Friends and enemies of laboratory animals

Mouse animal husbandry
Olga Andreeva, "Russian Reporter" No. 28-2010
Photos: Alexey Maishev

Since the time of ancient Greece, scientists have been accused of bloodthirstiness. And their accusers proudly bear the title of humanists. Indeed, killing animals is disgusting. But for science it is necessary. We decided to figure out why science needs mice and what real humanism is. Our correspondent visited Pushchina, the only Russian laboratory animal nursery with international accreditation.

"Animals are raised here for torture and murder," says the inscription on the fence. If you go a little further, you can find out that "The hands of vivisectors are in the blood", "Animals are not slaves of people" and in general "Stop getting paid for torture."

The management of the laboratory animal nursery wisely did not paint over the inscriptions. They have nothing to do with the science they are engaged in here.

On the road at a distance, the corpse of a crushed cat symbolically lies. There are many things in life that are not related to science.

The rats did not appreciate the feat– I am categorically against the name of our institute flashing in your article, – says my old friend, a biologist who works with laboratory animals, – I don't want trouble.

– Is it that serious?

– What did you think? In Pushchina, a checkpoint with old women watchmen was set on fire, stones were thrown at them. And on Stolbovaya Street they broke in at all…

The oldest Russian laboratory animal nursery is located at Stolbovaya station near Moscow, which has been honestly serving the needs of all domestic medicine since 1929. A few years ago, some animal rights activists under the slogans "Cruelty – no!" broke into his territory and opened the cages.

Ten thousand rats were "saved". The locals knew nothing about the noble intentions of the saviors. Seeing herds of white rats in their vegetable beds, they decided that the last times had come, and called the Ministry of Emergency Situations. The rats did not appreciate the feat. Deprived of regulated feeding and climate control, all ten thousand died for a long time and painfully in the surrounding forests.

The attack on the Pushchinsky nursery took place around the same time. The nursery staff provided me with a video report about this event. About fifty young people come out of the forest with a rapid march. There are masks on their faces. Either fascists, or anti-fascists. Here they surround the nursery, throw smoke bombs at it, try to set fire to the checkpoint door and write the above slogans on the fence in several hands. The combat operation takes no more than a minute.

Together with Professor Arkady Murashev, we go to the stylish black and purple buildings of the nursery and laboratory. Against the background of a wasteland and a wild forest, the buildings seem to be something alien. They originated here, on the outskirts of Pushchina, in the 80s on the initiative of Academician Yuri Ovchinnikov. Now Arkady Murashev is their leader. Translated into everyday Russian, this means that he is a genius of scientific management. Get an international certification, which no one in Russia has yet!..

Then there was a fence. Then the appropriate inscriptions appeared on the fence.

"Let's try to defend the big science," I say cheerfully.

– And big science does not need protection, – Murashev does not support the joke. – We're doing our own thing. We are not interested in all this.

Arkady Murashev,
Head of the Biological Testing Laboratory
branch of the Institute of Bioorganic Chemistry
named after academicians M. M. Shemyakin and Yu. A. Ovchinnikov of the Russian Academy of Sciences

– Who attacked you? Green?

– Yes, they are not green at all. They arrived on buses, shouted, walked around, left with stones, then got back on the buses and left. Everything is very organized and without pathos. I think they were some kind of extremist activists. We were just practicing methods of working with the object.

– And you are not afraid of public opinion?

– And there is no opinion. There is just ignorance. That's all.

Little furry menMurashev is engaged in what is called preclinical drug testing in avian scientific language.

The mouse is his main partner and colleague in his work. He asks her, she answers. In this dialogue, the truth is born.

– What is the difference between ordinary animals and laboratory animals?

– Physiologically very much, but outwardly practically nothing. There are white, black, like dogs. Laboratory animals are specially raised for research," Murashev patiently explains. – The result of a certain selection, genetic modification – whatever you want to call it. Let's say there are cattle. There is meat, there is dairy. It is artificially derived for a specific task. So it is with laboratory animals…

– What is being tested on animals?

– That's it. More precisely, I would say this: all medicines, food additives, cosmetics, all chemical products, which a person then uses.

From the point of view of biology, mice are little men. Their genome is at least 80% exactly the same as ours. Almost nothing human is alien to mice. This is an ideal model of any of our troubles – cancer, hypertension, Alzheimer's disease and others.

