An unprecedented outbreak of the deadly Ebola virus epidemic in West Africa, which threatens to spread to the European continent. AIDS, which destroys tens of millions of people, and other previously unknown terrible diseases of people, animals, and plants. Where do they fall on our heads? What role do the secret laboratories of the CIA and US military departments play in this?

"Can't be! Cancer is not contagious! All these are fabrications, like “conspiracy theories” or meetings with Martians!” This is how the American authorities responded to the Venezuelan government’s accusations that the great leader of the Bolivarian Revolution, Hugo Chavez, was destroyed by infecting him with a cancer virus.

However, experts believe that such a large number of Latin American leaders (and of the left!) who fell ill with cancer at approximately the same time cannot be explained by natural causes. Among them, along with Chavez, are Argentine President Néstor Kirchner, who succeeded him in office, Cristina Kirchner, Brazilian President I. Lula da Silva, who came to power after him, Dilma Rousseff, and Paraguayan President Fernando Lugo (who was overthrown during a right-wing coup in 2012). , arranged by the CIA; and shortly thereafter diagnosed with cancer of the immune system). Cuban leader Fidel Castro barely survived a mysterious bowel cancer that struck him after the 2006 People's Summit in the Argentine city of Cordoba.

Few people know that long before the brutal concentration camp experiments in German death camps during the Second World War, Americans conducted similar experiments on the inhabitants of Latin America under the auspices of the Rockefeller Institute for Medical Research.

One of the fanatics, Cornelius Rhodes, wrote to his friend in 1931: “Everything is wonderful here in Puerto Rico, with the exception of the Puerto Ricans. They are undoubtedly the dirtiest and laziest degenerates of the thieving race inhabiting this hemisphere. For public health, some means is necessary to destroy them all. And I did everything to speed up this process - I killed eight during the experiments, and infected many with cancer. There is no health insurance or social benefits here - this is admired by doctors who are free to cure to death and torture their hapless patients.”

The "Doctor" injected cancer-causing biological substances intravenously, and at least 13 patients died as a result of these cruel experiments.

In the 1950s, Rhodes became director of chemical and biological weapons research programs at the Fort Detrick Army Center in Maryland, test sites in the Utah desert and the Panama Canal, then joined the American Energy Commission, which exposed unsuspecting Americans to radioactive radiation. radiation to determine the level of “safe radiation” and the occurrence of malignant tumors as a result of these experiments.

After Rhodes' death, the American Cancer Association established an award in his name. However, in 2004, in the wake of scandalous revelations of his savage experiments, the chairman of the association, S. Horwitz, announced that the highest award for US oncologists would no longer be associated with the name of Rhodes due to the “controversial nature of his activities.”

There were a dime a dozen such scoundrels from science in the United States, and they tested almost all the infections they invented, first in Latin America (not forgetting about experiments on their own citizens). After the war, the field narrowed due to the fact that many began to turn to the USSR for medical and scientific help. But after the collapse of the Soviet Union, truly boundless prospects opened up for these flayers.

Obama has already been forced several times to apologize to Latin American countries for experiments on people in the 40s and 50s, which led to the spread of syphilis and other sexually transmitted diseases, mass infertility, and various epidemics. However, such an apology (only after the publication of irrefutable evidence!) will not revive the millions of dead and victims of US bioterrorism, nor will it lead to the cessation of such “experiments” in the future (according to the principle “if not caught, no thief”).

Since the late 60s, the accelerated development and creation of various modifications of the cancer virus began. The work was coordinated with the National Cancer Institute, which officially developed treatments for the “disease of the century,” and unofficially participated in CIA projects to use the cancer virus for military and political purposes.

Despite the ceremonial signing in 1972 in Moscow, London and Washington of the Convention on the Prohibition of the Development, Production and Stockpiling of Bacteriological (Biological) and Toxin Weapons and on Their Destruction (BTWC), work at Fort Detrick was in full swing and by 1977 it was 60 thousand liters of carcinogenic and immunosuppressive viruses were produced.

Professors R. Purcell, M. Hillerman, S. Kragman and R. McCollum actively participated in the work, who used a “cocktail” of the hepatitis B virus in combination with an oncogenic substance for experiments not only on rhesus macaques and chimpanzees, but also over American students from the Willowbrook State School for Mentally Retarded Children.
In 1971, the American pharmaceutical company Lytton Bionetics entered into contracts with a number of African countries to study cancer patients with Birkett's lymphoma, associated with the infectious Epstein-Barr oncovirus, as well as leukemia and sarcoma. It is curious that Birket's lymphoma was discovered in western Uganda for the first time after the laboratories of the US National Cancer Center, as well as other medical institutions sponsored by Rockefeller, worked there.

One of the experts, R. King, said in the 80s that specialists from the United States infected people with sarcoma in order to “isolate the genome of the virus through reclamation, hybridization, recombination of viruses, mutations and other technical techniques.”

In the Senate Church Committee hearings in 1975, Dr. Charles Senseney, who worked at the Fort Detrick laboratory, admitted that the CIA used biologically active substances that caused fleeting heart disease and cancer to destroy undesirable figures. He demonstrated samples of weapons with which the intended victims were infected. Among them was an umbrella that fired miniature darts when opened, as well as a special blowgun for shooting needles made from a frozen toxic substance. Being as thick as a human hair and several millimeters long, these needles passed through the fabric of clothing without damage and, when injected, caused a painful sensation no worse than a mosquito bite, instantly dissolving under the skin.

Among the “new products” of American bioterrorists were also demonstrated aerosols for infecting “targets” with deadly diseases after spraying from airplanes, as well as “jumping viruses” spread through insects (fleas, spiders, mosquitoes) that jump or fly from infected animals to humans. The CIA also became a “pioneer” in methods of infection: through injections, inhalations, contact with the skin of contaminated clothing, through the digestive system through eating, drinking, and even using toothpaste.

