could guarantee, statistically, reliable results and results compatible with human studies. In this sense, the discussion arises when some of the bioethics local committees propose the total or partial animal replacement in this kind of experiment (or in laboratory practices for teaching courses), arguing that current technology already offers tools to elaborate computation models to simulate and even predict some effects on organisms when most of the variabilities are controlled in an experiment. As an evidence of this, some medicine schools have replaced the use of animals in classes (like pharmacology, pathology, neuroanatomy, etc.) for computer simulation models or computerized mannequins which are programmed to respond to different variables that simulate some metabolic disorders, psychological disorders and so on. However, these teaching-research methods could have the disadvantage that the student (future researcher) does not deal with real situations, where it is not enough to have the theoretical knowledge about the kind of response an animal or a human being could have when any of them are exposed to a specific experimental handling; the fact that, with those methods it maybe not cause suffering to the animals which can be considered an advantage, but the student would lose the ability to react and make decisions when handling real situations with humans and animals. Even then, it is necessary to highlight that despite these ethic-philosophic issues, many universities around the country are trying to create their own bioethics committees that work following the national and international guidelines, without removing the student training aspect that the experimental animal handling provides. As an evidence of this, it is possible to mention some institutions such as the Autonomous Juarez University in Tabasco (2010), which has published a manual for the handling of animals with experimentation and teaching purposes and in the introduction, comments that: "When it did not count on alternative tools for the use of animals and required the use of it, the procedures performed must follow a scientific and teaching justified propose, have a reasonable expectation as far as an increase of knowledge is concerned about the biological processes and provide the necessary ability for the correct technique handling. It is necessary to take account that this technique it is justified only in the case of science knowledge for the good of humanity or animals." This manual ends, arguing that: "It is obligation of who is handling animals with study purposes to provide them with a real treat and proper care, from its capture procedures and along its captivity previous to laboratory handling" [23].

One more example is the National School of Biological Sciences (ENCB by its initials in Spanish) of the National Polytechnic Institute, in which was recently established, in 2008, the bioethics committee, which has issued a regulation that is periodically revised in accordance with the scientific and social changes that come through the country. This committee's achievement is that many researchers take its advice about the ethical procedures for the medical-biological sciences experimentation, and for the teaching aspect, it has seen to it that the practice manuals of subjects such as human physiology, general physiology and pharmacology systems include instructions for the animal handling and slaughter in accordance with the corresponding standards, and in the case of human experimentation (students), a questionnaire is filled and signed by parents or legal tutors specifying the type of experimentation they are participating in. In the mentioned regulation, the bioethics committee of the ENCB propose as a principal objective the following: "To establish and to enforce the fundamental ethics principles in the human experimentation, and to assure a minimal suffering to animal handling in laboratory" [24]. It should be necessary to keep a balance between institutionteacher and researcher-user for the development of the medical, biological and technological knowledge, focusing on society and the own subject of investigation so that the established goals might be reached.

elementary school so that when they enter a higher-level school, they have the principles of

Bioethics in the Use of Experimental Animals http://dx.doi.org/10.5772/intechopen.74520 147

It is the task of all of us who are dedicated to scientific work to be aware of the provisions that will surely apply in the future regarding the ethical management of animals, all this always in

The authors want to thank the ENCB-IPN for the support received for this work through the

Departamento de Fisiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico

[1] Sechzer JA. The role of the animals in biomedical research. Annals of the New York

[3] Pérez-García CC, Diez-Prieto MI, García-Partida P. Introducción a la experimentación y

[4] De Aluja AS. Animales de laboratorio y la Norma Oficial Mexicana (NOM-062-

[5] Mrad-De-Osorio A. Ética en la investigación con modelos animales experimentales. Alternativas y las 3 RS de Russel. Una responsabilidad y un compromiso ético que nos

[2] Dolan K. Ethics, Animals and Science. London: Blackwell Science Ltd; 1999. 210 p

protección animal. León España : Universidad de León; 1999. 150 p

compete a todos. Revista Colombiana de Bioética. 2006;**1**(1):163-183

ZOO-1999). Gaceta Médica de México. 2002;**138**(3):295-298

animal welfare in scientific research as a basic principle of their academic training.

