Human Practices

Would you like to introduce yourself briefly?

My name is Christian Schulze Gronover, I am a group leader at the Fraunhofer IME and we have been working on isoprenoids in plants for more than 15 years. One of the main topics being the natural rubber in dandelions, which is a long-chain polyisoprenoid. And the other branch that is essentially being pursued is the transfer of isoprenoid production from plant systems into microorganisms and there we focus on yeast. What is the relevance of isoprenoids or terpenoids?

Terpenoids, e.g., isoprenoids, are one of the largest classes of substances in secondary metabolism, not only in plants but also in animals or algae and fungi. This group of natural substances has a variety of properties that can be used by humans as ingredients in the fields of cosmetics, animal nutrition, and pharmaceuticals. Therefore, it has a long-term perspective for us to be able to use natural substances and transfer them into products to help people in the end. How are isoprenoids currently produced?

There are traditional methods, such as traditional Chinese medicine, where these groups of substances are extracted from plants and mushrooms for example, and then purified or sometimes administered as raw extracts or in powder form. In Europe, and especially in Germany, the pharmacopoeia was established relatively quickly in the field of medicines, and it states exactly which substances may be used, what purity they must be subject to, which plant they must come from, and the plant is also described in great detail. Production relies heavily on the wild collection of plants, and these wild collections are becoming increasingly difficult. For one thing, these plants are no longer collectible everywhere in their natural occurrence because the climate is changing, and also the people who can collect the plants, who know the plants, so they don’t collect something wrong, are becoming fewer. This is also because it is a very poorly paid job. The qualities that you often get from cultivation systems that are mostly abroad are very different in quality because of the price situation of how much a product can cost, and they are sometimes contaminated with pesticides or heavy metals. Corresponding companies in Europe or Germany that focus on such naturopathic preparations or phytomedicines have difficulties in getting access to these raw materials. And that is a driving force for us to say we are trying to get these substances either directly from the plant by making use of suitable cultivation systems or produce them via fermentation route in microorganisms. This is often challenging, but finally you can get more stable production rates and it is less dependent on environmental influences. In addition, we can also modify and customize these substances via enzymatic catalysis. Why do you use yeast for fermentation?

Yeast has the first advantage that it has the status being considered as safe. We know baker’s yeast from brewing beer, through wine production, but also other biotechnological products have been produced in yeast for a long time. It has this “GRAS” status, so it is generally regarded as safe for processing of products. This is not always the case with other microorganisms. And we are using yeast, a eukaryotic system (compared to bacteria currently), because there we have much larger compartmentation within the cell to separate metabolic pathways from each other, to be able to keep product accumulation within compartments, that is more in line with the plant system, and that can be crucial for more complex metabolites. How would you assess the safety of the biological methods that we use, which are very similar to yours? You already said that the baker's yeast has GRAS status, but how does that look in detail?

I think it's good that you guys rely on two systems. It is obviously the case that in the end, in terms of yield, you have to look at which system will prevail. But from a risk perspective alone, you should always rely on two systems because not every enzyme works equally well in one or the other yeast. Then your systems have two different metabolic preferences. Yarrowia is specialized on fatty acid metabolism, while Saccharomyces is more of a sugar-based, ethanolic system. Both, of course, offer some opportunities to intervene in the metabolic flux. Whether the precursor molecules for these isoprenoids come from fat metabolism or from the fermentation of sugars can have advantages and disadvantages but you can only see what works better once you have tried it out. And what about safety for society and the environment?

I see no problem with either approach, because they are so-called S1 processes, e.g. no risk for humans, animals, the environment, according to genetic engineering or risk assessment. Since the process is contained, if you think in the direction of your laboratory work or later on production, sealed vessels and fermenters will be safely filled and emptied. You can also then ensure inactivation of the organisms used or produced there. These are usual processes that are carried out in large-scale biotechnological industry on a daily basis. Thank you so much for taking the time!

You are welcome, I find it exciting and you do a great job!

Could you briefly introduce yourself and your work?

My name is Gabi Lobinger. I have been working in forest protection since 1984 at the Forest Research Institute (FVA), which is now the State Institute for of Forestry (LWF). I studied Biology in and of itself and did my diploma thesis on insect viruses at the FVA, as I always found biological and biotechnical control methods exciting. That is why I came to the FVA, now the LWF, to actually have practical relevance and direct application possibilities. Since then, I've been involved in day-to-day monitoring, pest monitoring, development of forecasting methods, routine diagnosis, on-site diagnosis, control measures if needed, and development of recommended actions for forest owners and forest management. In terms of research, my main focus is on bark beetles and oak pests/oak plant communities. There are also some joint projects with several state institutes and universities, and we constantly have follow-up projects where we work on various research questions. Here at the LWF, the reference is more practical, e.g. with very clear questions from practical work, which we try to answer with basic research and then develop corresponding recommendations for action.

What are the current concrete recommendations for action, e.g. is there a standard?