A number of experts believe that one of the first political leaders disliked by the United States to be infected with a new cancer bioweapon was the President of Angola, Agostinho Neto. He died in the Moscow Central Clinical Hospital in 1979 at the age of 57 from a hitherto unknown form of fulminant cancer. Another victim was the former President of Chile, Eduardo Frey, who openly opposed the US protege, General Pinochet. Frey died in a Santiago hospital in January 1982, contracting an unknown, fulminant illness after undergoing a routine medical examination.

So, perhaps in 50 years the CIA archives will be declassified, and the secrets of the death of Hugo Chavez and other world leaders will become known. There is such a huge amount of documentation about the use of cancer viruses by American intelligence agencies that the existence of these weapons does not raise any questions. The only question is how it was “brought in” and who was the direct perpetrator.

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“In the next 5-10 years, it will be possible to create a synthetic virus that does not exist in nature at all, and which cannot be suppressed by the human immune system; new, artificially created viruses will be inaccessible to drugs; it is useless to use conventional means of treating infectious diseases, antibiotics, vaccines and antidotes against them.” Such a sensational statement was made by the chief army virologist expert D. MacArthur, speaking in 1969 before the US Congress commissions (“Sykes Commission”), which was supposed to make recommendations on the allocation of budget funds for the army. And he asked for little - only about 10 million dollars!

Money was allocated and hundreds of researchers and experts were involved in the work. One of the creators of the AIDS virus was apparently Dr. Robert Gallo, who in 1987 even received a patent from the US Department of Health establishing his priority in the invention of a “virus that suppresses the human immune system.”

The disease escaped from laboratories and was first discovered in the spring of 1981 in California (USA). And it had nothing to do (as the Americans are trying to convince us) with Africa and “little green monkeys.”

In May 1987, an article appeared in the London Times claiming that smallpox vaccinations in Africa (initiated by "humanists" in the US Department of Health) had caused an outbreak of AIDS. And millions of people have been vaccinated! Then a similar “vaccination” was carried out in Haiti, Brazil and other countries.

Accusations of the United States of manufacturing the AIDS virus began in the mid-80s. Professor at Berlin's Humboldt University, Jakob Segal, argued that the virus is "the product of an experiment carried out in a laboratory with the aim of creating a biological weapon." In the US media, all this was presented as “Soviet propaganda.” But in the 90s, Dr. Gallo himself announced that he had tested another, “alternative” strain of AIDS, which can enter the body through epithelial cells (that is, through the skin), increasing the risk of getting the disease through spraying the active substance into the atmosphere .

Dr. S. Monteith was one of the first who, back in 1981, described the enormous epidemic potential of the new virus, the potentially catastrophic consequences of its use by the “world elite,” and also proved its artificial nature.

And this new quality has so far prevented any attempts to create a vaccine against AIDS. That is why over the years not a single effective drug has been created against this disease.

The number of people infected with AIDS is still unknown, since even in the United States the government is preventing all initiatives aimed at even an approximate count. According to various estimates, from 50 to 100 million people are infected with AIDS. Most of all in Africa - in some countries (Uganda, Kenya) more than 50% of the population suffers from this terrible disease.

It is believed that about 40 million people have died from AIDS to date - almost the same number as died during the Second World War!

* * *

According to the World Health Organization, more than 600 people infected with Ebola have already died in the west of the “dark continent”.

The current outbreak of the disease has become the largest in the history of medical observations.
In Nigeria, Liberia and other African countries, special cordons are installed at the borders, and doctors carefully monitor all those entering and leaving. Ebola fever is considered a deadly disease to which humans, primates and pigs are most susceptible. There is no vaccine for it.

The epidemic began in Guinea in March of this year. To date, the disease is spreading to new territories in Sierra Leone, Liberia and Mali. There are fears that it will spread not only throughout West Africa, but also penetrate into Europe.

It is curious that in hotbeds of the epidemic, cases of attacks by local residents on the offices of the international organization Doctors Without Borders have sharply increased. Local residents accuse doctors of bringing the virus to the region. There have been massive protest demonstrations against African governments that are doing nothing to correct the situation.

The pogroms of the offices of a “respected international organization” are presented in the Western press as examples of “irrationality and absurdity.” Moreover, “Doctors Without Borders” extol their ethical principles in every possible way, assuring that they are “always close to the victims.” But isn’t it their own victims, as the “unreasonable” Africans believe?

Why don't Western doctors stubbornly leave Guinea, Liberia, Mali and Sierra Leone? After all, these countries are engulfed in the chaos of civil wars and conflicts, in which European countries and the United States are taking an active part. France alone has spent hundreds of millions of euros on military operations in Mali.

Everything - to restore colonial power in western and northern Africa. And it is these territories that are “cleared” of the local population during epidemics of Ebola and other infectious diseases. Moreover, surprisingly, only local residents suffer, but not the “peacekeepers” from France.

And “medics without borders” do not transfer medicines and equipment to local authorities and do not leave the conflict zone. This is precisely what gives local residents good reason to suspect foreign “aesculapians” that they are the ones spreading new strains of infection among Africans.

According to many experts, new “ethnic” weapons are being tested there, which act selectively - only on Africans. But apparently there are modifications for other racial and ethnic groups. In 2006, one of the leading American virologists Eric Pianka, speaking at a ceremonial meeting at the University of Texas, said that with the help of a new strain of Ebola fever (in his words, “with fantastic lethality”) it is possible “for the benefit of the planet” to reduce humanity by 90 %. The American virologists present in the hall unanimously stood up and gave him a standing ovation...

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Since the 70s, accelerated development of “ethnic weapons” has been carried out in the United States. And as many experts believe, new strains of deadly viruses have now been invented that can only spread in a certain ethnic environment.

Thus, “SARS” affects most of all the Chinese and residents of Southeast Asia, Ebola and AIDS - Africans. Israeli scientists are trying to create a similar biological weapon directed against the Arabs.

The British Medical Association recently stated that “progressive developments in genetics could lead to ethnic cleansing on an unprecedented scale in the coming years.”

