Animal Research in a Global Environment: Meeting the Challenges: Proceedings of the November 2008 International Workshop (2011)

GLOBAL ISSUES: WORKING ACROSS DIFFERENT STANDARDS

Operational Challenges—Pharmaceutical Industry

Margaret Landi

My presentation is about possible solutions to different operational challenges that might exist in pharmaceutical industries, especially those that tend to work around the globe. I will cover four major topics: the challenges, the criticisms, the charge, the consensus.

The best way to start this topic is to make sure we are all working from the same dictionary, using the same definitions. As an example, the word “standard” is often used interchangeably with words like “principle” or “goal” or “objective.” A standard is something formally established as one moves forward in discussions on whatever aspect we are talking about; in this case we are talking about animal care and welfare in the pharmaceutical industry.

Sometimes we use the word “principles.” A principle is really a code of conduct. According to what principles are we going to perform studies in animals in multinational pharmaceutical companies?

The last word we often hear is “guidance.” Whether it’s the Guide for the Care and Use of Laboratory Animals, guidance, as you might suspect, deals with influencing, trying to influence a certain way, trying to achieve a particular outcome.

I will give you a perspective from my responsibility at GlaxoSmithKline (GSK), which spreads between the US and the UK and continental Europe. I also have fairly strong ties with our new R&D center in China, which, in turn, has ties with a site in Singapore where we are doing some animal research. We run the gamut from transgenic mice and transgenic rats to large nonhuman primates, macaque species.

What are the challenges when we address global guidance, how will we move forward? As you have probably heard or surmised already:

• There is a lack of consensus on what best practice is across various cultures.

• The regulations differ across countries.

• The regulations change.

• There are differences in cultural thinking.

For the last point, I would like to describe the main influences that drive differences in European versus American culture when it comes to animal research. In Europe, as opposed to the US, many things are uniform, because things are the equivalent of federally driven, whether it’s a speed limit that goes across the country, whether it’s VAT or sales tax across the country. In the US, every state is different. [For example,] the sales tax will be different—we all know the states that don’t have sales tax, and if you live near them you tend to visit them so you can get a break of 6 to 7 percent on whatever you are buying. There is a lot of diversity in the US as far as the way we expect things to happen. I’m originally from New Jersey; I travel back there a lot for family. I can use a cell phone in the state of Pennsylvania, and the minute I cross the New Jersey state border I put it away because I’m not allowed to use it unless I have a hands-free model. This is an example of differences.

Our way of living sometimes translates into the way we think about things. Generally speaking, in Europe, many things are consensus-driven—people say “let’s have a discussion.” Right now, there is the European Directive, driving similar practices in all EU countries. The US, for better or for worse, tends to work on threshold: We tolerate a lot, whether it’s national debt or handguns—whatever it happens to be—until a certain threshold is met, and then things happen. The good news is, when a threshold is met, something usually happens quickly. The bad news is that it generally takes a while to hit a threshold. The reverse would be in consensus: sometimes it takes a while to get to that outcome, but people are talking about a topic and thinking about it.

To me, the important question is, Do the variations that exist really result in a big difference in the way we care for animals in different parts of the world? Is it possible to align principles, independent of differing standards, which are the more prescriptive way of looking at animals in research?

Some of the differences, compared to academia:

• Industry usually has a very large internal capacity, there are many things we can do internally. [For example,] we generally have the option to buy equipment if we feel it’s needed and we can defend the budget for doing so.

• Industry tends to be a regulated environment. Even the earlier work has probably many more regulations than are normally seen in academia.

• Industry has many different requirements because it spans different countries and includes different types of studies, from efficacy models in transgenic animals to highly regulated GLP studies in macaques.

• Of course, timelines are extremely important.

What then is the criticism? I distilled it down to the concept that “what we think weighs more than what they think.” When we have conversations, when we have dialogues, we have already sort of made that internal critique. We need to move away from that preconceived notion before we can start to have the conversation. Otherwise, it will be hard to have consensus or even a conversation.

Again, using GSK as an example, we work across two main cultures, the US culture and the UK (we also have sites in continental Europe, but that is a more modest operation). We have opened some discussions to see if we could come to consensus on guiding principles for the use of animals in research. We did this because we felt it would provide a safe structure for having conversations and would allow the diversity of thought and opinion to come through and have a substantive type of conversation, and try to get some alignment.

Our desired outcome is to achieve alignment and articulate a set of foundation principles with regard to the animals, for us within the institution and also for work done for our institution by CROs, academic institutions, or other places. We started with dogs and monkeys, some of the more highly emotive species. We also defined what the undesired outcome would be. If we couldn’t come to some sort of understanding, that wouldn’t mean that differences in practices or standards always equate to a difference in care. We wanted to avoid a perception of a dual standard—i.e., a practice done in one place means greater care, not doing it in another place means lesser care. Unfortunately, this is often the initial perception when working in different environments.

Going back to the definitions, consensus, of course, means a general agreement and some sort of alignment or solidarity along a certain belief or sentiment. When we achieve a consensus, I think we can start to move the scales—each side will continue to maintain its own thoughts, but we will get a little more alignment and a little more agreement to ask “What is our objective?” Our objective is the care and welfare of the animals; it’s not to say that one side’s opinion is more important than the other. So let’s keep that in mind as we have the conversations.

But the question always asked is, “Is it truly achievable?” Even in societies where there are many consensus-driven processes it is sometimes difficult to achieve consensus. So my charge has been, How do we handle this?

At GSK, we have certain principles by which we work. We adhere to the 3Rs and we meet all applicable rules and regulations regarding animals in research. We have also established seven core principles of animal care and welfare. These principles are used no matter where we are in the world—whether we do research in China, North Carolina, or Croatia. [This consistency] allows us to look at cultural differences in ways of working and accept that issues identified as important to all groups might be emphasized differently in each place. So the objective was to find some commonality.

There are those in some parts of my organization who say these core principles are not strict enough and should be stricter. Others say we are stating the obvious, so why even bother?

We bother because these core principles have allowed a dialogue across countries, sites, and cultures. As an example, we require an ethical review process at each institution where animal work is done on behalf of GSK. There are many parts of the world where an ethical review process is not the norm. We are asking for that to be in place. The 3Rs are part of the discussion before animal research happens. Some principles are self-evident such as food and water and there needs to be some common sense applied to implementing them.

Now that we have put together these principles we are audited to make sure we are complying with them by internal auditors who have little knowledge about animal research. They will ask us to prove how we meet the principles. So now we are working through the next level.