Yes, there are catalogs with recommendations for various damage situations. You are mainly interested in the bark beetle. There is a lot of material on our website, for forest owners, as well as scientifically in-depth information for the forest administration and research reports from our side. There is already a standard: until now the classical bark beetle control is the clean forest management. Despite all the other experiments and research, we do on the side, the method of choice is still to chase the beetle. It means, we always have bark beetle damage first and do even more damage by controlling it by felling infested trees and producing clearcuttings. But it is the only way to contain mass propagation, and until now there is unfortunately no other method. The use of pesticides is out of the question, as it is not recommended. The only point at which pesticides (insecticides) can be considered is a polter treatment. This happens when the infested wood has to be left in the forest e.g., because it is not possible to drive the wood out of the forest for logistical reasons. Aside from that, there is no chemical bark beetle control in the sense.

Among other things, you have researched verbenone, which is a biotechnological approach to control, can you first tell us what that is exactly?

It would be a biotechnological control because it is part of a biological mixture of substances. Verbenone is a form of communication among the bark beetles, a chemical respectively olfactory communication. This control method would be specifically related to the spruce bark beetle since it is our most important enemy in the coniferous forest. We still do not know how the first bark beetle finds the tree it attacks. Sick or damaged trees do not give signals for the bark beetle to attack them. Unfortunately, there is still a fraction in teaching that holds to this. Meanwhile, it has been scientifically proven many times that the principle is "trial and error". The bark beetle flies to a tree, determines that it is okay by scanning the bark structure, among other things, and determines if it is spruce or another tree species that it likes. It does not specifically target damaged or chronically sick trees, but trees fallen by windthrow or other damage or, if local density of beetles allows it, rather healthy ones. This is due to it wanting healthy breeding trees for its offspring and thus optimizing its breeding success. In conclusion this first signal may be a visual one or only a coincidence that concludes which tree the first beetle settles on. When they bore in, they then produce aggregation pheromones, e.g. the signal: "here is breeding space, come all". They then use this to attract thousands of males and females. It is known how this attractant is composed, and that it is produced by conversion in the bark beetle body from bark terpenes, which it eats while boring in. Verbenone is the second step: it is an oxidative conversion product of these bark terpenes. The bark beetle itself does not produce the distracting substance, but it is formed by oxidation processes over time and then gives the signal: "here is fully occupied, fly on". Verbenone has a short range, approximately only half a meter to a full meter. Through the attractants, beetles continue to be attracted. The purpose of this is to achieve a high local density. Shortly before they reach the already infested tree, they perceive verbenone and fly to the nearest tree. That is a sophisticated process, though it too has its flaws. There are trees that are so densely and quickly infested that not enough time passes for the bark terpenes to oxidize into verbenone. If this is the case, the tree is so full of larvae that they have to starve. Therefore, Verbenone doesn't perform its function in the natural context when the attack density is extremely high. Verbenone is more something that works before mass propagation or in the run-up to mass propagation. During mass propagation, the bark beetle wants to get rid of their egg stock. We have also been able to see this in studies. This connection has been known since the 1980s, and since then research has been conducted worldwide. From the beginning of the 90s, I too have been involved. Verbenone is very easy to produce, it is simply cold-pressed verbena oil. It also works very well against bark beetles, as it was tested in trials, reduce where it reduced the attack numbers up to 90%. Nevertheless it has the disadvantage that it is extremely volatile. I spent about 10 years with Bayer Crop Science trying to make microcapsules that could be aerially applied over a wide area, for example. We tried that once on eight acres, but we never got the right capsule size or capsule wall thickness to deliver the right amount of verbenone. We always had less than satisfactory trial results. When applying it to the trunk, for example, you can see that there was no infestation where it was applied, but there was infestation next to it. This is another good example of how verbenone only works for short distances, which in turn means that the only way is area application. Anything set up locally doesn't work, so it would have to be something that could be applied from the air, for example. We really pushed our research to the limit here. I just dropped out at some point, like many others. As far as I know, there are still studies going on in China and the Canadians have a paste that they offer. The last attempt of mine was a bachelor thesis on the topic in 2018 which failed as well. We simply need a distributing method with large efficiency to prevent massive damage. We must scare away at least 80-90% of the beetles. Once the infestation is there, the aggregation pheromone works, and that works 1000 times stronger than verbenone. I found it very difficult to stop this research. Nevertheless, I hope that others will be successful with it or that someone may have a brilliant idea.

So you don't think the use of verbenone in German forests is particularly realistic at the moment?

So far, there is simply no way of applying the product so that it has a long-term effect over a large area. If you apply pure verbenone, it is gone in about half an hour. It's extremely volatile and has a very high gas pressure. On the one hand, you have to make the substance less volatile, so that it can last for at least a couple of weeks; on the other hand, you have to release enough so that there is a biologically effective rate of release under different conditions, warm, cold, with or without wind, etc. Just determining what rate of airborne verbenone is needed to produce the biologically effective response turns out to be very difficult.

If it is so volatile anyway, do you know if verbenone has any effect on other insects or animals?