The idea of ​​establishing “biological domination over the world” is no longer maturing not only in the minds of insane cannibal virologists, but in the calculations of politicians, military strategists, and experts! Thus, this idea was recently voiced by respectable neoconservative US politicians in the report “New Frontiers for America’s Defense.”

It says that, of course, military domination over the world must first of all be ensured by ballistic and cruise missiles, radio-controlled aircraft (“drones”) and submarines, and satellite weapons. But, along with this, “over the coming years, the art of warfare in the air, on land and at sea will be completely different from the current one, and battles will be fought in new dimensions - in space, cyberspace, as well as in the intracellular and microbial level." And further it is said that “advanced forms of biological weapons, which will select certain human genotypes as targets, will be able to bring this direction from the world of terror to its rightful place among politically justifiable means”!

* * *

The US authorities have well learned the lessons of the Manhattan Project, in particular, the transfer of data on atomic weapons to the Soviet Union by the world's leading physicists. American scientists did this not for money, but based on a sober assessment of their government, which would not hesitate to bomb the USSR and all other potential competitors on the path to world domination.

Therefore, the developers of new viruses are now subject to the most stringent rules for eliminating “undesirable witnesses.” The mortality rate among them is tens of times higher than the statistical average.

Independent American experts counted more than a hundred “mysterious” deaths (in plane and car accidents, from “unknown” diseases, “accidents”) among virologists and microbiologists working under contracts for the CIA and the Department of Defense.

In 2001, immediately after the explosion of the “pancake towers,” all Americans were alarmed by the news of letters containing anthrax spores that were sent to the editorial offices of magazines, newspapers, TV companies, and political figures. 17 people became infected, five died. These letters were the main reason for the political turn that directed US aggression against Iraq. Al-Qaeda faded into obscurity, and all media reported that “the largest biological attack in US history” was organized by Saddam Hussein.

When this twist was cemented (and later used to accuse Hussein of developing biological weapons, which became one of the arguments for the invasion of Iraq), it quickly became clear that the strain of the virus could only be obtained from the CIA laboratory at Fort Detrick. There they found a “weak link” - virologist Bruce Ivins, who, being a devout Catholic, often complained that he did not like his work for religious reasons. And in July 2008, he allegedly committed suicide by swallowing potent drugs. After this, the FBI pointed to him as a “crazed terrorist” who sent out letters with infection. No autopsy was performed, there was no investigation, and the case was quickly closed.

Interestingly, he repeated the fate of one of the leading microbiologists of the 50s, Frank Olson, who also worked with anthrax and tendered his resignation from Fort Detrick, not wanting to participate in the development of lethal weapons. And a few days later, in November 1953, according to the FBI report, “in a state of nervous breakdown, he jumped from the 10th floor of the Pennsylvania Hotel.”

One of the most famous cases was the “suicide” of the largest British bioweapons expert, David Kelly. He visited Iraq dozens of times as part of various UN missions for inspections. After the invasion, he made a sensational (first!) statement that all the “documents” about the presence of S. Hussein’s chemical and bacteriological weapons, presented by the US and British authorities at the UN and which served as a pretext for the war, were “crude fakes.” He was summoned to parliament, where during the hearings he was essentially not allowed to open his mouth, attacking him with reproaches and accusations.

A few days later, on July 17, 2003, he, as always, went for a morning walk, and his body was discovered the next day a mile from home. The official report stated that he committed suicide by swallowing 30 sleeping pills and then cutting a vein in his left wrist with a knife. But the ambulance doctors (apparently not knowing about the “order”) noted that there was no blood under the corpse. Consequently, Kelly poisoned himself, cut a vein, and then, bleeding, he himself got to the place where he was found!

In the United States, one of the most notorious events was the plane crash in March 2002, in which Stephen Mostow, a leading virologist who worked at the Colorado Medical Center, died. He was called "Mr. Flu" because he mainly specialized in this disease.

Among the dead were many people from our country who, for various reasons, went to “seek happiness” in the West. The most noticeable was a “heart attack” in 2001 in microbiologist V. Pasechnik, who was in enviable health. The West used him (like many other Russians) 200% - both as a specialist and as “an exposer of the Kremlin’s terrible conspiracies against the United States and the entire free world.”

In 1989, he went to England and worked there in one of the virology centers. Along the way, he made money by telling stories about the Soviet “binary biological weapon” called “Novichok”, that all known viruses had long been mastered in secret KGB laboratories, and new ones had already appeared. They can cause "monstrous diseases" such as multiple sclerosis and arthritis in unsuspecting Americans.

These horror stories were useful because they provided an excuse to squeeze out budgetary funds for “biodefense” (in reality, for the development of new deadly strains). But then they decided that the talkative Pasechnik talked too much about the virology center in Sailsbury, where he worked for 10 years, and sent him to another world...

* * *

“Putin’s rocket”, “hand of Moscow”, “Putin, you killed my son!” - Western magazines and newspapers were full of such headlines after a Boeing passenger plane flying from Holland to Melbourne was shot down over the skies of Ukraine on July 17 this year. This hysteria began immediately after the speech of US President Obama, who said that this was a “crime of unimaginable proportions” and blamed Russia. Immediately in the hands of the press secretaries of the White House and the State Department, some blurry photographs appeared that were received from the CIA and “irrefutably indicated” that the airliner was shot down by a Russian Buk missile.

This event served as the reason for the full-scale deployment of economic sanctions against Russia, the involvement of EU countries (before the disaster, they hesitated whether to support the United States), the use of almost all prohibited means of war to suppress resistance in Novorossiya (including phosphorus bombs, ballistic missiles, cluster warheads etc.), the implementation of plans to put together an anti-Russian military bloc with the participation of Ukraine, Moldova, Poland, Georgia, and the Baltic countries.