Most pharmaceutical companies, like GSK, have something about animals in research on their websites. For pharmaceutical companies there are basically two options. One is to apply engineering standards, and apply the most stringent standards of all the countries no matter where we work. In the case of GSK, the UK has the most stringent standards. So if we are doing work in Croatia, Philadelphia, or China, we will use [the UK] standards. This approach allows some comfort because it is very easy to measure and understand. However, it does not necessarily affect animal welfare proportionately as some would think.

The other option is to use performance standards, such as the standards upon which our core principles are based, and to use international standards such as AAALAC. For GSK, working in multiple countries, it was necessary to have international standards. Why do we use [an international standard]? Here are some of the reasons. It is a global approach and in spite of the numerous regulations, this system works. Obviously, it’s voluntary and confidential. It involves looking at practice, not just engineering standards in the program.

These standards require the use of professional judgment, which may make some people uneasy because of the uncertainty of the education, knowledge, or credibility of the person making the judgment. Professional judgment is the issue that usually causes the most difficulty in that one professional differs from another professional. It is said by nonveterinarians that if there are five veterinarians in a room, there will be 10 opinions. It is very difficult sometimes to get consensus and know what the right professional judgment is.

But from our standpoint, the animals benefit if we avoid template thinking. Temple Grandin, whom many of you may know, has written a number of books on animals. She is an autistic woman who has been very successful in the animal behavior/animal psychology field. She often discusses her visits to slaughter plants, where she conducts plant evaluations. She knows that when government inspectors come in they have a 100-step checklist. Temple goes in with five or 10 different things she examines from the animals’ perspective. From those few items, she can rapidly tell the state of the slaughterhouse. I believe this is one of the best examples of why professional judgment is important.

So there is a lot of oversight: Our wonderful core principles have now become a way to audit us, in addition to the USDA and GLPs. But that is acceptable in that there is constant challenge and a dialogue to help improve and refine the system.

It is also important to recognize that we and other pharmaceutical companies conduct work outside our institutions. CROs are fairly straightforward, but academic alliances and collaborations with biotech and other companies are not. So we have instituted a process by which we look at those aspects.

Many wonder if that’s the best use of resources that might be better spent in hiring more veterinarians or vet techs. However, as stated earlier, it is a balance. We can perform research because society as a whole has stated that research is important. So there is something the companies owe the public in return.

While the standards to which the company holds us may be surprising, it is also understandable that they want to build trust with stakeholders. They feel they must answer certain questions and I and my team have to provide that information.

What are my conclusions?

• Always to remember that we are all in this together. Personally, I feel that the weakest link in all our conversations is the [conversation] we don’t have. It is much more important to talk and disagree than not speaking and believing that “never the twain shall meet.”

• Standards, particularly engineering standards, should be based on science and show a clear benefit.

• Principles keep both the science and the animal in mind and should allow diversity and professional judgment.

Overcoming Challenges—Contract Research Organizations (CROs): Setting Up a CRO in a Foreign Country

Bryan Ogden

Thank you for the opportunity to travel back to the US. I have been in Singapore for four years having left an academic environment at the Oregon National Primate Research Center to help with a startup contract research organization (CRO).¹ I am going to use some of my experiences to illustrate some of the challenges, operationally, that we face as a CRO in working across different standards.

Foreign CROs face the challenge of establishing credibility. Western companies have scrutinized the different CROs that are being established in Asia. There have definitely been some bad outcomes, some disappointing facilities and programs. Sometimes that record is used to make presumptive judgments about new CROs.

I hope that most people know that Singapore is not in China, although if one were to ask where Singapore is there would be many different answers. Singapore is located at the tip of the Malaysian Peninsula. The island where we have our breeding colony in Indonesia is a one-hour ferry ride across the South China Sea. This presentation will focus on the Singapore story.

CROs face the challenge of being competitive. The pharmaceutical companies demand a certain standard and expect CROs to do things similarly to the way they are done in the pharmaceutical industry, which can be a challenge from one country to the next. If the CRO does not meet those standards, it will not get the business, and yet it has to be profitable. The general perception is that a young company in particular needs to be faster, better, cheaper.

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¹CROs provide support to the pharmaceutical and biotechnology industries in the form of outsourced research services (for both drugs and medical devices) such as clinical trial development, management, and postapproval services. This presentation focuses on the challenges of outsourcing to CROs in other countries.

The question becomes, Should the CRO vie to be better than everybody else or just acceptable, to meet the minimum standards? And what would those standards be? Should the CRO appeal to a niche market or do a broad service offering?

Most CROs realize that they will be expected to provide humane animal care and use. Certainly our CRO does not want to attract business from countries with tighter regulations with the intent of doing something more invasive or less humane in our country or in our facility. Certainly the pharmaceutical companies and biotech companies would not want to be perceived as outsourcing to Asia, for instance, so that they can bypass humane care and use standards.

There are also opportunities for preferred provider and partnership arrangements, collaborations. For instance, we have agreements with certain pharmaceutical companies to do some of their discovery work and perhaps some of the toxicology work. These kinds of arrangements occur in different countries in Asia and most likely in Europe and the US. Such studies are important to CROs for their survival and give them a core base of income. These partnerships sometimes also provide an opportunity to work together on novel model development.

However, there are challenges with different standards across different countries and cultures. The problem may not always be due to differences in, or lack of, animal welfare regulation. There may be different government agencies that affect permits related to a CRO’s activities without specific laboratory animal welfare regulations. I will give an example.

The other issue is how the regulations are enforced. Do the regulators really understand the best standard of animal care, or at least best practice?

Within a CRO, especially in a country where there is a diverse population, there are different cultural attitudes. For instance, some males will not take direction from a female superior or are less likely to—a technician who was raised in a household with a Filipina maid might not readily accept correction from a Philippine veterinarian. A CRO must deal with these and other interpersonal and cultural challenges.

There is also the issue of the status of the animals in different cultures. What is acceptable use of an animal? In some cultures animals that are considered pets in western culture are actually food.

There are ethical considerations and compliance issues as well. Does the country have a culture of compliance, or even a culture of integrity or ethical principles of integrity? In some countries Westerners are walking wallets—it is common for people in those countries to charge Westerners several times what they would charge someone else for similar services or products. It is considered a real coup for them to cheat people.

A particular concern with CROs is cultural attitude toward saving face versus disclosure. Mistakes on studies are inevitable. If the culture is focused on saving face, there may be a tendency to hide the mistakes, which could affect the data and the interpretation of the data. This could ultimately cause far-reaching

negative effects during clinical trials. It is therefore essential to understand the ethical principles about disclosure as opposed to saving face.

Work ethic across different cultures and countries is another challenge. In some countries the work pattern is for one person to do the work and several supervisors to stand around and watch. This is based on a concept of how many hours of actual work in an 8-hour workday the employer is entitled to.