I think it could affect other bark beetle species as an informant since pheromones are often quite similar. But I think it certainly wouldn't have any real adverse effects. It would simply be information, which these then also perceive. Possibly this also influences other bark beetle species in the short term with the infestation behaviour. A bigger impact I do not see, especially because of the short length of stay and short range. However, this is challenging to investigate, as it is challenging to prove these effects at any rate.

In older papers, situations were mentioned such as windthrow, snowfall, and drought where verbenone could be used. Is this outdated information, or are these emergencies where it can be used?

These are the situations that we from our group of colleagues consider reasonable. Application areas could be fresh windthrow, where you want to contain mass reproduction. By preventing the infestation with verbenone, and consequently the multiplication, you can gain time for processing. The application of verbenone is only worthwhile before or in the run-up to mass propagation. In mass multiplication, it is indeed too late. It would also make sense to use verbenone in timber yards, where it is impossible to remove the wood and treat it with pesticides. Or, for example, in protection forests where you can't take out infested trees, but you could fly over them with a helicopter. These are the desired areas of application. Unfortunately, as I said, up to now there is no indication of whether it will work. It all fails with the formulation. For example, the paste that is also used in Canada is applied to the logs to protect the edge stand, which seems to be working quite well already. It used to work for us years ago, but then the material changed. The paste is not what it used to be. The release rate has changed. In the bachelor thesis, we could no longer determine any effect, except harmful effects on the trees that were treated with the paste, because it attacked the cambium of the trees. Currently, a trial is underway with the FVA Baden-Württemberg, in which the paste is dropped from the air to ensure an area-wide application. But I don't know how effective the trials were or are. The preliminary tests had efficiencies around 40-50%, which is just not enough to fight bark beetles.

So you don't see any population dynamics by applying verbenone?

No, at least not demonstrably at this point in time.

But once the efficiency is optimized, could you see the use of verbenone as large-scale protection?

Yes, that would be the big goal. The nuts and bolts are in the formulation of this substance and the release. The method of choice would be a liquid product, in microcapsules or something similar, that you embed in uric acid. That would be biologically safe, so would have no negative impact when applied. The problem, however, was ultimately the release. Will enough be released to have the desired effect, even under different environmental conditions?

Approximately what amounts of Verbenone is used per hectare?

We used about 500 ml per hectare. When you buy verbenone in the chemical store, it is sold as a small ingredient for perfumes, shower gels, etc. so it's incredibly expensive. A hundred millilitres cost over 100€, even more by now. Fortunately, as Bayer Crop Science explained to us, it is effortlessly to produce verbenone on a large scale. So far, however, there is not much demand for it. Everything sold so far is derived from cold-pressed verbena oil. It would be a source, a plant that is effortlessly to grow. The production would be cheap on a large scale.

What is your opinion on the biotechnological production of verbenone?

You do it with yeasts. I have to say, I'm not a chemist, so I can't judge how well that works. I have no problem with it though. I could even imagine it being easier to manufacture, especially for constant production. What is very important with insect scents or messengers is that the composition of different enantiomers works, that is, the isomerism. There are 1000 possible combinations. Unfortunately, the substances change simply through oxidation, for example, if you leave the bottle open too long. The insects are so sensitive that the isomer composition is decisive for the reaction. We tried many different verbenone products from the trade and thought that concentrated 98% pure verbenone was the best and most effective. It wasn't. It was a 94% product with foreign substances in it, but it had just the proper isomer composition. That's what makes even the production so insanely difficult. You can see it even now with the pheromones that have been used for a long time, like with the oak processionary moth: unexpectedly the pheromone no longer works. For 2 years we bought baits which didn't worked. We didn't know why. This was undetectable even with gas chromatography. That means the product must have excellent stability and there can´t be unwanted mutations in the yeast culture. This is for the reason that as soon as the composition changes even slightly the result can be that the desired effect is gone. That's a real problem, generally with anything that involves scent-controlled behavioural things. The big dream of most people who do research on insects is that eventually you can intervene with their behaviour even before they cause damage. Unfortunately, until now, there is always a point where something goes wrong. The important thing is quality control in order to produce a stable product with identical properties.

We can make it enantiomer specific; would that be a progress?

Yes, that would be perfect, of course. Additionally, you doing it with microorganisms, makes doing it completely large-scale easier than with plants. For example, we had a small verbenone trial, which was supposed to be part of a master's thesis based on the aforementioned bachelor's thesis, using pure verbenone. That did not work, only about 20-30% repellent effect, this means, 20-30% less trap catches of bark beetles than without verbenone. To blame was the origin of the starting products, from the verbena. It had to be a different breeding or a different genotype. This one may have been used because it's easier to grow or less susceptible to pests. Unfortunately, this changes the substance needed quite significantly. This is why the approach of making it in yeast may be a better one because you can standardize that.

Thank you very much for your time and helpful information.

I wish you all the best of luck with the production and your competition and hope that other colleagues might then have a brilliant idea on how to implement/release it. If you have more information, please feel free to share it with me as is important to me that something is happening.

Would you introduce yourself, your position and what you are working on?