The authors have no conflict of interest in the publishing of this material.

Tomás Alejandro Fregoso Aguilar\* and Elizabeth Guarneros Bañuelos

\*Address all correspondence to: fisiobiologo@hotmail.com

Academy of Sciences New York. 1983;**406**:1-229

the constant search for knowledge.

**Acknowledgements**

SIP Project 20170800.

**Conflict of interest**

**Author details**

**References**

Nacional, México City, Mexico

The functionality and authority of this committee have been developed in all this time so that the regulations and the established rules are complied with and respected under a legal framework. Of course it is important to take into consideration the National Autonomous University of Mexico' efforts, which has bioethics committees focusing on the same principles' optimization in the use of animals avoiding senseless suffering. As an evidence of this rationale, it is possible to mention the medicine faculty publication (Research coordination, ethic committee) titled *Ethical considerations for the usage of experimentation with animals in research projects* [25], in which detailed specifications of the type of facilities about animal accommodation are listed and what they should consider to provide animals shelter, the appropriate equipment, feeding issues, water provision as well as experimental techniques that include analgesia, anesthetic and euthanasia according to the regulation NOM-062-ZOO-1999.

## **5. Conclusion**

From the abovementioned, it is evident and obvious that, although it has achieved a great deal of progress in the ethical field toward experimentation animal handling, there are still agreements to reach, based on legal, moral and ethical procedures that allow respect for all those species used in experimentation and, at the same time, obtain reliable experimentation results to justify its implementation in science (and teaching).

It would be desirable to achieve uniform acceptance of the concepts of animal bioethics already in use in some countries of the American continent, with the most recent proposals arising in Europe. Perhaps this could be achieved by combining the concepts outlined in the three Rs with the proposal of the five provisions for the welfare of a laboratory animal. In addition, this must, perhaps, be reflected in international bioethics laws that not only establishes the guidelines followed for a good handling of laboratory animals but also to impose legal sanctions for those investigators or institutions that inflict harm to animals.

Finally, we must bear in mind that knowing how to manipulate a laboratory animal implies the education of the researcher, so students should be educated in these aspects from the elementary school so that when they enter a higher-level school, they have the principles of animal welfare in scientific research as a basic principle of their academic training.

It is the task of all of us who are dedicated to scientific work to be aware of the provisions that will surely apply in the future regarding the ethical management of animals, all this always in the constant search for knowledge.

## **Acknowledgements**

medical-biological sciences experimentation, and for the teaching aspect, it has seen to it that the practice manuals of subjects such as human physiology, general physiology and pharmacology systems include instructions for the animal handling and slaughter in accordance with the corresponding standards, and in the case of human experimentation (students), a questionnaire is filled and signed by parents or legal tutors specifying the type of experimentation they are participating in. In the mentioned regulation, the bioethics committee of the ENCB propose as a principal objective the following: "To establish and to enforce the fundamental ethics principles in the human experimentation, and to assure a minimal suffering to animal handling in laboratory" [24]. It should be necessary to keep a balance between institutionteacher and researcher-user for the development of the medical, biological and technological knowledge, focusing on society and the own subject of investigation so that the established

The functionality and authority of this committee have been developed in all this time so that the regulations and the established rules are complied with and respected under a legal framework. Of course it is important to take into consideration the National Autonomous University of Mexico' efforts, which has bioethics committees focusing on the same principles' optimization in the use of animals avoiding senseless suffering. As an evidence of this rationale, it is possible to mention the medicine faculty publication (Research coordination, ethic committee) titled *Ethical considerations for the usage of experimentation with animals in research projects* [25], in which detailed specifications of the type of facilities about animal accommodation are listed and what they should consider to provide animals shelter, the appropriate equipment, feeding issues, water provision as well as experimental techniques that include analgesia, anesthetic and euthanasia according to the regulation NOM-062-ZOO-1999.