Another issue concerns the socioeconomic and human living conditions of the workers when compared to animal living standards. It is difficult to persuade a worker to implement sanitation practices for primates when the worker lives in a hut with a dirt floor, blue tarp walls, and a rusty corrugated metal roof.

Pride in workmanship affects the quality of the facilities, and ultimately the quality of the care of the animals.

My company can be used as a relevant case study in operational challenges working across different standards. It started as a spin-off of Monash University in Australia, then moved to Singapore as a preclinical CRO in 2003. Between 2004 and 2005, facilities were designed, financing was obtained, facilities were constructed and then licensed to do research; in 2006, the programs were accredited by AAALAC and began to adhere to FDA GLP compliance. The facility was accredited by the OECD for GLP accreditation in 2008.

In addition to the Singapore CRO facility, the company has a breeding facility for primates in Indonesia. The Indonesian standards for animal care, or lack thereof, will be discussed in a moment.

In 2000, the government of Singapore began an initiative to develop biomedical sciences as a major hub of the economy, with the ultimate goal of becoming the biomedical hub of Asia. They appointed an advisory committee and built some “Field-of-Dreams-style” facilities. That committee was called the National Advisory Committee for Laboratory Animal Research (NACLAR). NACLAR looked at standards across different countries, including the UK, Australia, New Zealand, Canada, and the US, and decided to adopt standards similar to those in the US that would enforce a self-regulation type of oversight of animal care and use programs. They published their guide in October 2004. The Parliament passed an amendment to the Animal and Birds Act covering animals in research, effective November 2004. The Agri-Food and Veterinary Authority (AVA) was assigned a role similar to that of the USDA, overseeing the licensing and inspection of animal research facilities.

Of the challenges mentioned earlier, some result from working across different countries, but others are specific to Singapore. To meet those challenges in part, Maccine hired me, a US-trained, ACLAM board-certified veterinarian, since my career was based on the US standards Singapore was adopting. This helped provide credibility for the company to potential clients. The company also received some backing and support from Quintiles, a UK-based CRO, including some key staff. They recruited people from other countries, too, because there was not a base of study directors or technical staff with experience in laboratory animal care in Singapore.

Maccine has held the Indonesian facility to the NACLAR and AVA standards. In fact, the IACUC in Singapore oversees the animal care and use program at the Indonesian facility, including twice-yearly inspections of the facility. AAALAC accreditation was very critical. We even invited the Singapore AVA to be at the AAALAC site visit in Indonesia, even though AVA had no regulatory authority there.

Training has been a key issue. Through the Singapore Association of Laboratory Animal Science, which we established in 2004, we have offered training for IACUC members, similar to what is done in the US with IACUC 101.

The NACLAR guidelines are different in some minor ways from US regulations. While they are very similar overall, NACLAR does not reflect the difference between the USDA Animal Welfare Act and Public Health Service policy with regard to the definition of an animal: NACLAR guidelines cover all vertebrate animals and the AVA uses the NACLAR guidelines to measure compliance. Another difference is that the IACUC may not do a designated review. Also, to have a quorum of the IACUC, one of the people in attendance must be the nonaffiliated or the nonscientific member.

Animal facilities are inspected by the IACUC and AVA only once a year, although the IACUC program review occurs twice a year. To facilitate the process, we have started doing our IACUC inspections at the same time as the program review, twice yearly. This is similar to what is done in the US and what our clients expect. AVA inspection is scheduled, as opposed to being unannounced as it is in the US. In fact, the AVA has begun to require something similar to an AAALAC program description prior to its inspection. It seems that the AVA is striving to hold facilities to a standard even beyond the regulations and guidelines. It is both good and challenging to have inspectors come to the facility who have both an intimate knowledge of the program and the regulatory power to enforce the regulations.

NACLAR and the regulations under the Animal and Birds Act both require training. IACUC members must receive formal training. Also, anyone who does research must attend a course on the responsible care and use of laboratory animals.

The contrast between Singapore and Indonesia demonstrates an interesting continuum in the amount of regulation. Singapore is one of the most regulated countries with the highest standards in Asia, and Indonesia (and perhaps Malaysia) are at the other end of the continuum. Malaysia is now working on setting up some national animal welfare standards for laboratory animals.

As mentioned earlier, while Maccine has a facility in Indonesia where there are no laboratory animal welfare standards and therefore no government inspections, the company applies the same standards as in Singapore. Even though both countries are CITES members, one can get a CITES permit in three days in Singapore, while it can take weeks or months to get one in Indonesia. The timeline for CITES approval in Indonesia can depend on who you know and in some cases who you pay. In addition, both countries require import and ex-

port permits. Again, the turnaround is very fast in Singapore and very long in Indonesia.

Singapore is very diligent and efficient in both permitting and bureaucratic integrity. Indonesia still has not been able to eliminate influence peddling and under-the-table payments in order to get permits. Without payments, the waiting period can be extremely long.

There was a challenge in Singapore with regard to GLP certification. When the company was started, Singapore was not an OECD member so there was no GLP monitoring authority. In 2006, the Singapore government assigned SPRING as the GLP monitoring authority and it began auditing for GLP compliance in 2008. Singapore became a provisional member of OECD in 2007.

I would like to address a few other CRO challenges:

• financing and cash flow, especially with a startup company;

• design and construction;

• SOP establishment—starting from scratch, then training and achieving compliance; and

• good quality control and validation and quality assessment.

Those are all challenges that are met to one degree or another in different countries, in different CRO facilities.

Credibility is an issue especially if the country is viewed similarly to its neighbors. For example, some people may be under the misconception that Singapore is in China and, if they have heard horror stories about melamine in baby formula or colleagues have revealed their bad experiences in China, they may think that Singapore has the same negative issues. From my own experience, when we have been audited by pharmaceutical companies and biotech companies, the auditors are greatly relieved when they see our AAALAC accreditation.

An additional challenge is being competitive. Singapore does not have much in the way of rodent breeding and there is no commercial rodent vendor there. Within the next couple of years, the country plans to have its own national breeding center. Now, however, we have to import rodents, which is expensive, making it hard to be cost-competitive, especially in the toxicology area.

Communication is another challenge because there is a 12- to 13-hour time difference with the East Coast of the US, so we must have late-night or early-morning conference calls.

Language is not usually a significant problem because the official language of Singapore is business English, although sometimes the accents are difficult to understand. In other countries, however, there are significant language and cultural barriers.

Sourcing supplies and equipment can become a challenge as well as the function of the regulatory agencies with regard to shipping, bioanalysis, pathology. AAALAC accreditation was an important step to becoming a credible CRO

in order to meet the expectations of clients. For CROs doing toxicology, GLP certification is also necessary.