My name is Johann Ach and I am the managing director and scientific director of the Center for Bioethics at the University of Münster. Primarily I deal with the whole range of bioethical and biomedical ethical issues. In the last few years, I have worked less with questions of synthetic biology, nanotechnology or biotechnology and more with questions regarding animal ethics. That's why I have to say right away that I'm not an expert on synthetic biology, if anything, I am an "expert" on bioethics.

As a bioethicist, you not only deal with ethics a lot in general, but also what values a biologist might have to or should have. Can you tell us something about that? And in your opinion, what is the responsibility of a scientist?

That depends, of course, on the areas in which you are working and what you are working on. Basically, I think it is important that biologists have an understanding that their own actions do not take place in a "morality-free space", so to speak, but that they have a moral responsibility for their own actions. This goes for various aspects: On the one hand, of course, they must adhere to the rules of good and conscious scientific work and thus fulfill their professional ethical obligation. On the other hand they have a certain responsibility for the consequences of their own actions. That means that in a certain way they are also responsible for what happens to the results of the knowledge or the materials etc. they work with. Furthermore, I believe that they also have a social responsibility, in that as biologists (if they work in science) they have an obligation to answer society's questions about what they do. In that regard they also have a responsibility to warn, if you will, if it should turn out that what they are working on creates socially relevant problems. It's a very complex issue, which requires biologists to be willing to reflect ethically, also on their own actions. This in turn presupposes that they are familiar with ethical issues, at least in basic terms, because you cannot think about ethical issues if you do not have the tools. That is why it is important that ethical questions are also made a topic in the context of biology studies, for example.

What problems can arise when students (and therefore partly not fully trained people) get access to technical tools, for example also genetic engineering tools like the CRISPR-Cas9 system?

Well, it is always a problem when people get involved with things that could create ethically relevant consequential problems without having given it enough thought. You definitely have to take precautions there. It depends very much on what (area?) we're talking about, of course, because it also depends on what kind of ethical problem, if any, we might be talking about here. Sometimes it is simply a matter of generating data that may be sensitive data and that requires a special form of data protection. Sometimes it's a matter of working with research subjects who must first consent and give informed consent before the research project can be conducted. Sometimes it's a matter of working with non-human animals, especially in biology, and having to think carefully about which ways of dealing with animals are morally okay and which are not. Beyond that, there are possible risks that may emanate from the application of corresponding genetic technologies like the CRISPR-Cas9 system. Again, these risks are very different depending on the context of application you must think about that when you working in a specific area. You have to be prepared to include a level of reflection in your own actions.

How can one deal with the fact that the project one is working on/researching may have a negative effect on communities?

I would say the first step is to be clear about what you are doing. Furthermore, it is important that you do an ethical reflection on what you are doing or what you are planning to do. I would suggest that you orient yourself on the triad of stylistic means and consequences and look at these three dimensions of criticism on your own account. First of all, ask yourself questions. "Are the goals associated with this research project goals that are ethically acceptable?" - that will be the case in many cases, for example, when it comes to developing new drugs in medical research or developing new diagnostic or therapeutic tools. In biology, it may also be about enabling certain production processes that were previously impossible or nearly impossible. These are often goals that are at least morally neutral or whose pursuit may even be ethically required. But there may also be goals for which this does not apply. That's why it's important to agree on the goals in the first place. The second dimension would be the dimension of the means one uses. One must consider whether the means one uses to pursue a certain goal are actually morally justifiable. Think, for example, of research with human embryos or animal experiments that are sometimes necessary to achieve certain results. Another example is interfering with the genome to get certain results. Some people – I personally am not one of them - think that this is also a problem and that it is at least ethically questionable when you intervene in the genome of organisms. Finally, we have to think about the risks of the application of new technologies. Perhaps already starting from the implementation of the research, we must consider whether these risks are in proportion to the possible benefits of such a project. In addition we have to think about other indirect social consequences, we have to consider what it would mean if a certain product, a certain technology, were to come onto the market and then be introduced on a broad scale. What social upheavals, if any, might result from this, and what options might be available to counteract these social upheavals. Let me give you an example: If we think about the application of genetic engineering in agriculture, we will have to consider whether a very broad application of this technology would not ultimately lead to a kind of monoculture, whether this would not further promote the concentration processes that we are dealing with in agricultural production. There will be similar societal risks in other areas as well, and we have to think about those. The nice thing about this model is that there is a cascade here, you don't have to think about the legitimacy of the means once you have decided that the goals are ethically problematic. And you don't have to think about the consequences if either the ends or the means are problematic. That's why you can discuss it in order. But I think you also have to do that with respect to virtually any technology in any research that you do.

As a researcher, how can you deal with the fact that in some situations you make use of knowledge gained through projects that were carried out under ethically questionable conditions in other countries?