Only a month later, materials began to appear that holes in the cockpit and fuselage proved that the plane was shot down in the air, most likely by a Ukrainian Air Force fighter. This version is confirmed by a sharp change in the Boeing's route immediately before the disaster. However, the deed has already been done, all Western media immediately forgot about the plane, and sanctions and a full-scale war against the Russian people in eastern Ukraine are not only in effect, but continue to intensify.

There are all the signs of a “trigger event” or a “falsified incident” (false flag incident) - this is how the masters of provocations from the CIA call terrorist attacks that are designed to turn public opinion in the direction necessary for the United States, to set off a chain of events that will lead to the realization of goals "empire". This has always been the case in US history - the explosion of its own battleship Maine, which became the pretext for declaring war on Spain in 1898; the planned sinking of the passenger steamer Lusitania to enter an advantageous moment into the First World War; deliberate suppression of information about the impending Japanese attack on the American base at Pearl Harbor in 1941 to enter the Second World War; provocation with the shelling of the American destroyer Maddox in the Gulf of Tonkin to declare war on Vietnam in 1964; the bombing of the Twin Towers in 2001 to begin the “War on Terror” and prepare for the invasion of Iraq and Afghanistan.

As often happens in such terrorist attacks, not one, but several goals are pursued. In this case, of great interest is the information that there were more than a hundred microbiologists on board MH17 who were flying to the international AIDS congress in Australia. And among them is J. Lange, a leading virologist at the University of Amsterdam.

“The irreparable loss of the greatest visionary and titan in the study of AIDS,” “the tragic death of the world’s leading specialist in the treatment of the disease of the century,” were written in obituaries published in scientific journals. And, indeed, Lange’s laboratory took a leading position in the study of AIDS and methods of its treatment, including the combined use of drugs, antiretroviral therapy, and developed ways to prevent transmission of the virus from mother to child. For several years (2002–2004) he headed the international organization to fight AIDS. Along with him on board were his Dutch colleagues Jacqueline van Tongeren, M. Adriana de Schutter, L. Vann Mens and other scientists. It is possible that they brought with them the results of many years of work, maybe even a long-awaited cure for this monstrous disease - after all, shortly before the conference, Lange’s employees said that his speech should create a sensation in the scientific world.

In the same Boeing (allegedly, by fateful coincidence), was flying the representative of the World Health Organization (WHO), Glenn Thomas, who was “fined” by giving an interview where he mentioned the criminal role of his organization in the spread of the Ebola epidemic in West Africa.

By destroying European AIDS researchers, as well as an honest WHO functionary, the Americans thereby taught a lesson to all those who sincerely make efforts to cure AIDS and Ebola: “There is no need to treat and prevent these diseases, they are very useful to us for the destruction of the proliferating human rabble.”

It is no coincidence that a number of articles recalled that in 1998, a Swissair plane crashed over the Atlantic, carrying one of the brilliant AIDS researchers, Jonathan Mann, and his wife M. L. Clements, also a famous virologist. Mann headed the WHO structure designed to combat AIDS, and, as his colleagues wrote, his death dealt a powerful blow to all plans for organizing the fight against this terrible disease. The causes of the disaster have not yet been clarified (none of the serious experts believe in the official version that one of the pilots’ cigarette butt fell and this caused a fire in the interior of the plane).

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The United States uses a huge arsenal of bioweapons against us: GMOs and transgenic plants and organisms (many of which, according to Western experts, cause suppression of the immune system, cancer, infertility and brain diseases), annually organize dozens of epidemics of new influenza viruses, animal diseases (“swine” and “bird flu”), plants, spread various allergic diseases, sell medicines and vaccines with “side effects” unknown to us, food additives, etc. More and more new viruses are being developed: the deadly “hantavirus”, the recombinant “Australian killer virus” based on smallpox, a new generation of “non-fatal” (only completely “incapacitating”) diseases, “bioregulators” capable of creating depression on a massive scale, changing heart rhythms, and leading to insomnia. It is possible that biological “bookmarks” are created - latent viruses that should be activated after a certain time.

American military biological laboratories are being created around Russia: in Georgia (where, according to experts, the swine fever epidemic originated in 2013), Kazakhstan, Kyrgyzstan, and the Baltic states. The US authorities allocate huge amounts of money both for the development of new viruses and for biodefense (more than $6 billion is spent annually on the Bioshield program alone).

In our country, after the collapse of the Soviet Union, for a long time almost no attention was paid to this most important area of ​​protecting the country. Institutes and centers closed, young specialists left for the West. Only enthusiasts and elderly scientists remain who work for meager salaries (18 thousand are senior researchers, 27 thousand are professors, doctors of science).

Dilapidated buildings, outdated equipment, “additional pressure” from liberal officials. It got to the point that in 2000, for “underpayment,” Chubais’ Mosenergo tried to turn off the electricity at the Ivanovsky Institute of Virology. Not only would a unique collection of microorganisms be destroyed, but some of the virus samples could escape into the atmosphere! Then it was only by miracle that we managed to fight off the “effective managers.” And the final blow was dealt by the “reform” of the Russian Academy of Sciences - in fact, its liquidation and the transfer of management into the hands of an “effective” accountant from Krasnoyarsk.

No one interfered with the real hunt of CIA agents for patriotic scientists, who were simply destroyed on the territory of our own country! In January 2002, A. Brushlinsky, corresponding member of the Russian Academy of Sciences, director of the Institute of Psychology, psychologist and biologist, author of works on recognizing terrorists, was beaten to death with baseball bats (so that they knew where the order for liquidation came from!) and strangled in the entrance of his house in Moscow. Two years after his death, his deputy, Professor V. Druzhinin, was killed.

In November 2002, Professor B. Svyatsky, a specialist in childhood infections from the Russian State Medical University named after. Pirogov. Corresponding member of the Russian Academy of Medical Sciences, a leading virologist and microbiologist, bioweapons specialist L. Strachunsky, was beaten to death with baseball bats in 2005 in his room at the Moscow Slavyanka Hotel. In 2006, geneticist and biologist, corresponding member of the Russian Academy of Sciences L. Korochkin was killed.