For more than a hundred years, specially bred linear mice have been nobly providing themselves for basic research, and at the same time testing all new drugs. It all started, as in conventional animal husbandry, with genetic homogeneity. What is a linear mouse? This is a mouse whose brothers and sisters, children and parents have a completely identical set of genes. In natural conditions, this is impossible. In the laboratory, one or two dozen generations of closely related crossing, or, as biologists say, inbreeding, are enough - and the line is ready.

The first clean line was obtained in 1909 by American Clarence Little. The mice had a light brown color and a breast tumor. This is the so-called DBA line. It took Little five years and twenty generations of mice to breed it. It takes about the same number of generations to breed a new breed of cows. Only it will take a hundred years. That's what mice are good for - everything is fast for them.

In the middle of the last century, geneticists joined the case. Molecular biology technologies allow certain genes to be inserted into the hereditary apparatus of an animal. Do you need a mouse that will suffer from Alzheimer's disease? You are welcome! Do you need hypertension, Down's disease, liver cancer? And it is possible.

Creating a transgenic animal is not easy. First you need to make a design with the gene that needs to be introduced into the body. Then take a fertilized egg or zygote and stuff this structure there. For 40 genetically modified zygotes, only one mouse will be born. At the same time, it is not a fact that the resulting animal will give offspring in which the inserted gene will be reproduced.

But scientists are pathologically hardworking. Whether by breeding or genetic engineering, about 20 thousand mouse lines have already been created, and by 2030, according to the forecast of the journal Nature, their number may reach 300 thousand.

Sterility, sterility and sterility againI want to continue in the spirit of reporting from the fields:

"We are located on the territory of a livestock farm that supports the livestock of laboratory mice, rats and hamsters."

However, we are categorically not allowed inside the nursery. Although we are armed with the whole set of permits: requests, faxes, signatures… With such a package of official papers, you can get to a submarine or a missile base, but not to a nursery of laboratory animals. And not at all because they are afraid of the greens here.

– Don't get me wrong, – the director of the nursery Georgy Telegin comforts us, – if you are being led into a sterile box in an open dressing gown, it means that you are simply being fooled. Humans emit four trillion microbial bodies into the environment per day. You now have an average of 200 types of microorganisms on your skin, of which only fifty have been studied. As microbiologists say, the microcosm is like radiation: if it is not visible, it does not mean that it does not exist.

We are completely broken. Trillions of microbial bodies weighed heavily on our conscience. We can't go to the mice.

– What do you want to see there? Telegin is surprised. – There's nothing there. Just racks with cages – and that's it. Complete minimalism. A nursery is a box within a box. The "clean zone" is located inside, it does not touch the outer walls at any point. Excess pressure is maintained in this zone. The air there is sterile. A certain composition of this air, the air circulation rate, temperature, humidity are strictly controlled. All materials entering this block must be absolutely sterile.

Standardization as a form of humanismScientists also feel sorry for mice.

There are at least seven international conventions governing the rules of working with laboratory animals, and at least fifteen organizations that monitor this. But in Russia, the rights of laboratory rodents are fully respected only here in Pushchina (the second world-class nursery opens this year in Novosibirsk Akademgorodok).

From the sample protocol-applications for the use of laboratory animals• "I undertake that the animals approved for research in this application protocol will be used only in those procedures and in the manner described here."

• "I conducted a search to make sure that this application protocol does not contain unnecessary duplication of previously performed experiments."
• "I assure you that the study will involve the minimum number of animals necessary to obtain statistically reliable results."
• "I have taken into account and brought this document into compliance with all applicable regulations and requirements relating to radiation, chemical and biological safety."
• "I recognize the ethical standards and administrative responsibilities associated with the implementation of this application protocol, and I guarantee that all persons associated with this study will treat animals humanely."
• "I will conduct experiments that can potentially cause pain and suffering in animals and that are not planned to be relieved by anesthesia or analgesia. I guarantee that there are no alternative procedures that would allow achieving the goals of the planned experiment."

Only here you can see a modest plastic plate – the license of the International Association for Certification and Accreditation of Laboratory Animals (AAALAC).