There have been some frustrations in not being able to work with architects, engineers, and contractors that have experience in building animal research facilities. Cost engineering is a problem everywhere, and unfortunately one of the first areas to be cut in building design is storage space. It is also a challenge to get quality materials and have them installed such that the epoxy floor does not come up, paint does not peel off the wall, or the walls don’t crack soon after you move in. In Asia, contractors tend to use unskilled labor. In China the laborers are uneducated ethnic minorities, and in Singapore it is the Bangladeshis, Pakistanis, or people from other undeveloped countries. Even with the best epoxy products, inexperienced workers can make a real mess during installation, which can of course also happen in the US.

Here are some pet peeves of mine working in Asia. Workers cannot get a concrete slope floor to drain evenly—one gets pooling of water and water pouring from the room out into the corridor during hose-down washing. Another pet peeve has to do with improper surface preparation of floors for epoxy. There are often not good moisture membranes. There are problems with improper mixing so that a month later feet still stick to the epoxy floor and wheels leave indents. Of course, installation and maturing are issues here. Another pet peeve is the apparent inability to match paint in Asia, which should be possible with a simple computer program. But this does not seem to happen in Singapore. If you are lucky, when a crack is patched, the workers will put a geometric design over it, like a square or a long rectangle. If you are not lucky, it will just kind of patch the crack in a different color in an irregular pattern, maybe using a different texture, maybe a flat paint as opposed to a gloss.

There does not seem to be enough pride of workmanship in many parts of Asia. Simple jobs like applying grout or caulking that should produce a smooth line result in thumbprints, blobs, or smears and the workers appear to be satisfied. It is a real challenge to find people who are qualified. There are the “sea turtles” coming back to China and other countries after working or studying in the US. Their competence in the CRO and in other environments using animals will depend on what their US experience was. In my experience in academia, foreign workers did not always get it, even after 10 years for some of them. If people like this return to their native country and are perceived as understanding animal welfare standards and ultimately become leaders in their companies, you can expect that things are not going to be harmonized.

It is common to hire workers from outside the area, especially in Singapore, where there are only 4 million people and no farms or anyone with any agricultural background. We often hire from other countries and have a large number of employees, including some veterinarians, from India and the Philippines.

Often it is most expedient, at least initially, to hire Westerners to help train employees and to help set the standard and maintain compliance. It is a problem to retain them, however, and it is necessary to make a plan for succession, so

that the people who are left behind are trained. If a CRO wants to be competitive, it is not profitable to keep hiring people at high salaries from the West. Trained workers tend to move from company to company in search of higher salaries and this may ultimately affect cost competitiveness.

Sourcing challenges, vendors—I won’t get into that. I think I’m just about out of time.

With regard to regulation, hopefully countries will follow Singapore’s example and review the standards in other countries and develop regulations that will be similar or at least compatible to those in other countries. They will certainly learn from the experiences of other countries as well as how to enforce the regulations. However, there is a concern that without a proper understanding of the science and intent behind the different regulations, there may be a tendency for a growing bureaucracy to make things more difficult. The regulators have a steep learning curve and regulatory creep is a real threat. We have seen an evolution in Singapore in the last four years, when we first started out with simple inspections, and now we are required to fill out a lot of paperwork before an inspection. Our annual reports, similar to what we submit in the US to the USDA, have become much more detailed.

In Singapore, an example of bureaucratic creep and lack of understanding of industry needs for biomedical research is the buprenorphine story. Buprenorphine is an analgesic that is commonly used in laboratory animals, but has been overprescribed by physicians in Singapore, resulting in human abuse. The Ministry of Health abruptly removed it from the market and it was suddenly unavailable in Singapore. Both lab animal and private-practice veterinarians appealed to the Ministry of Health and the AVA, and the AVA stepped in and helped make buprenorphine legal for certain veterinarians and lab animal facilities. The challenge has been to get a vendor who is willing to supply it only for veterinary use. A year later, we still do not have access to buprenorphine. However, every quarter, we are still required to submit a report to the AVA saying that we have not received any and have not used any of that which we have not received. I usually forget to file the report and receive a notice from AVA about a month after the report is due. The last notice says, “If you don’t submit your report, we’re not going to remind you next time. You will not be able to be listed as someone to get buprenorphine.” To me, this is an example of bureaucratic creep and the negative effects of not comprehending the impact of regulatory decisions.

In closing, I would like to say that I support what Margaret Landi said about the concerns surrounding engineering standards versus performance standards. I have seen too much misuse of engineering standards, to get letter-of-the-law but not spirit-of-the-law compliance—in some cases, straining at gnats and swallowing camels, if you will excuse the biblical reference. For instance, you come into a room of rodents in Asia with some very nice, expensive ventilated caging, but the paint is peeling off the walls and there is water dripping from the ventilation ducts and the room is filthy. But there is 100 percent fresh air to the rodents. It’s HEPA-filtered.

Credibility will continue to be a problem. It is important to continue to look beneath the surface to determine if what is seen is “show” rather than actual improvement in the quality of animal care and use.

I want to thank my company for allowing me to take part in this; Gary Morrow for his help, based on experience he has had in other CRO inspections; and articles by Stacy Pritt and Jayne Mackta.

I also thank AAALAC International, which has been wonderful in sending speakers to our IACUC training courses, materials for conferences, and people to speak at our conferences. AAALAC has sent somebody to support us every time we have asked and I know that they are willing to do that in other countries. AALAS has also provided valuable training materials and certification exams. They are a tremendous resource.

I thank the planning committee for inviting me to this ILAR conference. I thank my wife, especially, for leaving our five grandchildren—the number of which has now grown to seven—here in the US and spending four years with me in Singapore.

Global Issues: Operational Challenges to Working across Different Standards in Academia¹

Steven M. Niemi

Introduction

While globalization of many human endeavors has become a truism and digital communication channels force-feed us nonstop connectivity, it is important to remember that personal interactions remain invaluable. This is especially true at gatherings like this one where we address multiple scientific, political, legal, economic, cultural, and emotional perspectives on a subject of great interest to many around the world. The Institute for Laboratory Animal Research and the National Academies are congratulated for hosting this meeting to facilitate such face-to-face exchanges and I am honored to be invited to speak.

My assignment is to provide a perspective on current and anticipated operational difficulties and needs (i.e., “challenges”) in the use of animals in academic biomedical research across national boundaries, especially involving Americans. In other words, I was asked to describe how US scientists in academia grapple with the variety of national laws, regulations, standards, practices, customs (or lack thereof for any of these) when laboratory animals are involved in multiple countries, and how things may evolve over the foreseeable future. I will begin with a few definitions and a specific framework for my commentary in this field, continue by examining current drivers for transnational academic collaborations, and finish with predictions and recommendations for the next ten years.