A huge loss for domestic microbiology was the death of the head of the Department of Microbiology of the Russian State Medical University, Professor V. Korshunov, one of the world's leading virologists, a recognized specialist in biological “anti-weapons”. The 56-year-old scientist was beaten to death by “unknown hooligans” in 2002, a few days after the publication of a newspaper article stating that the scientist was on the verge of the greatest discovery - a universal vaccine against any bioweapon! As a result of Korshunov's death, work in the most important area of ​​science was stopped. Hundreds, if not thousands of people in Russia were doomed to death due to the stoppage of research.

The tragic pages of modern history convince us that the United States is capable of any, the most barbaric and criminal actions in its manic desire for world domination. It is significant that the countries where they invade under the pretext of “protecting human rights,” “humanism,” and “democracy” become not only the scene of the most acute civil wars, but are also accompanied by epidemics of various new, previously unknown diseases. Huge masses of people in Vietnam, Yugoslavia and Iraq were exposed to mutagenic substances, which led to terrible consequences. Terrible deformities among babies, the creation of a whole generation of degenerates, irreversible changes at the genetic level that will affect all future generations - these are some of the consequences of “humanitarian actions”.

Moreover, international organizations, including the UN, currently under full US control, play the role of “cover” in the implementation of this genocide. The World Health Organization (WHO), Doctors Without Borders, and other previously authoritative bodies write their “objective reports” under the dictation of the West, and they can no longer be trusted. They acted in conjunction with the aggressors in Iraq, Afghanistan, and Libya.

On the eve of the US invasion of Iraq, they obediently concluded that Saddam Hussein possessed “huge stockpiles of biological and chemical weapons,” which served as one of the main arguments for the US to start a war. Last year they accused the Syrian government of using chemical and biological weapons against its people when about 300 people were killed in August by sarin nerve gas in a Damascus suburb. Although by that time, strong evidence had been received that sarin was used by militants from al-Qaeda, and it was obtained not from anywhere, but from American warehouses.

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The ruthless destruction of competitors and, in fact, biological tyranny of the United States destroys the sovereignty of the peripheral countries of the world, forcing them to rely on help, expertise, and medicines from abroad. Such colonial dependence undermines the security of peoples, makes them hostages of the West, “lab rats” for various medical and biological experiments directed against their health and life.

The only counterbalance to the bioterror empire can be the rejection of vicious “globalism” and the construction of a multipolar world. All countries must, step by step, refuse cooperation with the United States and NATO, the existing pro-American international organizations. It is necessary to conclude agreements at the interstate level. Thus, in Africa, states must work together to combat new introduced strains of Ebola. In Southeast Asia - against the most acute new syndrome of “SARS”. It is at the national level that we need to take care of our science, create our own national institutes and laboratories, powerful scientific centers to counter viral and genetic weapons.

Nikolay Ivanov

Stages of laboratory diagnosis of viral diseases. Setting up a virology laboratory.

1) Indication (detection) of a virus in pathological material:

Express methods:

a) Detection of virions:

(1) Electron microscopy;

(2) Light microscopy (smallpox);

b) Detection of viral Ags in serological reactions (RIF, ELISA, RSK, RDP, RNGA);

c) Detection of inclusion bodies using light and fluorescence microscopy;

d) Detection of viral nucleic acids using PCR and DNA probes;

e) Detection of hemagglutinins in RHA;

f) Detection of the infectious activity of the virus in a biosample.

2) Isolation (isolation) of the virus from the pathogenic material. Isolation is carried out regardless of the results of the first stage using a biological test in three “blind” passages. Passage is the infection of a living system in order to obtain a new population of the virus. "Blind" passage– infection without visible signs of virus reproduction. After three passages, the virus accumulates in the cells of a living system, which is accompanied by the appearance of signs of reproduction, visible at the level of the macroorganism. For example, when biological test on animals: clinical signs, death, pathological changes; on chicken embryos– death, pathological changes, hemagglutination; on cell culture– CPP, hemadsorption, plaques, RIF, etc. Such infected living systems are defined as a positive bioassay. However, it is not yet possible to accurately determine the type of infectious agent. Therefore, pathological material is selected from a positive biosample, which is conventionally considered secondary, i.e. selected from a living system with signs of a positive bioassay. A virus-containing suspension or fingerprint swabs (isolated virus) are prepared from it.

3) Identification (determination of the type) of the isolated virus in serological reactions or by PCR analysis. In rare cases, identification by other characteristics is possible, for example by intracellular inclusions (Babes-Negri bodies for rabies).

4) If necessary, evidence of the etiological role of the isolated virus. For this purpose, serological reactions are used, in which the isolated virus is used as an antigen and paired samples of blood serum in two-fold serial dilutions are used as antibodies. A positive result, proving the etiological role of the isolated virus, is an increase in antibody titer in the second blood serum sample by 4 or more times compared to the first.

5) Retrospective diagnosis. For this purpose, paired blood sera taken at the recovery stage are used, which are tested in serological reactions with a standard specific antigen in accordance with the preliminary diagnosis of a viral disease. An increase of 4 or more times in the antibody titer in the second sample compared to the first indicates an active infectious process occurring in the animal’s body during the period of blood collection. In this case, the disease is caused by the virus to which an increase in antibody titer has been established in paired sera.

Setting up a virology laboratory.

To organize a diagnostic laboratory, use an isolated compartment consisting of at least 5-6 rooms.

A bright room is allocated for the laboratory. Rooms for working with viral material should be well lit and consist of a pre-box and a box, separated by a glass partition with doors. Only tables, chairs and work accessories are placed in the boxes. The surface of the tables is covered with stainless steel, plastic or glass, and bactericidal lamps are installed above the working surface. A rubber sponge disinfectant mat soaked in a disinfectant solution is placed at the entrance to the box. The pre-boxing room contains sterile clothing and equipment appropriate for the purpose of the boxing. The laboratory is provided with cold and hot water and ventilation with a supply of sterile air.