An international certificate means that experiments with these animals are as correct as possible, and the data obtained are absolutely reliable and reproducible. Back in the 60s of the last century, scientists came to a damning conclusion: all mice, like humans, are different. If a mouse in New Zealand reacted in some way to the drug you injected, this does not mean that an English or American mouse will react the same way. For mice, this fact was clearly unprofitable. They had to die in much larger numbers.

The scientists were also somehow embarrassed. They received data that could not be reproduced. And this aroused suspicions of their unreliability. That is why the idea of certification of laboratory animals arose.

– You see, if we want to get absolutely reproducible and reliable results, we must be firmly confident in three things: in the uniformity of the genome of animals, in their health and in completely identical conditions of their maintenance, – explains Telegin.

At this place, Russian science rested. All our laboratory mice, rats, hamsters and frogs were bred the old-fashioned way, in ordinary vivariums. To get adequate results, it was necessary to create a nursery of certified laboratory animals in Russia. This is exactly what Murashev and Telegin did.

Everything is regulated here. For example, the litter in a mouse cage should correspond to 77 parameters. Its microbiological composition, the content of heavy metals and much more are monitored. The same is true with feed, water, the materials from which the cages are made, drinkers and everything with which the mouse comes into direct contact.

All people working with animals are subject to the same certification. Each experiment, the number of animals, the set of necessary operations, the devices and devices that participate in it, and so on and so on are strictly normalized. Even the feelings of the animal are regulated: pain and all sorts of unpleasant experiences of the mouse in general.

This scientific boredom, oddly enough, facilitates the fate of mice much more than the combat marches of animal rights activists. If only simply because the number of animals dying in the name of science is reduced to a minimum. It is enough to conduct an experiment once, put its results in an international database – and you can relax.

– What is the problem of bioethics in Russia? Telegin asks rhetorically. – The fact that we have to conduct a lot of experiments. If you have a non-certified animal, there is a high probability that it is a carrier of some virus that, in a normal state, quite gets along with the body. But then you entered something to the mouse, the balance was broken, and the reaction began. And you can't say for sure if it's the result of the drug or if that sleeping virus has woken up. You have one mouse showing one reaction, the other another. You can do the work very humanely, but to get a certain average, you need a lot of animals. And this is crime.

– In theory, you should just choke on orders, – I naively assume.

Telegin grimaces and looks away.

– The nursery is designed for half a million heads, but our turnover is about 50 thousand animals a year, – he sighs. – The number of customers does not correspond to our capabilities at all. Only the most advanced ones who work with Western partners order animals from us.

If the annual turnover of the Charles River company (USA), which supplies laboratory animals to biological institutes of the world, is a billion dollars, then here we are talking about simple survival. It seems that humanism is too expensive, which our science cannot afford yet.

– The rationing process, of course, is going on, – says Telegin, – but very slowly. We have about forty institutes in our country that test medicines. But only Murashev's results meet international standards. If all standards are introduced now, then 39 institutions will simply have to be closed.

Why does a rodent die– How does the mouse work?

– I ask Arkady Murashev.

– This is not for me, – Murashev resolutely suppresses idle curiosity. – This is a very difficult question that no one on mother earth will answer you. A mouse is a kind of black box that a person is playing with. The computer is arranged in the same way – a black box for some needs. A person creates some models and then asks these models questions.

The system is simple. Chemists and pharmacists are developing a certain compound that should have this or that effect on the human body. For example, to treat the flu. Perhaps, in a test tube, the new substance copes with the virus perfectly.

But how will it behave in a real organism? Will the virus kill? Will it destroy the heart or liver along the way? That's where the mouse appears on the scene – if its liver, heart, fur and legs withstand the action of the substance, and the flu will pass, then chemists have come up with something worthwhile.

At this point, like all journalists, I am told an old joke. A puppy was brought to Pavlov's laboratory. The puppy is shaking all over with fear. Barbos approaches him: "Why are you shaking?" – "Oh, I'm so scared here!" – "Don't worry, now you and I will study what reflexes are. Do you see this button? I'm going to press it now, a fool in a white coat will come and bring us something to eat."

– Therefore, no one understands who is experimenting on whom, – Murashev sums up. – A person believes that he, and the universe, maybe, itself is setting up some kind of experiment. I threw a few genes here and looks to see if they will survive or not.