At the most basic level, academia differs from government and industry in their respective missions. Governing is the purpose of government, and increasing the value of owners’ equity is the purpose of commercial firms. Academe, by contrast, is focused on knowledge, both its discovery through research and its transmission through teaching. These basic differences are noteworthy in the

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¹The views and opinions expressed herein are solely those of the author and not necessarily of his employer or of organizations with which he is affiliated.

context of this symposium because academic research comes with no vested authority over the citizenry, unlike government, and no expectation of near-term financial returns (if the research happens to be paid for by industry, it is industry rather than academia that is expected to translate any new knowledge into something marketable). Thus, one would think that academia should have an easier experience than the other two entities in transnational use of laboratory animals. We will explore whether that presumption is actually true.

Another important difference between academia and these other two elements of society is that the academic scientist understands that new knowledge will be scrutinized and must be verified by others before a discovery is accepted; governments and commercial firms wish just the opposite. In any event, testing the accuracy of new knowledge can be performed by anyone anywhere in the world if he or she has the requisite knowledge and resources, another detail relevant to this discussion.

For definition purposes, laboratory animal care and use will be treated as one and the same in this presentation. This avoids having to distinguish between national differences for animal husbandry and veterinary support versus differences involving actual animal experimentation. And from the public’s perception and certainly from the animal’s experience, animal care and animal use represent a continuum.

Only vertebrate laboratory animals will be considered in this presentation. This is despite my suspicion that the majority of animals used in academic biomedical research today are actually invertebrates, including but not limited to insects such as Drosophila melanogaster and nematodes such as Caenorhabditis elegans.

An Example: China

I will address only one geographical element in an otherwise lengthy equation, i.e., the growing presence of China in academic biomedical research, with apologies to other countries. This is partly because of China’s huge population, rapid rate of modernization, and lack of established regulatory oversight of laboratory animals, and partly because of its legacy of major “firsts” in mathematics, science, and engineering² and preeminence as the world’s largest economy for many centuries. Between around 1000 CE and the Renaissance, it is estimated that China represented more than 75% of global population, production, and trade.³ And for the subsequent 400+ years, China still generated 30% of global production (until the rise of the Industrial Revolution in Europe in the

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²Simon Winchester, The Man Who Loved China (New York: HarperCollins, 2008), a biography of Joseph Needham, the Cambridge University scholar who rediscovered and published extensively on centuries of Chinese dominance in these fields until its demise at the hands of neighboring countries and colonial powers in the 1800s.

³Klaus W. Wellershoff, as quoted by Lee Kuan Yew, “Asia’s Growing Role in Financial Markets,” Forbes Magazine, February 25, 2008, p. 21.

1830s), a percentage larger than that of the US in the decades immediately after World War II. [After Mao’s Cultural Revolution and Great Leap Forward, China’s output dropped to only 1% of the world’s GDP in 1979. China has since rebounded to 5% and its spending on R&D has grown 19% annually over the past 10 years, a rate more than six times higher than in the US, which is still the world’s leader in yearly R&D expenditures.]⁴

This impressive heritage gives many Chinese citizens great pride and an expectation that their country will eventually resume its global leadership in the sciences. Hence, domestic support for the pursuit of science and technology leadership will continue to be strong. Besides India, no other country comes close to matching China in this context. But because India has chosen to discourage the use of nonhuman primates in biomedical research,⁵ China is a logical focus. By contrast, addressing the differences only between wealthy countries with respect to laboratory animals and academic research is less compelling because those differences are minor in comparison with poorer nations. Furthermore, investment in animal-based biomedical research in China and other developing countries will likely accelerate.

Current Drivers and Barriers

Several years ago in his book The World Is Flat, Tom Friedman described how Tian Xu, a Yale professor and Howard Hughes Investigator, outsourced bench work and mouse studies to long-time colleagues at Fudan University in Shanghai while his laboratory analyzed the resultant data back in New Haven.⁶ Xu’s counterparts in China enjoyed new and expansive laboratories, ample federal funding, and could remain in their country rather than traveling to the US for graduate or postdoctoral training. This arrangement permitted the American side to accomplish just as much research with trusted collaborators but at a fraction of the cost of performing that research in the US. At the same time, graduate students in the partnering laboratory in China got access to cutting-edge research, with frequent exchanges of staff in both directions. [It is important to recognize how serious the Chinese government is about recruiting expatriates that historically have stayed abroad. A related trend is the dramatic increase in Chinese students receiving higher education—from 1.4% of the college-age

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⁴Philip Auerswald, “China’s quick fall, slow return to glory,” Boston Globe, August 11, 2008.

⁵A.J. Rao. Use of nonhuman primates in biomedical research in India: Current status and future prospects. In International Perspectives: The Future of Nonhuman Primate Resources, Proceedings of the Workshop Held April 17-19, 2002, National Research Council. Washington: National Academies Press.

⁶Thomas L. Friedman, The World Is Flat: A Brief History of the Twenty-first Century (New York: Farrar, Straus and Giroux, 2005), pp. 247-248.

population in 1978 to 20% in 2005, resulting in almost half a million new undergraduates, 48,000 master’s degree graduates, and 8,000 new PhDs a year.⁷]

Other scientists in advanced countries have recognized the same cost advantages, even without close personal relationships, and these types of extramural collaborations are becoming more popular. Consider that for the first nine months of 2004, 53% of the research papers published in Science and Nature from Chinese laboratories included American scientists as coauthors.⁸ When laboratory animals are involved, the cost differential may be even more striking when one appreciates the much greater investment made in each animal in biomedical research today. For example, genetically engineered mice have proven to be a critical tool in dissecting the influences of various genes in diseases and other biological phenomena. But preparing for the actual experiment of interest involving a specific combination of multiple genotypes usually requires several generations of cross breeding, coupled with genetic analysis of every animal from each generation to make sure that the breeding scheme actually yields the genetic components of interest. The result is not only a mouse with a novel and precise mixture of genes but a mouse that represents an investment of perhaps tens of thousands of dollars before the experiment is ever conducted. Cost realities like these in a tight funding environment are leading many to consider less expensive strategies abroad.

Other drivers for transnational collaborations between developed and developing countries involve access to patient populations or environmental circumstances that are not as prevalent in wealthier nations. China is not unique in this regard, but some of its ethnic minorities with their relatively narrow genetic bases, isolated living conditions, and limited diets may offer science a better means of understanding nature versus nurture in specific diseases that also afflict patients elsewhere. For example, Xinjiang Province in the northwest corner of China is home to thirteen nationalities. Here, Kazaks eat a very salty diet and have a high incidence of hypertension and esophageal cancer with short lifespans when compared to Uighurs, who eat primarily grains and fruit and have long lives with a very low incidence of cardiovascular disease.^9,10 On the other hand, there are diseases relatively widespread in China but rare in most other parts of the world. One example is hydatid disease, a parasitic infection affecting 600,000 Chinese, with an additional 60 million estimated to be at risk. Treat-

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⁷Howard W. French, “China Luring Scholars to Make Universities Great,” New York Times, October 28, 2005.