This is a problem we have been carrying around for a long time in the history of science. The prime example is, of course, research during National Socialism. That alone must be bad research, because it took place under ethically questionable or ethically catastrophic, wrong conditions during National Socialism. The question is, what does one do with such results? Should one, must one ignore them, because they came about in an ethically unsuitable way, or may one nevertheless work with them? Another example for this type of scientific research that deals with these questions is research results that have been obtained through embryo research. If one has the opinion that embryos are under the protection of the human dignity guarantee of the constitution, then one primarily must ask oneself whether all the research results in some way based on embryo research not also inherited this moral problem. It seems to me that the solution to these problems is a strict separation rule. That means that one has to be very careful to distinguish between the use of the information or the results of the research on the one hand, and the extraction of the results on the other hand. In other words, one has to be careful that one's own research does not make a causal "contribution", if you will, to the practice that is seen as immoral. This can be done in various ways. One example is, that one takes institutional precautions to prevent it. However, it seems to me that the most important way, so to speak, is that those who consider certain practices to be morally unacceptable and nevertheless want to profit from the results explicitly formulate this as a problem and do everything to avoid profiting from this questionable practice themselves.

Who decides which research project is ethically feasible and should the general public have a greater say?

That again depends on what we are talking about. We have a differentiated research landscape, and in this landscape we have implemented ethics tools time and again. If you want to do human research today, you have to go to an ethics committee with your project application and you can only carry out this research project if you have the approval of the relevant ethics committee. In the case of animal experiments, you have to obtain approval from the responsible authority, and prove that you have sufficiently demonstrated the ethical justifiability of your project. In addition, in many areas of research funding, institutions also pay attention to the ethical implications of certain research projects. If you have ever written a GG application, you will find that you come across a lot of ethically relevant details that you have to explain. So, we already have a whole series of ethical reflections that have been implemented in various places. Furthermore, we also have a public, societal discussion on ethical issues, and we have created institutions for this as well, for example the German Ethics Council. They of course cannot dictate anything, but they can at least help shape the moral landscape regarding ethical issues and make them more transparent. Unfortunately, all this does not relieve the individual researcher. Ultimately, the moral responsibility for their own actions, remains with the researchers, with the applicants. This means that there are many institutionalized forms of ethical reflection in the scientific process, but this does not change the fact that the personal responsibility for one's own actions remains with oneself.

Do you see any potential ethical problems with our project, and if so, which ones?

Of course, I can't answer that off the top of my hat. For that, I would have to deal with it much more closely and take a better look at your project with more time. I would probably also have to talk to you a lot more about it. That is one of the important insights that we have seen in recent years/decades regarding science, that it is important to bring the empirical sciences and the norm sciences into a joint discussion at an early stage. We learn from each other and by learning from each other we then arrive at somewhat reasonable positions. But if you ask me this way, this is exactly where my three dimensions of criticism would start. First, I would ask whether the goal of the project is reasonable and ethically acceptable. If you use your example of implementation to say that it is about combating the bark beetle infestation, then you are definitely preaching to the choir, at least here in Münsterland. Since the bark beetle is a plague here your project seems - at least at first glance - to be a worthwhile and ethically completely unproblematic goal. In a second step, one could ask whether the means you use to deal with this problem are also ethically unproblematic. As you are talking about the fact that you want to carry out a genetic intervention or a biotechnological intervention on an organism, you can ask yourself whether this is ethically problematic or not. That might depend primarily on the moral status of the identities you want to work with. One can have very different positions on this. In the ethical debate we usually distinguish between anthropological, pathocentric, biocentric and physiocentric positions. The anthropocentrists say that humans, and only humans, belong to the moral community. The pathocentrists, on the other hand, say that all sentient beings belong to the moral community. The biocentrists say that everything that lives belongs to the moral community and the physiocentrists go even beyond that. Each position has their own good reasons. I myself belong to the camp of the pathocentric view, so I would say that the mere fact that one works with living organisms does not mean that it is morally problematic, it would be that only if they worked with a sentient or pain-sensitive living being. Your yeasts do not belong in this category according to the knowledge we have so far. Preliminary, having this in mind, I would say the means you use seem to be unproblematic. Finally, we are talking about the consequences that can be associated with the project, and of course we know that technologies like CRISPR-Cas9 are still very much in their infancy. This means that the application of these technologies is still associated with risks, risks that relate to on-target effects on the one hand and off-target effects on the other. You have to look at that very carefully, because you have to be mindful of what you are doing in the laboratory and what the consequences could be if you release it from the laboratory into the environment. In doing so, you must assess very precisely what risks you might be dealing with and what technical possibilities you might be able to use to perhaps contain such risks. Additionally you have to think about whether the risks that will remain are in reasonable proportion to the benefits that will be produced by this project.

Will ethics be harder to maintain in research in the future, with all the technical advances and possibilities?