– That is, you ask the mouse a question, and it answers you.

– Yes, and I'm asking a very specific question. Here I have a certain molecule, and I say to the mouse: "Here, here's a molecule for you, and you tell me what it will cause in your body and what your body will do with it."

– What is the accuracy of the answer?

– Mice give 43% confidence. That is, after the test on mice, we can say with 43 percent accuracy that the drug will act this way and not otherwise in the human body.

– And the other 57%?

– There are two large groups of animals: rodents and non-rodents. Non–rodents are all those who are higher up the evolutionary chain – dogs, cats, primates such, primates such. After checking on rodents, they check on non-rodents. If there is a rodent and a non-rodent, we can say that we have received 70% of the information about the effect of the drug.

One test usually requires four groups of rodents of ten individuals each: five females and five males. The effect of three doses of the drug is tested on three groups, the fourth group is a control group. When it comes to checking for non-rodents, three animals are enough. In total, the number of animals that sacrificed themselves for the sake of one new drug is 43 individuals. And forty of them are mice. If we take into account their huge fertility, this is about a litter of four females.

– Here you have injected the drug. What happens to the animal next?

– First we watch him, – sighs Murashev humbly. – Then we euthanize them. A special gas is injected into the mouse cage, and the animal simply falls asleep.

– That is, after the introduction of the drug, the mouse is no longer viable?

- why? Great, it's viable! But we need to check everything. It is necessary to take blood, examine each organ for pathomorphological changes. We take 18 hematological and 20 biochemical parameters only by blood. From every animal!

Stroke model, heart attack model, ulcer modelIn the company of visiting students, Murashev leads us to get acquainted with the intra-laboratory life.

Students are worried: they have already been explained that if they do an internship here, there will be no problems with employment – even capricious Western pharmacological firms will be happy to hire.

Like biblical sheep, we fearfully travel through gloomy and high-tech stylish laboratory corridors.

– This is an operational box, – Murashev demurely demonstrates his scientific wealth. – A stroke simulation operation was performed here today. Stroke is modeled like this. A special thin fishing line is pushed up to the carotid artery of the mouse. The artery is squeezed, and the cerebral blood flow immediately drops. The operation takes place under full anesthesia. Here is a device that measures blood flow.

The next room is with a huge table and a black gas cylinder in the corner.

– This is our necropsy table. Do you know what necropsy is?

The students shrug their shoulders in fright.

– We take a mouse that has fulfilled its function, for which it lived, and inject CO2 into the cell. The mouse falls asleep and no longer wakes up. Then it is completely disassembled into jars and studied from top to bottom.

Students diligently portray understanding and fearlessness.

– This is a block for studying behavior. Here is the device on which the grasping force is studied. You gently pull the animal by the tail, and it clings to the lattice with its paws. The force with which the mouse holds on to this installation is measured. It is clear that when you introduce some kind of relaxant, the grasping force decreases dramatically.

The mouse passes the tenacity test

Another room. On the table is a complex interweaving of transparent tubes.

– And here we are modeling a heart attack. The chest is opened and the coronary vessel clings. All this is done under full anesthesia, under artificial ventilation, otherwise the animal will suffocate. At the same time, we monitor the pressure, pulse rate and electrocardiogram.

The journey ends in the seminar room. Murashev, with the carelessness of a movie star accustomed to continuous admiration, pulls out a thick volume of documents.

– And now I'll tell you what we are doing specifically. This is the June catalog of our research. For example, the drug telaron. This is an old immunomodulator. Some new properties are now attributed to him, working at the initial stages of a trophic ulcer. We simulate a trophic ulcer on the foot of a rat, then give the drug and see how it affects the course of this ulcer. Here is a Swedish project with some peptides. We study them on a heart attack model. Another drug is diferidamol. It is studied on the stroke model. Well, it makes no sense to list further. Everything is clear.

Students understand everything. And I ask Murashev the last question:

– Will laboratory mice always be needed?

– Yes, absolutely. If you want to study the reaction of the human body as a whole, you must have a model of this organism. Heart, kidneys, arms, legs, fur, nose…

– So there is no alternative?

– If you want to go straight to the human trials – for God's sake. In my opinion, something similar has already happened in German concentration camps.

Portal "Eternal youth" http://vechnayamolodost.ru22.07.2010