From the abovementioned, it is evident and obvious that, although it has achieved a great deal of progress in the ethical field toward experimentation animal handling, there are still agreements to reach, based on legal, moral and ethical procedures that allow respect for all those species used in experimentation and, at the same time, obtain reliable experimentation

It would be desirable to achieve uniform acceptance of the concepts of animal bioethics already in use in some countries of the American continent, with the most recent proposals arising in Europe. Perhaps this could be achieved by combining the concepts outlined in the three Rs with the proposal of the five provisions for the welfare of a laboratory animal. In addition, this must, perhaps, be reflected in international bioethics laws that not only establishes the guidelines followed for a good handling of laboratory animals but also to impose

Finally, we must bear in mind that knowing how to manipulate a laboratory animal implies the education of the researcher, so students should be educated in these aspects from the

legal sanctions for those investigators or institutions that inflict harm to animals.

results to justify its implementation in science (and teaching).

goals might be reached.

146 Reflections on Bioethics

**5. Conclusion**

The authors want to thank the ENCB-IPN for the support received for this work through the SIP Project 20170800.

## **Conflict of interest**

The authors have no conflict of interest in the publishing of this material.

## **Author details**

Tomás Alejandro Fregoso Aguilar\* and Elizabeth Guarneros Bañuelos

\*Address all correspondence to: fisiobiologo@hotmail.com

Departamento de Fisiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, México City, Mexico

## **References**


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[22] Mellor DJ. Moving beyond the "five freedoms" by updating the "five provisions" and

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[23] Falconi E, García ML, Marín RO, Padrón LRM, Rivas AMG, Vargas SG. Manual para el manejo de animales con fines de experimentación y enseñanza. México: Universidad

[24] ENCB. Reglamento del comité de bioética. Escuela Nacional de Ciencias Biológicas.

[25] Facultad de medicina de la UNAM. Consideraciones éticas para el caso de utilización de animales de experimentación en los proyectos de investigación. Comisión de ética de la

introducing aligned "animal welfare aims". Animals. 2016;**6**(59):1-7

Juárez Autónoma de Tabasco; 2010. p. 14

Fac. de Medicina. UNAM. México. 2007. p. 13

México: IPN; 2008. p. 15


[22] Mellor DJ. Moving beyond the "five freedoms" by updating the "five provisions" and introducing aligned "animal welfare aims". Animals. 2016;**6**(59):1-7

[6] Singer P. The Animal Liberation Movement. 1985. Available from: www.nwveg.org/

[7] UNESCO. Universal Declaration of Animal Rights. 1978. Available from: http://con-

[8] Rollin BE. Ethics and euthanasia. The Canadian Veterinary Journal. 2009;**50**:1081-1086 [9] Zuñiga JM, Tur JA, Milocco SN, Piñeiro R. Ciencia y tecnología en protección y experi-

[10] Mrad-De-Osorio A, Rosenkranz A. Guía para el uso de animales de laboratorio, parte

[11] Tobón-Marulanda FA. Principios básicos de la farmacología experimental. Medellín,

[12] Clutton RE. Clinical studies, pain and ethics. The Journal of Small Animal Practice.

[13] Russell WMS, Burch RL. The Principles of Humane Experimental Technique. London, UK: Methuen; 1959. 238 p. [Special edition published in 1992 by Universities Federation

[14] NOM-062-ZOO-1999. Especificaciones técnicas para la producción, cuidado y uso de los animales de laboratorio. Available from: www.fmvz.unam.mx/fmvz/principal/

[15] Zhang XL, Pang W, Hu XT, Li JL, Yao YG, Zheng YT. Experimental primate and nonhuman primate (NHP) models of human diseases in China: Current status and progress.

[16] Burm SM, Prins JB, Langermans J, Bajramovic J. Alternative methods for the use of non-human primates in biomedical research. ALTEX Journal. 31 2014. http://dx.doi.

[17] Martins AR, Franco NH. A critical look at biomedical journals' policies on animal research by use of a novel tool: The EXEMPLAR scale. Animals. 2015;**5**:315-331

[18] Johnson J. Vulnerable subjects? The case of nonhuman animals in experimentation.