Why should that be? Sure, critics of technology ethics are as old as technology ethics itself. One can always wonder if ethics isn't the bicycle brake on the continental airliner, so to speak. I am not so sure that the picture of a situation where progress marches off unchecked and one has hardly any possibilities to actually intervene, is correct. That situation can indeed occur if we follow a model based on the division of labor, which says that in the laboratory the scientists do whatever they want and then when they come out with some application or idea, the ethicists come in and put the "ethical justifiability stamp" on it. It is different if we have an integrated model of scientific and ethical reflection. In other words, a model that says that empirical sciences and norm sciences must go hand in hand as early as possible in research, development and application, then I believe we are in a much better position to steer the process of scientific research and development, at least to some extent, in a certain direction. I also believe that we are not in such a bad position. So far, we have a diversity of facilities, institutions, and people who think about ethical issues and who, like us here in Münster, also try to bring this perspective into the public discussion again and again. This is important, because in the end all these ethical questions concern all of us. Dürrenmatt once wrote: That what concerns everyone must also be decided by everyone together. For this, however, it is necessary that those who make the decisions - that is, in a certain sense, all of us - know a little bit about what is on the agenda and know in part what this means from the ethical perspective. If that works, and we can certainly become even better at that than we are right now, then I'm not too worried. There are also examples of bioethical reflection, for example in questions of biomedical progress, that started very early. One such example is the topic of gene therapy. Here there the bioethical debate started earlier and more extensively than the research on the topic. Another example is the debate on genome editing, which has also been conducted very extensively and broadly. Therefore, it is not the case that the ethical debate has no outlet in science. I have to say that to a certain extent, that is my business model, that is how I earn my money, and in fact I am not one of those who are always overoptimistic. But I believe that ethical considerations also have the ability to sediment in science, that is, to slowly arrive where they belong and then become effective there.

Thank you very much for the interview and all the information and for taking the time!

I wish you a lot of fun at work and every success with your project!

Hello, Professor Jaeckel. Would you like to introduce yourself briefly?

First of all, let me express my pleasure to be here with you. My name is Liv Jaeckel. I am a professor of public law, especially of technology and environmental law at the Technical University Bergakadamie Freiberg and an associate professor at the HHL Leipzig Graduate School of Management.

What role does law play in synthetic biology and research in general?

We have to differentiate between distinct legal levels. At the highest level, our constitution regulates fundamental principles. On the one hand, the constitution guarantees scientific freedom and professional freedom, which also includes entrepreneurial freedom for those who are active in the field of synthetic biology. On the other hand, the constitution protects the natural environment and animals as well as the professional freedom and entrepreneurial freedom of those who want to work without genetic engineering or synthetic biology. When it comes to research or interventions concerning human beings, also human dignity and the fundamental rights to life and health are affected. It is for the legislator to reconcile all this legally, while the constitution sets the frame. The legislator has a political margin of appreciation in order to do justice to the various ideas ranging from enthusiasm for new technologies to rejection of them. In doing so, the legislator has to respect the fundamental rights mentioned above. That means that prohibitions and restrictions of scientific experiments must be justified and proportionate. On the other hand, however, the state has to protect fundamental rights, especially human life and health and the environment, against impairments by third parties and has to provide the necessary minimum of protection.

Among other things, CRISPR-Cas9 has become an important technology, which we also used in our work. What about its legal classification and release?

Meanwhile, the European Court of Justice has found that new methods such as CRISPR fall under the genetic engineering law. There is no special legal act for synthetic biology, but research and applications in the field of synthetic biology are basically regarded as coming under genetic engineering law. However, as synthetic biology is an evolving technology, there are discussions whether the current risk assessments in genetic engineering law can do justice to the new scientific background of synthetic biology. At least, the gene technology law was adapted to some new developments. For example, the safety classifications have been revised to keep with the new gene drive method, which is also based on CRISPR.

Are there still some gaps?

In general, it can be said that many areas of synthetic biology fall under genetic engineering law, but not all. For example, the German genetic engineering law does not apply to the purely in vitro synthesis of nucleic acid fragments, if they will not be transferred into an organism. The same is true for completely artificial systems, such as an artificially produced protocell. A gray area is the import of DIY genetic engineering kits, e.g. from the USA, that do not contain GMOs yet but can be used to produce them. Even if the use of these kits is prohibited under current law, it is discussed how the import itself can be qualified. Equally exciting and not yet conclusively clarified questions concern the applications of genetic engineering to oneself for treatment. Genetic engineering law does not cover the use in human beings. But other regulations, such as the law on drugs or medicinal products, do not apply to these kinds of self-experiments either.

What role does the use of CRISPR play in the release?

The Release Directive restricts the release of organisms whose genetic material has been modified in a way that cannot be achieved naturally by cross-breeding and/or natural recombination. The directive contains an important exception for plants whose seeds are mutated by treatment with ionizing radiation or chemical compounds. These so-called traditional breeding methods are exempted from the requirements of the genetic engineering law, as these processes are considered to be safe, based on decade-long experience. Thus, we can buy the results of these methods in every supermarket. When it now comes to the CRISPR technique, the Advocate General of the European Court of Justice proposed to apply these exemptions to newer mutagenesis techniques such as CRISPR. But, as mentioned above, this opinion was rejected by the European Court of Justice. Based on the precautionary principle, the court ruled that the exceptions for the so-called traditional breeding techniques must be interpreted restrictively and do not cover the CRISPR technique. As a result, we see a major difference in the legal treatment of so-called conventional methods of breeding on the one hand and genetic engineering methods on the other hand. The decision of the European Court of Justice was an all-or-nothing judgment. It would be for the legislator to establish a more differentiated arrangement of the various techniques. This would require a new public debate on risks and benefits on a scientific basis.