⁸Ya-Ping Zhang and Shigang He, 2004, editorial, Science 306: 1861.

⁹He Bing-Xian and Zhang Jian-Yi, “Dietary habits and longevity along the Silk Road,” in Proceedings of the Symposium on New Horizons in Preventing Cardiovascular Diseases, Y. Yamori and T. Strasser, eds. New York: Excerpta Medica, 1988, pp. 89-93.

¹⁰D. Rahmutula, et al. Angiotensin-converting enzyme gene and longevity in the Xin Jiang Uighur autonomous region of China: An association study. J Gerontol A Biol Sci Med Sci 57:M57-M60, 2002.

ment is only 30% effective, leaving many to face premature death.¹¹ Many scientists are studying this parasite-host interaction because it may provide insight into parasite immunology in general.

Regardless of the increase in transnational research collaborations between developed and developing countries, there are significant risks involved. These include fraud, plagiarism, loss of intellectual property, and perhaps even criminal acts involving theft or smuggling of protected natural resources. Most of these adverse consequences are very rare in contemporary science among wealthy countries, but lack of similar protections and an only recent respect for the ownership of ideas in less developed countries raise legitimate concerns.

Other postulated incentives to increase academic collaboration between scientists in developed and developing countries are not as valid, at least here in the US. These include the avoidance of regulations and activist targeting pertaining to the use of animals in biomedical research. American scientists have become accustomed to established standards of laboratory animal oversight, and their respective institutions have administrative and physical infrastructures for compliance with regulations that have been in effect for many years. In fact, the biggest academic concerns to transnational collaborative research involving lab animals involve just the opposite situation, i.e., the lack of those same quality standards and safeguards that protect the health and welfare of today’s expensive animal models in developed countries.

When coupled with an unprecedented level of scrutiny available via the Internet, the negative consequences of mere allegations of lab animal mistreatment involving a scientist in a developed country are greater than any theoretical advantage to be gained by conducting animal research in a less rigorous environment. Therefore, the biggest concerns to be resolved if such collaborations are to grow and succeed involve acceptable (i.e., Western) ethical values for and adequate oversight of animal research in developing countries. Additional concerns pertain to the adequacy of the knowledge base of local veterinarians in poorer nations with respect to lab animal biology and medicine, as well as the vested authority of those veterinarians to intercede on behalf of lab animals when (Western) limits on animal pain and distress have been exceeded. Some may claim that this smacks of cultural imperialism and imposition of one ethical standard at the expense of another that has just as much legitimacy. However, even in China, as a middle class becomes more established, pets are becoming more precious to their owners and this trend could expand to greater sensitivity for other animals.¹² As a result, cultural attitudes toward animals may, indeed, converge as living standards rise and more persons gain access to the web and other global connections.

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¹¹“Parasitic time bomb,” Scientific American, July 2005, p. 22.

¹²Nicholas Zamiska, “Chinese Unleash a New Fondness for Their Dogs,” Wall Street Journal, August 7, 2006.

Current and Future Safeguards

At least two multinational approaches to address these major concerns are currently in effect. Both are laudable, but each has limitations. The first involves the NIH’s Office of Laboratory Animal Welfare (OLAW; http://grants.nih.gov/grants/olaw/olaw.htm). OLAW is charged with obtaining official assurances that institutions accepting NIH research funds comply with contemporary standards for lab animal care and use. US institutions must submit a lengthy document every four years that details every aspect of their animal husbandry, veterinary care, occupational safety, and internal (IACUC) oversight programs, for review by OLAW staff before an assurance is approved. OLAW also expects to receive annual updates from those institutions as well as timely notification of any major adverse events affecting lab animal welfare. Foreign institutions that receive NIH research funds also are required to submit an animal welfare assurance, but that document is less stringent, to wit:

When the grantee is a domestic institution (i.e., domestic grant with a foreign component), PHS animal welfare requirements are applicable. Accordingly, the grantee remains responsible for animal activity conducted at a foreign site and must provide verification of IACUC approval. That approval certifies that the activity, as conducted at the foreign performance site, is acceptable to the grantee. The grantee IACUC may accept, as its own, the approval of a foreign entity’s IACUC; however, the grantee IACUC remains responsible for the review. Additionally, the foreign entity must complete the Statement of Compliance with Standards for Humane Care and Use of Laboratory Animals by Foreign Institutions, available from OLAW. This document certifies that the institution will comply with the applicable laws, regulations, and policies of the jurisdiction in which the research will be conducted, and that the institution will be guided by the International Guiding Principles for Biomedical Research Involving Animals. If the grantee is a foreign institution then IACUC approval is not required. The institution completes the Statement of Compliance referenced above. OLAW encourages foreign institutions to use the standards in the Guide, which is available in a number of foreign translations.¹³

Currently, institutions in 79 countries have an animal welfare assurance approved by OLAW.¹⁴ OLAW cannot impose US laws and regulations on foreign entities, and regulatory site visits to those entities to ensure compliance with the Guide and other standards are impractical and unaffordable. Thus, the

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¹³PHS Policy on Humane Care and Use of Laboratory Animals, “Frequently Asked Questions” (http://grants.nih.gov/grants/olaw/faqs.htm, last revised February 26, 2008).

¹⁴“Foreign Institutions with a PHS-Approved Animal Welfare Assurance” (http://grants.nih.gov/grants/olaw/assurance/500index.htm).

resulting discrepancy allowed between US and foreign institutional oversight of lab animals can be wide. Lapses that affect scientific data or animal welfare could be subject to disciplinary enforcement in the US but may be deemed tolerable in another country. That would result in only the US collaborator being subject to punishment from either OLAW or the public. In addition, such a double standard leaves a US scientist at a disadvantage for research funding when competing against a foreign applicant or another US institution relying on a foreign collaborator for its animal work, if all other aspects of their respective grant proposals are equal and if the first US institution is not able or willing to attain an acceptable assurance from OLAW.

The second approach in effect today is the voluntary accreditation program conducted by the Association for Assessment and Accreditation of Laboratory Animal Care International (AAALAC; www.aaalac.org), a laudable organization that is well represented at this symposium. Over 750 institutions, including academic research centers, in 29 countries are currently AAALAC accredited. But AAALAC accreditation is entirely voluntary and its deliberations and communications with applicants are confidential. Thus, there may be just as much uncertainty about the relative quality and reliability of AAALAC-accredited institutions as there can be about US versus foreign assurances approved by OLAW. However, the AAALAC approach is more rigorous than OLAW’s in that site visits are required every three years to maintain accreditation, regardless of past evaluations or parallel approval by OLAW or any other entity. Those site visits are conducted by an experienced team of lab animal specialists to determine whether the institution remains in compliance with the Guide and other applicable standards of care and use as well as pertinent national and other laws and regulations. One can envision AAALAC accreditation becoming an international standard of quality in the absence of national laws and regulations that are equal for all countries.