[19] Fedyk O, Wilczynski KM. Influence of selected factors on development the student's of the biological sciences faculties bioethical attitudes towards experimental animals stud-

[20] Webster J. Assessment of Animal Welfare: The five Freedoms. In Animal Care: A Cool

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I. Bogotá, Colombia: Universidad Nacional de Colombia; 1990. 147 p


**Chapter 9**

**Provisional chapter**

**Alternatives to Animal Experimentation: Its**

**Alternatives to Animal Experimentation: Its** 

DOI: 10.5772/intechopen.74941

Although it is desirable to replace scientific procedures with live animals by other methods that do not use them, the use of animals in scientific procedures should be restricted to those areas that benefit human, animal, and environmental health. The use of animals as experimental models of observation of biological phenomena has evolved with man, to this day. The use of animals for scientific or educational purposes should be considered only when there is no other alternative and it is governed by the principles of replacement, reduction, and refinement. The scientists should be sure that the information obtainable with the experiments is not yet available or that the protocol was designed taking into account animal protection considerations. The chosen methods must use the least number of animals; provide satisfactory results; use the species with the least ability to experience pain, suffering, anguish, and damage; and be optimal for the extrapolation of results to the target species such as humans. It will be fundamental to guarantee on a scientific and ethical basis that the use of an animal is subject to a careful evaluation regarding the scientific or educational validity. **Keywords:** animal experimentation, animal model, laboratory animals, research design,

The research is focused today on the ethical, logistical, economic, scientific, and legal requirements. At the European level, Directive 2010/63 of the European Parliament and of

> © 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

© 2018 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use,

distribution, and reproduction in any medium, provided the original work is properly cited.

**Institutional Teaching and Scientific**

**Institutional Teaching and Scientific**

Additional information is available at the end of the chapter

Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/intechopen.74941

animal testing alternatives, animal

**Abstract**

**1. Introduction**

Liliana Anguiano-Robledo, Claudia C. Calzada-Mendoza, Cornelio Barrientos-Alvarado and María Elena Hernández-Campos

Liliana Anguiano-Robledo, Claudia C. Calzada-Mendoza, Cornelio Barrientos-Alvarado and María Elena Hernández-Campos

#### **Chapter 9 Provisional chapter**

#### **Alternatives to Animal Experimentation: Its Institutional Teaching and Scientific Alternatives to Animal Experimentation: Its Institutional Teaching and Scientific**

DOI: 10.5772/intechopen.74941

Liliana Anguiano-Robledo, Claudia C. Calzada-Mendoza, Cornelio Barrientos-Alvarado and María Elena Hernández-Campos Liliana Anguiano-Robledo, Claudia C. Calzada-Mendoza, Cornelio Barrientos-Alvarado and María Elena Hernández-Campos

Additional information is available at the end of the chapter Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/intechopen.74941

#### **Abstract**

Although it is desirable to replace scientific procedures with live animals by other methods that do not use them, the use of animals in scientific procedures should be restricted to those areas that benefit human, animal, and environmental health. The use of animals as experimental models of observation of biological phenomena has evolved with man, to this day. The use of animals for scientific or educational purposes should be considered only when there is no other alternative and it is governed by the principles of replacement, reduction, and refinement. The scientists should be sure that the information obtainable with the experiments is not yet available or that the protocol was designed taking into account animal protection considerations. The chosen methods must use the least number of animals; provide satisfactory results; use the species with the least ability to experience pain, suffering, anguish, and damage; and be optimal for the extrapolation of results to the target species such as humans. It will be fundamental to guarantee on a scientific and ethical basis that the use of an animal is subject to a careful evaluation regarding the scientific or educational validity.

**Keywords:** animal experimentation, animal model, laboratory animals, research design, animal testing alternatives, animal

#### **1. Introduction**

The research is focused today on the ethical, logistical, economic, scientific, and legal requirements. At the European level, Directive 2010/63 of the European Parliament and of

© 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. © 2018 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

the Council of September 22, 2010, on the protection of animals used for scientific purposes [1] must be highlighted, which is translated into Spanish legislation by Royal Decree 53/2013 [1, 2]. Researchers have to demonstrate the real need to use animals in scientific and teaching applications. These regulations aim to ensure animal protection and, in particular, adequate care for animals; not unnecessarily cause pain, suffering, anguish or prolonged injury; avoid duplication of procedures; minimize the number of animals used in procedures; and apply possible alternative methods.