To register incoming pathological material, it is intended reception, where several tables upholstered with galvanized sheets and containers with disinfectant solutions (3% chloramine, sodium hydroxide or 5% phenol) are placed.

In the material pre-processing room ( opening) open the corpses and select material for further research.

Box rooms are equipped depending on their purpose.

In an autoclave, dishes, culture media, equipment, and culture media are sterilized and infectious material is neutralized. It is necessary to have two autoclaves: for clean materials and for infected ones.

The washing room is designed for washing dishes, equipment and appliances.

The vivarium must have a quarantine department, rooms for healthy and experimental animals and utility rooms.

For any type of virology laboratory, a mandatory part of the laboratory is a tabletop box or, better yet, a box with laminar air supply.

46. ​​Cell cultures and their types. A system in which cells, tissues or organs removed from the body retain their viability for at least 24 hours. Surviving: in which cells only retain their inherent life activity without reproducing. Growing: retain their inherent life activity and are capable of proliferation. According to the nature of growth, they are divided into 3 groups: suspension; plasma (cultures of fixed pieces of tissue); single-layer. Single-layered ones are divided into 4 groups: primary trypsinized; subcultures; semi-leapable and interleavable. Suspension: grow in the form of suspensions, cells multiply with a special medium plus constant mixing using rollers. Cells grow all over the surface of the mattress. A large number of cells for vaccines. Plasma: pieces of tissue fixed by plasma, this is tissue culture. It is obtained by fixing a piece of tissue on a glass virological glass, then adding pit medium and culturing it; in this case, cell growth is recorded along the periphery of a piece of tissue. Used to obtain pieces of fabric. Single layer: To indicate a virus. It is obtained from tissues or organs by treating them with trypsin. Subcultures are obtained from primary ones by grafting. Next, semi-transplanted by multiple transplantations. They have a diploid set of chromosomes. May survive diploid depending on age or tissue from which the cell culture was obtained. If the embryo is up to 80 days. For an adult – no more than 25 transplants. There are no more than 5 old ones. Transplanted ones are mutated cells that are cancerous. They last an infinite number of times. These are transformed cells of a cancer tumor. Hela is the most famous continuous cell culture since 1956. This culture is present in all laboratories in the world. It is adapted to many pathogens. First-born animals have a number of advantages: they do not die; higher growth rate; they are all genetically homogeneous. In laboptoria they are maintained by reseeding from one vessel to another.

59. CPD. This is a method for indicating the virus in cell culture. CPD refers to any changes in cells in a cell culture under the influence of a virus reproducing in them. I use low magnification when looking at the top layer of the mattress. Compare infected cells with uninfected ones. Differences may extend throughout the entire monolayer or only in patches. They are valued in kristas or points. So, if the entire monopoly of the CPU has undergone a change, it is estimated at 4 crosses; if ¾ - by 3 cr; ½ by 2 cr; ¼ - for 1 cross. The forms of CPD depend on the biological properties of the virus, the type of cells, the dose of infection, cultivation conditions, etc. Some viruses exhibit CPD after 2–3 days, others after 1–2. 3 forms of CPU: fragmentation– destruction of cells into separate fragments, which are separated from the glass and pass into the cultural fluid. Rounding– cells lose the ability to attach to glass, they take a spherical shape, separate and float freely where they die. Symplast formation– dissolution of cell membranes, as a result of which the cytoplasms of neighboring cells merge, forming a single whole in which the cell nuclei are located. Such formations are called symplasts - giant polyphagous cells. It is necessary to carry out at least 3 blind passages in order to judge the presence of the virus in the test material. Hemadsoption is the connection of red blood cells with the surface of virus-infected cells.

51. Calculation of virus titer according to Reed and Mench. Titration of the virus with a statistically assessed effect with calculation of the titer by read and menu. For this titration method, any biological model can be used, but this model must be sensitive to the virus being titrated (cell cultures, embryos, laboratory animals). According to the infectious effect of infected biological models, they are divided into the following: clinically recognized; according to pathomorphological changes; upon the death of the model; by the accumulation of hemagglutinin. The results of work depend on the dose of the virus. It has been established that the dose of the virus that causes 50 percent of the infectious effect is the least susceptible to fluctuations and is the most determinable of all possible doses. The titer is expressed in effective 50 percent doses. This is an ED of 50. Depending on the biological model used and the effect obtained, the 50 percent dose can be expressed in the following units: LD 50 – ID 50 ELD 50 EID 50 TsPD 50– this is a 50 percent cytopathogenic dose determined in cell cultures by CPD. If in infected systems we do not observe 50 percent of the effect that ID 50, then the titer is calculated by read and menu: lg LD 50 = lg ECD - (% years of ECD - 50%) / (% years of ECD - % years of ECD) ALL THIS MULTIPLIED BY lg multiplicity times

36. Rules and operating hours in a virology laboratory. All students are instructed and trained in safe tr. Entry into the production premises by unauthorized persons, as well as entry without a robe and replacement shoes, is prohibited. It is forbidden to go outside the lab wearing a robe and cap. Smoking, eating in the lab and storing food. All material entering the laboratory must be considered infected. At the end of the work, the workplace is put in order and thoroughly de-identified. Labeling of utensils containing infectious material. Hands wearing gloves are washed in a jar with a 5 percent chloramide solution, then the gloves are removed and disinfected for a second time, disinfected and washed. ra The laboratory's virologist work is built on three main principles: prevent infection of employees or people working with virus-containing materials. Prevent contamination of the material (tools, utensils are sterile) cleaning the premises with a disinfectant solution + ultraviolet lamps. Prevent the virus from being carried outside the laboratory (with air, utensils, solid and liquid material). Pipettes and glasses must be disposed of in the sterilizer. Test tubes with viruses, tissues - into an autoclave. Do not open the centrifuge until it stops. You only need to remove air from the syringe using a cotton swab with 75 percent alcohol. It is prohibited to ventilate the room using a ventilation system with a filter.