In the meantime, there remains a need to upgrade global lab animal welfare standards not only to establish a more fair and consistent playing field between scientists in developed and developing countries but also to ensure that lab animals used anywhere are provided protection consistent with evolving values. The often referenced International Guiding Principles for Biomedical Research Involving Animals was issued by the Council for International Organizations of Medical Sciences “as a result of extensive international and interdisciplinary consultations spanning the three-year period 1982-1984.”¹⁵ It continues to be used as an acceptable foundation to guide lab animal welfare around the world, but has not been upgraded or otherwise revised for the past 23 years. More recently, the predecessor of this conference¹⁶ and a similar one-day

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¹⁵www.cioms.ch/frame_1985_texts_of_guidelines.htm.

¹⁶The Development of Science-based Guidelines for Laboratory Animal Care: Proceedings of the November 2003 International Workshop, National Research Council.

symposium at the AAAS annual meeting earlier this year¹⁷ compared and contrasted various regulatory and cultural mores involving lab animal welfare among only developed nations. But neither gathering would or could go further to propose more modern goals for universal standards of lab animal welfare that would apply to research performed elsewhere.

In light of these circumstances, combined with the need for more modern practices involving lab animals as more transnational collaborations arise, the following objectives are proposed, listed in no particular order:

• Veterinarians responsible for laboratory animal health are sufficiently educated and trained in the biology, husbandry, handling and restraint, spontaneous diseases, and veterinary medicine of the specific species under their care before they are assigned any such responsibility.

• All laboratory mammals produced or used in research, testing, or education receive environmental enrichment unless enrichment is exempted for (valid) scientific reasons.

• All laboratory mammals produced or used in research, testing, or education receive effective postoperative analgesic therapy unless exempted for (valid) scientific reasons.

• Lethal endpoints are not permitted for laboratory mammals produced or used in research, testing, or education. Animals approved to decline to moribund endpoints are monitored frequently enough to ensure they are euthanized before they die.

• Veterinarians responsible for laboratory animal health have authority without interference or penalty to intervene on behalf of animals experiencing unapproved or otherwise excessive pain or distress.

These objectives will be presented to the American and European Colleges of Laboratory Animal Medicine for submission to the International Association of Colleges of Laboratory Animal Medicine, with a recommendation that they be adopted within five years.

Summary

We are witness to a rapidly changing environment for academic animal research in which comparable expertise and resources are available at lower costs in a more transparent and more knowledgeable global society. It is imperative that lab animal welfare standards be adjusted and universally adopted to ensure that good science and good animal care continue to go hand in hand everywhere lab animals are used.

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¹⁷“Optimal Laboratory Animal Care and Use: The Road to International Guidelines,” AAAS Annual Meeting, Boston, February 17, 2008.

Overcoming Challenges—Academia in Europe

Harry van Steeg

My presentation will focus on an overview of the animal studies going on in our institute.¹ Most of these studies are embedded in international collaborations, among others, with NIH support and grant money. Differences in standards in animal testing are encountered especially in these international projects. Here is a brief overview of the studies being conducted and the areas of interest.

There are two primary research fields. First, we are interested in developing alternative test models for carcinogenicity and mutagenicity. In particular, we are interested in the mechanisms of nucleotide excision repair, or in general genome maintenance, and p53, the cell cycle control gene. In addition, we are involved in large survival studies in models that have a defect in genome maintenance genes. In particular, what is the effect of this defect on survival and aging in these models? We encountered some differences in regulations in these studies.

Why are we interested in developing alternative tests for carcinogenicity testing? The gold standard is still the rodent two-year bioassay, which is very tedious and uses many animals—at least 500 rats and mice must be used to test one compound—and a very high dose is used, up to the maximum tolerated dose, which is totally irrelevant to human exposure. These assays also require long exposure times over the lifetime of the animals, which is two years or longer. Based on these dose-regimen protocols, these two-year bioassays often produce many false positive results. Thus, the results are not reliable, and these assays are very expensive.

Therefore, we are interested in developing alternative test models to decrease animal use and use lower, more relevant doses at lower cost. The idea was to make animal models that are more sensitive to carcinogens and in that way use fewer animals.

In our institute in the Netherlands, we developed a DNA repair-deficient mouse model, XPA, and we combined it with a p53-deficient model, which was

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¹The National Institute of Public Health and the Environment in Bilthoven, the Netherlands (www.rivm.nl).

developed here in the US. Based on our preliminary results, this is a very interesting model, and we were invited by the International Life Sciences Institute and the Health and Environmental Science Institute (ILSI/HESI) to become a member of a global initiative to foster alternative testing in carcinogenicity. In this program, there were four different transgenic mouse models—among others, our XPA/p53 model. Twenty-one different carcinogenic and noncarcinogenic compounds were tested. Exposure time was six to nine months instead of two years. For each compound tested, there were only 120 animals, with 30 animals (15 males and 15 females) per dose group for each of three doses.

In this global enterprise there were pharmaceutical industries, contract research organizations, governmental institutes like ours, and regulatory entities, like the US Food and Drug Administration (FDA) and the Committee for Proprietary Medicinal Products (CPMP) in Europe. In total, there were over 70 different partners in a big program.

The outcome of the study is that most of the known human carcinogens tested positive in our transgenic models. Some compounds were false positives, which are compounds that test positive but may not actually be carcinogenic to humans. They tested positive because of the dose regimen used in this rodent assay.

These alternative transgenic models may prove to be very interesting if used as an adjunct to the two-year bioassay, which is what is currently happening. Both the FDA and the CPMP in Europe allow the use of transgenic animals as an alternative to the mouse lifetime bioassay.

The current test for pharmaceuticals still uses 500 rats with a two- to three-year exposure. There is an alternative test with the mouse, which uses only 120 animals and exposure times of only six to nine months. This clearly is a big advantage in terms of the 3Rs concept.

The next part of this presentation will focus on the aging studies in the institute. The basic research question is, Do DNA repair systems, or genome maintenance genes, have an effect on aging in terms of survival and pathology associated with aging? The experimental design included different mouse models having one type of DNA repair defect. Every survival study used 50 males and 50 females. The controls were the C57BL/6 animals. In these aging cohorts, we do complete analysis of all the animals when they are still alive, of course. When they are dead, you cannot do autopsy on them and pathology….