**4.** Evaluation: detection, regulation, or modification of physiological conditions in humans,

Alternatives to Animal Experimentation: Its Institutional Teaching and Scientific

http://dx.doi.org/10.5772/intechopen.74941

153

**5.** The welfare of animals, particularly the improvement of the conditions of production of

**8.** Protection of the natural environment in favor of the welfare of human beings or animals

The use of animals in scientific experiments likely to cause pain, distress, or lasting harm generates important ethical issues. Animals should be used only if the scientific objectives are valid, there is no other alternative, and the cost to the animals is not excessive. "Validity" in this case implies that the experiment has a high probability of meeting the stated objectives, and these objectives have a reasonable chance of contributing to human or animal welfare,

Scientists who use animals in research must justify the number of animals to be used, and committees that review proposals to use animals must review this justification to ensure the appropriateness of the number of animals to be used. Obtaining satisfactory scientific results, it will depends of sample size calculation should be performed as well as, the election of more

Regulation D2010/63/EU aims to establish the applicable standards for the protection of animals used, for scientific purposes [6]. It establishes for the first time in EU legislation the principle of "the three Rs" and imposes it as a firm legal requirement in all aspects of the care and use of animals in that area. The directive, in its application, goes beyond the initial interpretation and also regulates the breeding and care of the animals, that is, guarantees refinement during housing, breeding, and care, even if the animal is not object of any scientific procedure,

**2.** The use of animals for scientific or teaching purposes should be considered only when

**4.** Ethical considerations in the use of animals are the basis of the authorization of projects.

**1.** The experimental protocol should be with respect to the project objectives.

**3.** The objectives cannot be achieved by alternative methods.

**9.** Higher education or training for the acquisition or improvement of professional skills

**6.** Evaluate the efficacy and safety of new pharmaceutical products

**7.** Research directed to the conservation of the species

**2. Criteria for the evaluation of a project**

animals, or plants

**10.** Legal and forensic medicine

possibly in the long term [4].

suitable animals [5].

regulating the following:

there is no alternative.

animals

Russell and Burch [3] formulated for the first time the "principle of the three Rs" that was adopted by the aforementioned regulations. Russell and Burch considered that the replacement was the ultimate goal of the investigation. Its main message is, in summary, that, if we are to use a criterion to choose which experiments to carry out, that of humanity is the best we can ever conceive and that the greatest scientific achievements have always been the most human and the most esthetically attractive, those that best transmit that sense of beauty and elegance that constitutes the very essence of science in its best aspect. Animals should be replaced by less sentient alternatives such as invertebrates or *in vitro* methods whenever possible. Only an experiment with live animals should be carried out if there is no alternative method for the procedure we wish to perform (replace), for example, using audiovisual media or virtual reality techniques [4].

Secondly, if the alternative method does not exist and we have to perform the experiment with live animals, the number of animals should be reduced to the minimum consistent with the scientific objectives of the study, recognizing that important biological effects may be missed if too few animals are used [4].

And thirdly, we are also told that we must modify the procedures used so that animals suffer as little as possible (refine). Experimental protocols should be refined to minimize any adverse effects for each individual animal. For example, appropriate anesthesia and analgesia should be used for any surgical intervention. Death is not an acceptable endpoint if it is preceded by some hours of acute distress, and humane endpoints should be used whenever possible. Staff should be well trained, and housing should be of a high standard with appropriate environmental enrichment. Animals should be protected from pathogens [4].

Its main message is, in summary, that, if we are to use a criterion to choose which experiments to carry out, that of humanity is the best we can ever conceive and that the greatest scientific achievements have always been the most human and the most esthetically attractive, those that best transmit that sense of beauty and elegance that constitutes the very essence of science in its best aspect. Royal Decree 53/2013 aims to establish the applicable standards for the protection of animals used, bred, or supplied for the purpose of experimentation and other scientific purposes, including education and teaching. For this, it regulates the following:


the Council of September 22, 2010, on the protection of animals used for scientific purposes [1] must be highlighted, which is translated into Spanish legislation by Royal Decree 53/2013 [1, 2]. Researchers have to demonstrate the real need to use animals in scientific and teaching applications. These regulations aim to ensure animal protection and, in particular, adequate care for animals; not unnecessarily cause pain, suffering, anguish or prolonged injury; avoid duplication of procedures; minimize the number of animals used in procedures; and apply

