Progress in animal experimentation ethics . A case study from a brazilian medical school and from the international medical literature

PURPOSE: This study describes in Brazil and in the global biomedical community the time course of the development of animal research welfare guidelines. METHODS: The database of the Ethics Committee of the Faculty of Medicine of Ribeirao Preto (EC/FMRP-USP), Brazil, was surveyed since its inception in 2002 as the regulations became more stringent to provide better protection of animal research welfare at this institution. Medline database was evaluated to identify the number of publications in the period between 1968 and 2008 that used research animals and were in compliance with established ethics guidelines. RESULTS: The EC/FMRP-USP evaluated 979 projects up until 2009. Most of the applications came from Department of Physiology and the most frequently requested species was the rat. In 2004, national research funding agencies started to request prior approval from institutional review ethics committees prior to application review and this requirement became federal law in Brazil in 2008. The analysis of international publications revealed a relative reduction in studies involving research animals (18% in 1968 to 7.5% in 2008). CONCLUSIONS: The present work showed that in the last four decades major changes occurred in the guidelines dictating use of research animals occurred and they are being adopted by developing countries. Moreover, animal welfare concern in the scientific community preceded the introduction of journal guidelines for this purpose. Furthermore, in Brazil it was anticipated that laws were needed to protect animal research welfare from being not upheld.


Introduction
For many centuries extending back to a time nearly twelve thousand years that predates written records when agriculture and animal domestication started, animals have been used for research by humans from different cultures. Their usage provided answers to questions pertaining to health and welfare.
This historical perspective demonstrates how effective and essential it has been to use animals as test subjects to provide in some cases life-saving answers to numerous biomedical questions.
Despite these important advances, extremist organizations have come into existence that are strongly opposed to the continued use of animals for biomedical research in the laboratory. They have launched a "Research Animal War" 1-2 whose objective is to prohibit all types of animal research.
In the nineteenth century in the United Kingdom, emerging public concern about the ethical use of animals resulted in the establishment of legal guidelines that defined procedures to be implemented protecting them from being used in an abusive manner. Such concern about the protection of animal welfare rights became important on an international scale as indicated by the passage in 1966 in the USA of the Animal Welfare Act 3 . Indeed, new animal welfare research guidelines are being continuously codified by journals, institutions and research funding agencies requiring researcher compliance before initiating experimental studies. However, the chronology and impact of developing methodological requirements on research endeavors have not been addressed before now 4-6 .
Brazil is among the twenty leading countries in the world whose number of biomedical publications is having a positive impact in different disciplines. The Faculty of Medicine at Ribeirao Preto, Sao Paulo University (FMRP-USP) is a major biomedical research center in Brazil that provides high quality contributions to biomedical research and has received attention for its potential growth due to favorable economic support [7][8][9] .
The local Animal Research Ethical Committee was created in 2002 (http://www.fmrp.usp.br/cetea) 10 , following guidelines of "Colégio Brasileiro de Experimentação Animal" (COBEA), which recently changed its name to "Sociedade Brasileira de Ciência de Animal de Laboratório", SBCal (http://www.cobea.org.br) 11 . This committee was created six years ahead of the establishment of a Brazilian Animal Welfare Research legal code. Recommendations on how to be in compliance with international guidelines were already published in Brazilian biomedical journals for more than a decade. This was already initiated when it was estimated that only 20% of the world-wide journals presented specific animal welfare research guidelines in the instructions for authors 12 .    The most used specie was rat, followed by mice, however several projects mentioned a broad range of animals, as rabbits, bovines, pigs, snakes, goats, lizard, hamsters, embryonated eggs ( Figure 3).  (Table 1). However the percentage of studies involving experimental animals decreased over 10% in 40 years. The incidence of animal usage was lower in each year except in 1998 and 2008 (Table 1).

FMRP-USP animal research Ethical Committee
The frequency of usage of different species revealed a decrease in studies involving bigger animals (e.g.; dog and pig) whereas smaller animals were used more frequently (rodents) ( Figure 4).  Journals requiring authors to sign a statement that they are in compliance with guidelines established to guarantee animal research welfare is a newly established requirement event.
Nevertheless, the proportion of journals that still have no clear policy regarding this issue is still higher than 50% 17 and in such cases, authors do not state compliance to any guidelines. It is possible that the requirement by core journals to require authors to provide assurance of adherence to animal research welfare guidelines occurred in response to institutions requiring such a guarantee from investigators prior to obtaining approval for submission of their research applications to external funding agencies. This is tenable since such an institutional requirement was established four years prior to it being instituted by core journals

Conclusions
The present study showed that in recent decades, there is an increasing concern about establishing animal research welfare guidelines by biomedical research institutions and journal publishers. Meeting this objective involves optimizing efforts by peer research task forces, funding agencies and journal publishers to scrutinize applications and manuscripts for guarantees of adherence to animal research guidelines.