In order to determine what happens during aging, cross-sectional studies were done. Samples were taken from many tissues from each of 15 males and 15 females at several time points. The time points were 13, 26, 52, 78, 91, 104, 117, and 130 weeks and the total number of animals was 200.

These are very expensive experiments that we perform in conjunction with many American groups. In doing them, we discovered that there are different standards in the animal testing. As noted already, in the European Union animal experiments are performed based on engineering standards according to local and EU rules and regulations. Animal welfare is a critical issue and, at least in the Netherlands, we are required to prepare an animal welfare book. In the US

the experiments are based more on performance standards, on scientific facts…; when projects are funded by NIH…the rules are more stringent.

In Europe, there are two different regulations. One, designed by the European Union, is the Council Directive 86/609, which is quite old, from 1986; it is currently being revised. This document was mainly based on economic and political considerations, and not animal welfare; however, in the revision there will be attention paid to animal welfare issues. This law will apply to the 27 member states of the European Union. The other document, the European Convention for the Protection of Vertebrate Animals Used for Experimental Purposes (ETS 123) was passed by the Council of Europe, which comprises 47 member states. The focus of this organization is on social and cultural cohesion. The premise of this document is that humans have a moral obligation to respect all animals. While not all the member countries have ratified this document, our country did. Therefore, in Holland we obey both protocols.

An overview of the animals used in the EU from 2005 shows that most are rats and mice as well as some cold-blooded animals. Very few nonhuman primates are used. Research on nonhuman primates is still allowed in the Netherlands, but it is not very common. In the Netherlands, about 50 percent of animal studies are for fundamental research or education; the other 50 percent are for testing pharmaceuticals, vaccines, or other toxicity tests. In total, there were about 600,000 animals used in 2006. In the EU the total was 12 million in 2005; worldwide, it was 100 million to 150 million animals, which is a large number.

Most countries in the EU have signed on to those two documents. But as has already been discussed, Europe is divided on these issues. Some member states have refined the rules to make them more stringent; among those are the UK, Germany, the Scandinavian countries, and the Netherlands. The Netherlands has its own animal welfare law as well, which was passed in 1977 and has been updated since. According to this Dutch law, I am obligated by my institute to submit all new projects. They must be reviewed by the ethical commission. When the project is granted, all individual experiments need to be reviewed as well. So there is very good oversight. (I must provide documents for the US projects to the NIH. I have an assurance based on time; mine is for four years, which is the duration of the project. We have not had a site visit by NIH.)

So what is in the additional law? Nobody is allowed to do animal experiments unless the institute receives a license from the minister of health. Scientists are obliged to ask permission from the IACUC, otherwise we cannot do an experiment. The members judge whether the use of animals is legal, ethical, [and] justified, the number of animals proposed is appropriate, measures are taken to alleviate pain, and so on. Animal studies are prohibited when there is an alternative with respect to the 3Rs—there are no exceptions. Studies with great apes are prohibited, [but] experiments with other nonhuman primates are still possible.

We have three levels of education on responsibilities for people who are involved in animal studies. We have the Article 9 officer, who is a scientist who took a course in animal studies, including statistics; this course only takes three

weeks. The Article 9 officer designs experiments, which are assessed on their scientific content by an institutional scientific committee. The study design is assessed on ethical merits by the Article 14 officer and is approved by the IACUC.

Then we have the Article 12 officer, who is an animal technician and has had a complete study on animal handling, animal anatomy, autopsy, and so forth. The duration of this study is three to four years. Article 12 concerns all animal handling, including breeding, maintenance, critical surveillance, autopsies, and euthanasia.

Finally, we have the Article 14 officer, who is an animal welfare officer with an academic education in a biological discipline and postdoctoral training in laboratory animal sciences. This person is not, per se, a veterinarian in our country, but is involved in all ethical and animal welfare issues.

We are not allowed to let the animals die when they suffer, particularly in the longevity studies we are doing. We have criteria defined as to when we need to kill the animals. Those criteria are when there is extensive weight loss (10-20% of the total weight within two weeks); when there are changes in behavior, such as lower mobility or hunched back; cyanosis; and tumors or ulcers. The final decision on euthanasia is made by the pathologist after consultation with the Article 14 officer. The Article 12 officer euthanizes the animal.

At the end of the animal’s life, we prepare an animal welfare book describing animal discomfort, if any. This report goes to the legislated inspection services for review, and information is used to adapt for future experiments.

What does this mean for aging studies? One of the parameters of these studies, of course, is determination of the lifetime of the animals. This is a crucial parameter. However, when animals suffer—for example, when they develop ulcers or tumors—they need to be euthanized…. The Netherlands does not permit keeping the animals alive under those conditions.

In housing, it is not permitted to keep the animals solitary. However, when the animals get older and older, every now and then there will be only one animal in a cage, but because that is not allowed, buddy animals must be added.

The question, of course, is, Do the animals live for shorter times in our hands? We do have one advantage in killing the animals earlier in that we can obtain more end-of-life pathology, and we can determine their causes of death. However, the survival is worse.

The survival curve of C57BL/6 female mice shows that 50% survival is approximately 110 weeks; the survival curve of the XPD female mice with an accelerating aging phenotype shows a shorter survival compared to the C57BL/6 mice. According to the literature, C57BL/6 females typically live between 110 and 115 weeks and in our lab, the survival was about 110 weeks. Therefore, even with the requirement for humane endpoints, the survival did not differ significantly from that of animals who were allowed to die naturally.

Based on these longevity studies I conclude that under our conditions we can still do reliable studies and that the studies are comparable to those of others. Euthanasia is not a restriction. Therefore, although restrictions concerning

animal welfare are more stringent in our country as compared to the US, I believe that the outcomes of our experiments are equally valid. Given the regulation, there is no restriction on the types of research or toxicology testing we do. We can do the studies we want, provided we go to the ethical commission. And our research is still competitive, given the fact that we get some financial support also from the US.

Given all of these points, I believe that we should take animal welfare into account. It does not compromise the experiment and it is beneficial to the animal.

Finally, I would like to mention our collaborators on the many projects, both nationally and internationally. I acknowledge the collaborators on our cancer studies in the Netherlands, at the University of Amsterdam and Leiden, and those on our aging studies at the Erasmus University in Rotterdam. In the US, we collaborate with the Cancer Center at MIT, M.D. Anderson, and the University of Cincinnati. We are intensifying our collaboration with the National Institute of Environmental Health Services, to develop alternative tests for the National Toxicology Program (NTP) for carcinogenicity testing. We collaborate with the Albert Einstein Institute in New York, Lawrence Berkeley Laboratory, and the University of Texas on aging studies. Our financial support comes both from the EU and from the US with three NIH grants.