Russell and Burch [3] formulated for the first time the "principle of the three Rs" that was adopted by the aforementioned regulations. Russell and Burch considered that the replacement was the ultimate goal of the investigation. Its main message is, in summary, that, if we are to use a criterion to choose which experiments to carry out, that of humanity is the best we can ever conceive and that the greatest scientific achievements have always been the most human and the most esthetically attractive, those that best transmit that sense of beauty and elegance that constitutes the very essence of science in its best aspect. Animals should be replaced by less sentient alternatives such as invertebrates or *in vitro* methods whenever possible. Only an experiment with live animals should be carried out if there is no alternative method for the procedure we wish to perform (replace), for example, using audiovisual media or virtual reality techniques [4]. Secondly, if the alternative method does not exist and we have to perform the experiment with live animals, the number of animals should be reduced to the minimum consistent with the scientific objectives of the study, recognizing that important biological effects may be

And thirdly, we are also told that we must modify the procedures used so that animals suffer as little as possible (refine). Experimental protocols should be refined to minimize any adverse effects for each individual animal. For example, appropriate anesthesia and analgesia should be used for any surgical intervention. Death is not an acceptable endpoint if it is preceded by some hours of acute distress, and humane endpoints should be used whenever possible. Staff should be well trained, and housing should be of a high standard with appropriate environ-

Its main message is, in summary, that, if we are to use a criterion to choose which experiments to carry out, that of humanity is the best we can ever conceive and that the greatest scientific achievements have always been the most human and the most esthetically attractive, those that best transmit that sense of beauty and elegance that constitutes the very essence of science in its best aspect. Royal Decree 53/2013 aims to establish the applicable standards for the protection of animals used, bred, or supplied for the purpose of experimentation and other scientific purposes, including education and teaching. For this, it regulates the following:

**2.** The application of the scientific method in which a problem is first identified and observa-

**3.** Prevention: prophylaxis, diagnosis or treatment of diseases, or their effects on humans,

mental enrichment. Animals should be protected from pathogens [4].

tions, or other relevant data are then used to construct a solution:

possible alternative methods.

152 Reflections on Bioethics

missed if too few animals are used [4].

**1.** Basic investigation

animals, or plants

The use of animals in scientific experiments likely to cause pain, distress, or lasting harm generates important ethical issues. Animals should be used only if the scientific objectives are valid, there is no other alternative, and the cost to the animals is not excessive. "Validity" in this case implies that the experiment has a high probability of meeting the stated objectives, and these objectives have a reasonable chance of contributing to human or animal welfare, possibly in the long term [4].

Scientists who use animals in research must justify the number of animals to be used, and committees that review proposals to use animals must review this justification to ensure the appropriateness of the number of animals to be used. Obtaining satisfactory scientific results, it will depends of sample size calculation should be performed as well as, the election of more suitable animals [5].

## **2. Criteria for the evaluation of a project**

Regulation D2010/63/EU aims to establish the applicable standards for the protection of animals used, for scientific purposes [6]. It establishes for the first time in EU legislation the principle of "the three Rs" and imposes it as a firm legal requirement in all aspects of the care and use of animals in that area. The directive, in its application, goes beyond the initial interpretation and also regulates the breeding and care of the animals, that is, guarantees refinement during housing, breeding, and care, even if the animal is not object of any scientific procedure, regulating the following:


**5.** The application of the principles of replacement, reduction, and refinement must be guaranteed.

**3. Importance of animal experimentation**

able data than outbred stocks [11].

**4. Choice of animal model**

model (negative controls) [12].

**4.1. Classification of animal models**

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

Alternatives to Animal Experimentation: Its Institutional Teaching and Scientific

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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 repeat-

One of the uses of animal models is related to the evaluation of new drugs for the treatment of human diseases. For this type of use, the animal model must respond adequately to the effects of different therapeutic agents. The failure rate of investigative new drugs is excessively high, ranging from about 80 to 97% depending on the therapeutic area. 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. 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

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:


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. 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. These methods used in animal studies are explained in [8]. Systematic reviews "can help improve the methodological quality of animal experiments, make the choice of an animal model and the translation of animal data to the clinic more evidence-based and implement the 3Rs," according to [9].


**Table 1.** Considerations in the study design.