What other legal regulations would we have to follow if we wanted to produce and sell our monoterpenoids?

For one thing, genetic engineering work may only be carried out in genetic engineering facilities. Of course, this applies to laboratories with the appropriate safety levels; for the large-scale production of your monoterpenoids, a facility that meets these safety levels would be necessary. Depending on the field of application of the monoterpenoid produced, regulations of medical device law, plant protection law, chemical law, or food law may also apply. This primarily involves approval procedures.

Thank you very much for your time and expertise!

I enjoyed answering your questions and wish you every success with your project!

What is special about your job?

It's more of a vocation than a job. You don't become a forester because you want to get rich, it's the work with and in the forest. We produce a renewable raw material that is almost CO2-neutral and are now helping to shape the forest of the future, which we hope will be able to cope with future climatic conditions to some extent. And being a forester is simply a nice job, you can be active outside with the dachshund, you're out and about a lot so it's fun.

Why are (German) forests so important?

It is very important, that we preserve German forests and continue to manage them in this way so that this raw material wood does not have to come from somewhere else. If we don't produce and harvest it here on site, then it comes from primary forests, from primeval forests in Eastern Europe or from the tropics. Why not do it here when it grows back and is easy to use? Of course, German forests have the same functions as forests everywhere: recreational function, nature conservation function, are filters for rainwater, protect buildings or villages from avalanches, from erosion, flood protection etc. The entire forest has a Self-protection function.

What is the European spruce bark beetle and how is it a problem for forests?

The European spruce bark beetle is our main pest in German forests, especially on spruce. If it occurs in large quantities, it ensures that the spruce trees die off. Individual bark beetles are always present, but they don't cause any problems for the trees, they dig in and are resinated. But as soon as the trees are already damaged, they no longer have a chance to defend themselves against the bark beetles. Such a bark beetle can then consequently develop in the tree, and create its brood. At some point there are so many bark beetles in the tree that the tree will die off. Here in Germany the spruce is the main tree species and the bark beetle finds it easy to migrate from spruce to spruce, causing immense damage.

What are you doing against the bark beetle infestation in the forest?

We proceed according to the principle of clean forest management, respectively we look for and find the bark beetle. It is searched for when it is drilled in the tree and these trees are removed from the stock. Clean forest management also means that we remove all brood care material from the forest. We try to get the bark beetle, the tree and all of its brood out of the forest so that it cannot infest the other trees.

How do you detect that a tree is infested?

We have forest workers, the foresters, who walk through the entire forest and look at all the trees – infested trees draw, meaning they have a red crown, the needles turn red and they partly lose bark. Another sign of infestation is drill dust at the base of the trunk, caused by the bark beetle drilling itself into the treetrunk. This is then searched for in the whole area, which requires a lot of knowledge and feeling. The infested trees are then marked and removed really fast.

Do you think it is important to inform the general public more on this topic?

It's important that the general public knows more. On the one hand, the bark beetle himself leaves traces, on the other hand we too leave traces by cutting down infested trees to protect the rest of the forest. This can lead to confusion as to why, if the general public is not better informed. Furthermore, we also work on weekends and we use harvesters to cut down the trees, who make noise, stink and create dust. Therefore, informing the general public about why we are doing it is important. Although I believe that the rural population is sufficiently aware of it, they are not well informed on the extent of the current situation. Since this year the bark beetle population was able to increase due to the persistent drought. When it is dry, the spruce is already damaged and has less resistance against bark beetles. A lot is happening in the German forests right now, and we are really having problems. There are areas where the infestation is really endangering the forest. So, it's possible that all the old trees will be gone in two or three years.

Are pesticides used for the forest/trees?

We don't do anything to the standing tree. The only principle is that we remove them from the forest as early as possible, ideally with the bark beetles. We also use pheromone traps as a monitoring system to lure the bark beetles, in order to at least know the stage of development of the beetles. The bark beetle needs four to six weeks to develop and swarms about two to three times a year. During the swarming, we need to find where he's burrowed. This monitoring takes place via these pheromone traps in order to determine where it is at the moment, how it is developing, what stage of development it is in. The bark beetle swarms about two to three times a year and we just have to get hold of it. If we don't manage to cut down the trees, bring them to the forest road and drive out of the forest within a certain time, the trees are poisoned as the very last resort. For that we have insecticides called “Attack Forst, Fastac Forst and Karate wg Forst”. These are approved insecticides. According to the list of plant protection products there are not many approved insecticides against the bark beetle. However, as already mentioned, these are only used in absolute emergencies. Using the insecticides, only the wood lying on the forest road is poisoned. Furthermore, it is important that they are compatible with bees and do not further damage the forest.

Which factors determine the continuation of the fight against the bark beetle? (economic, ecological, etc.)

We believe that we can defeat him. We have to continue, because otherwise the bark beetle population will multiply, and the development will be very rapid. Consequently individual trees or groups of trees that are infested will very quickly become entire forests that are infested.