37. Safety precautions with virus-containing material. Prevent the dispersion of viruses in the external environment. Prevent contamination (contamination) of virus-containing material with foreign microflora. Ensure personal safety. To comply with these requirements, the following work rules are necessary: ​​be attentive and neat; be only in a robe and change in the wardrobe; work only with buttoned cuffs, a cap and a gauze mask; strictly maintain cleanliness and order in the laboratory; there should be no foreign objects on the desktop; Smoking and eating are prohibited. Use sterile instruments and utensils. Work with vessels near the burner flame. Do not put your fingers in your mouth. Used devices in the sterilizer. Collect used pipettes in a vessel with a disinfectant solution. Collect solid or liquid waste (cotton wool) in special containers for subsequent disinfection. Do not pour waste into sinks or toilets.

33. The mechanism of the antiviral action of interferon. Interferon does not have a direct effect on the virus. It affects only the cell by activating the synthesis of certain cellular enzymes. In particular, the enzyme protein kinase and 2,5 oligoasynthetase. Information about the synthesis of these enzymes is also located in certain regions of the cell’s genes and is also in a repressive state. Under air interf there is derepression of the genes responsible for the synthesis of protein kinase and 2,5 iligoAs synthetase. And their synthesis increases sharply. 1) under the air of protein kinase, the initiating factor is phosphorated, which ensures binding of the viral messenger RNA to the ribosome. Thus, the viral messenger RNA cannot contact the ribosomal apparatus of the cell, i.e., the beginning of translation. And ultimately, the synthesis of viral proteins and enzymes becomes impossible. 2) under the influence of interferon, the synthesis of 2,5 oligoAsynthetase is activated, which catalyzes the synthesis of 2,5 oligoadenylic acid in the cell. This acid switches the action of cellular nucleases to destroy viral messenger RNAs. Thus, under the influence of interferon, the following occurs: blocking the translation of viral messenger RNA; destruction of viral messenger RNAs. Inhibitory effect of interferon on cell reproduction: interferon in concentrations from 0 to 1000 units per ml suppresses the reproduction of a wide variety of cells in any tissue. Interferon regulates the growth of many types of cells including primary cell cultures and tumor cells. It is based on the suppression of the synthesis of certain cellular proteins and the synthesis of new proteins by interferon. Inter increases the killer activity of T-lymphocytes. In large doses, they inhibit antibody formation. Small doses, on the contrary, stimulate antibody formation. Krilling – cells treated with small doses of interf produce more interf than untreated cells. Too large doses - the opposite process.

35. Types of interaction between virus and cell. Productive and abortive. Productive is divided into lytic and latent. Productive: This is a type of interaction in which a new generation of virion is formed in the cell. If the cell quickly dies after acquiring a new virion, then this is a productive lytic pathway of interaction between the virus and the cell. If the cell in which the virus grows for a long time retains its viability (by budding), then this is a product of a latent type of interaction. Abortive: This type is mutual when the reproduction of virions stops at any stage, virion does not develop. As a result of the interaction of the virus with the cell, the following changes can occur in the cell: cell degeneration– cells first transform into an irregular shape, then become rounded, granularity appears in the cytoplasm, then nuclear fragmentation then cell death. Such changes are called CPP. In crosses: 4 crosses – 100% efficiency. Formation of symplasts– multinucleated cells. Formation of inclusion bodies– intranuclear and plasmatic mb, RNA or DNA containing. Cell transformation– oncogenic viruses (RNA retroviruses). Reproduction of oncogenic virs in a cell is not accompanied by CPD. The cell constantly produces a virus. Interferon synthesis.

4. Resistance of viruses to physicochemical factors. The resistance of animal viruses has been relatively well studied when exposed to external factors: temperature, radiation, ultraviolet, ultrasound, pH, formaldehyde, phenol, etc. To protect against these influences, virions have a protein shell. The different structure and chemical composition of protein shells determines the varying stability of viruses. Depending on these features, the same factor can destroy some virions completely and not others. For example, organic solvents: those virions in the shells of which there are no lipids are resistant to these substances, and those containing lipids are quickly destroyed. Inactivation of viruses means the complete or partial loss of their biological activity, which occurs as a result of the actions of physical and chemical factors. When the viral nucleic acid and protein change, complete inactivation occurs, i.e. loss of all biological properties of the virus - it loses only its infectious properties and retains its immunogenicity. The nature and extent of agents of a chemical and physical nature acting on the virus depends on the nature of the inactivating factor, on the dose, for a long time, on the type of virus. When the virus is inactivated, either the cleavage of the shell proteins can occur, followed by its disintegration into separate units, or the compaction of the proteins while maintaining the overall structure of the shell. Cleavage is observed under the action of an acidic and alkaline environment with prolonged and low heating. Coagulation and compaction occur when exposed to formaldehyde, high temperature or phenol. Depends on concentration and duration. Thus, in some cases, coagulation of proteins is accompanied by the destruction of nuclear acids and the virus experiences an irreversible loss of infectivity. In other cases, the virus's ability to reproduce is preserved. Preserved with glycerin.