Does profitability play a role in the management of your forest? (Or is it purely ecological maintenance?)

We are the state forests, and our motto is "sustainable economization". As an institution under public law, we manage the forest according to sustainable aspects and try to serve all other forest functions equally. These functions include the recreational-, nature conservation- and utility functions etc. In addition we are obligated to trade economically, and are not ashamed of the fact that we earn money with the forest. Therefore we do also manage the forest for economic reasons. What is important though, of course, is that the forest is there in this form at all thanks to our management. Otherwise it would all look very different and possibly also be less rich in species, less varied.

How big is the budget used for pesticides? (If possible, per hectare)

There is no real budget as these insecticides are emergency remedies. We only use it in two cases: On the one hand, we use it to poison these stacks, the lying wood, which we cannot get out of the forest in time. That was around a thousand square meters for us last year, and that costs about €5 per square meter. As I said, we don't like doing that. There are always discussions about it among colleagues. Of course, you shouldn't do that near bodies of water, in water protection areas or in nature reserves, which is a problem as the forest consists of nearly nothing else. The second variant, where we use the same insecticides, is when controlling weevils. The weevil is a crop pest […]. In principle, however, this is not done preventively, but only if an infestation is present.

What are future goals/plans related to sustainability?
We are legally obligated to manage the forest in a sustainable and exemplary manner. That means we want to maintain sustainability both economically and ecologically. Keywords for us are i.e., the four-tree concept: we try to have four tree species in the same stock. Because we don't know how the trees will react to climate changes in the future, having four different tree species, increases the chance i of one species making it. In addition we try to establish new tree species, i.e. those that are not native to the Fichtelgebirge, and see whether they perform better or have better conditions for surviving here. For example, are trees like the black locust, sweet chestnut or red oaks very drought resistant. The spruce will of course still be involved, as this is simply a defining tree species, just in smaller proportions. Otherwise, we do not manage the forest in the classic way but we try to establish permanent forest structures. We never really want the forest floor to be bare, at any time. There should always be thick, thin, large, small trees, different tree species on the same area. Resulting in a forest that is not so massively changed by the harvest.

How big is the problem of the bark beetle really? What has been done so far and what are the side effects?

The bark beetle is a real problem. The bark beetle has always existed, but this year, being so dry, shows that the problem can get very extreme. We see it now, for example, in the Franconian Forest, which is being deforested extensively. The bark beetle is capable of deforesting entire forests. We're at a point where we're afraid of not catching up. We have now found large beetle nests in various areas which were real "shock moments". You just go into a forest and then there are a good 300 trees where you say, "they all have to go, there are bark beetles everywhere". In the last few years we had less, sometimes we had ten, then five, sometimes 50 trees which were infected. This year though it has taken on an extent that we didn't know before and that is difficult to manage. This is mainly due to the drought; we have a massive dry year. It's got to be raining now, otherwise we're really worried.

Does Verbenone's approach as a crop protection agent mean anything to you? And if so, what do you think of it?

No, I'll be honest: I googled it and didn't really understand it either. I believe this is a repellent, right?
explained to him what verbenone is in detail
Would you use verbenone?

I've been thinking about how we could use that. Unfortunately I don't think that makes sense for us, because of the huge area, we have - 1/3 of the state forest belongs to the Bavarian state forests and about half of that is spruce. The question is to what extent it is realistically possible for us to use Verbenone. Another problem is, that we want to keep them in the lying wood, not drive them away. If we drive them away from the lying wood, they just go to the next standing tree. Accordingly, I don't know how much sense it would make to drive the bark beetle away, because it doesn't fly far either, only about 800m. They can't breathe when they fly and after about 800m he dies or is alternatively back in a tree. Consequently, we would have to spray the whole forest with Verbenone and that would be a huge effort. I could imagine using it in parks though, if for example, you only have two spruce trees there. Then you could drive it off permanently.

If not, what would be alternatives or how can we adapt our product so that you would use it?

It would be great to be able to spray the entire forest without it involving extreme effort. This would also be beneficial, because it is very difficult to localize the bark beetle. There are already different approaches to detection. You can use flies, some infrared spectra - the tree emits other wavelengths when it is damaged - or dogs, but they can only work 2 hours at a time, then they are exhausted. The best so far is simply the forest workers as they have a feeling for it and can detect it well. If they walk through the forest more often, then they can see when a tree changes, like loosing its needles. He can then check if it really is an infestation and mark the tree accordingly. Another idea I just got would be, that you encircle like 10 or so trees that are infested and by doing that, cut them off from other healthy trees.

What concerns do you have about using endogenous pheromones instead of insecticides?

That it is only about expulsion and the bark beetle simply flies from one expelled tree to the next tree. Another concern is how the beetles generally react to changing conditions or, for example, if they can't find another "free" tree, whether they don't just go to a tree that has verbenone in it. On the other hand I think the idea of tackling nature's problems with natural remedies is great, since these insecticides are not only harmful to the bark beetle, but also to a certain extent harmful to the environment. So please keep me posted if you get a breakthrough!

Thank you very much for the talk!