60. PCR. The principle of the method: a gene specific to a given virus is identified - a section of a DNA molecule that carries information for the synthesis of one protein. This gene is then identified in the test material using PCR. This reaction allows the formation of additional copies of the gene - amplification of a section of DNA in a test tube. Depending on the purpose of the study, the species or genus of mo can be identified. The essence of PCR: the DNA molecule is heated to 90-94 degrees. Which leads to the destruction of hydrogen bonds between the nitrogenous bases of the double helix and then cooled to 52 g in the presence of the enzyme DNA polymyrase. A subsequent increase in the rate leads to the synthesis of a new DNA molecule - a complementary template. This procedure is repeated many times, resulting in larger fragments. Indication is carried out using electrophoresis or a labeled DNA probe. Main components: DNA polymyrase is thermostable; oligonucleotide of 20 nucleotides; triphosphates; amplifier, glassware and reagents for electrophoresis in agarose gel. Setup: obtaining a DNA sample. To do this, the material under study is suspended in buffer or distilled water. Add sodium OH and hold for 7 minutes. The mixture is neutralized. The lysate is centifuged for 10 minutes to sediment large particles. The supernatant liquid is used for PCR. PCR is the amplification of a given gene of a DNA fragment. Then it is melted in a thermal cycler for 3 hours. Indication of amplification - the sample is subjected to electrophoresis in an agarose gel to separate the DNA. After 30 minutes, the agarose is polymized in the apparatus and holes are formed in the agarose. 10 μl of the mixture is taken and mixed with 5 μl of dye. The mixture is added to the wells and electrophoresis is carried out for 40 minutes. The plate is removed and stained in a bromide solution for 10 minutes. The agarose is then placed on a transilluminator and the resulting band patterns are photographed. The bands revealed by ultraviolet radiation are DNA fragments.

49. Method of infection of cell cultures. Indication of viruses in cell cultures. Infection: for this purpose, tubes with a continuous cell monolayer are selected. The growth medium is drained and the cells are washed a couple of times with Hank's solution. 0.2 - 0.1 ml of virus material is added to each tube and distributed evenly over the entire layer of cells by shaking. In this form, the tubes are left for 1 to 2 hours at 22 or 37 degrees for the adsorption of the virus on the surface of the cells. Then the virus material is removed from the test tubes and the maintenance medium is poured into the test tube (1-2 ml). After isolation of the virus, the monolayer of cells is washed 2 times with Hank's solution and then the supporting medium is poured. Indication: according to CPD; RGAd; by plaque formation; intracellular inclusions; REEF; electron microscopy

54. RTGA. RGA. RTGA: essence– when the virus is mixed with a special serum, the virus loses its hemagglutinating properties. Goals– identification of isolated vir; detection of antibodies in the test serum and their titer. Components– for the direct serovariant: virus-containing material, specific serum, 1% suspension of red blood cells, saline solution for dilution. For retrospective - test serum, standard antigen in a certain dose of 4 GAE (virus dilution titer) 4 GAE - 1:32. Scheme– to each dilution of serum add an equal volume of standard antigen (virus) in a dose of 4 GAE. Contact 30 minutes at room temp. To each well with serum dilutions and a constant dose of virus in 4 ha, add an equal volume of red blood cell suspension. Contact 30-60 min at room temp. Accounting reactions are carried out in cristae. if it’s a plus, then there’s no agglutination; if it’s minutes, then it’s hemagglutination. The antibody titer in the test serum is the maximum dilution of the serum that completely delays the agglutination of red blood cells.RGA: essence: in the adsorption of the virus on the surface of the red blood cell, which leads to gluing. Purposes: indication; for virus titration in haen. Components: virus; 0.5 red blood cell suspension; saline solution for preparation. Post scheme: prepare a two-fold dilution of the virus; add an equal volume of 0.5% red blood cell suspension to each virus development; contact 30-60 minutes at room temperature. Accounting: in crists. 4 cristae – 100% agglutination. 3 crist - 75% . 1 cross – agglutination. 1 haen is the maximum dilution of the virus that can cause agglutination of 50% of red blood cells.

57. ELISA. The essence: when antigen + labeled serum binds, the enzyme decomposes the substrate. An antigen+conjugate complex is formed to form a colored reaction product, assessed under a light microscope or visually. Purpose: identification. Components: virus soda material, conjugate, substrate. Scheme of setting: the cell culitra is fixed with cooled acetone. They are dried and the conjugate is applied to them. Incubate for 1-2 hours at a temperature of 37 degrees in a humid chamber. Wash with saline solution, rinse with distilled water and dry. A few drops of the substrate solution are applied to it, incubated for 5-10 minutes, then washed in saline and rinsed with thin water. Accounting: in this case, that is, in the presence of antigen, after application of the conjugate, an antigen plus antibody complex labeled with an enzyme is formed. After application of the substrate, it is decomposed by the action of an enzyme, forming a colored product clearly visible in a light microscope.

56. RSK. The essence: the binding of a compliment to the antigen plus antibody complex. The absence of free compliment in this system is judged by the retention of hemolysin in the indicator system. Objectives: identification; detection of antibodies and their titer in the test blood serum. Components: 2 systems – 1 (virus-containing material; specific serum;) (test serum; standard antigen). 2) hemolytic system (indicator) - 2-3% suspension of sheep erythrocytes is an antigen; hemolysin (hemolytic serum) is an antibody. Antibodies correspond to the antigen. And a compliment for only 1 reaction: if the first, then there will be a delay in hemolysis when the compliment contacts the system under study. If on the second, then the red blood cells are lysed, there will be complete hemolysis. Setup scheme: the reaction is first carried out in the system under study, then the indicator system is added to the same test tube. Accounting: positive RSC – delayed hemolysis. Negative – complete hemolysis.

7. Viral proteins. Consist of amino acids. The composition of the viral protein depends on the order of alternation of amino acids; this order is determined by the genetic information contained in the viral genome. Viral proteins are divided into structural and non-structural. Structural proteins are part of mature virions. Non-structural b are not included in mature virions but are obligatory at certain stages of reproduction. StructuralNon-structural

8. Viral enzymes. They are protein in nature. They can be associated directly with the virion, but they are not associated - not structural. in DNA For RNA: RNA-dependent DNA polymyrase - it is not present in the cell, it is needed for “-” containing RNA and DNA, which in the vir family of retroviridae contains an enzyme that transduces the viral genome is called RNA-dependent DNA polymyrase. This enzyme has the names: revertase, reverse transcriptase. Enzymes involved in the formation of viral proteins: proteases, protein kenase.

52. RN.Essence: When the virus interacts with a specific serum, the virus loses its infectious properties, the ability to multiply in cells. Goals: identification of the isolated virus, detection of antibodies in blood serum and antibody titer. Components: virus-containing material, specific serum, biological model. If retrospective: test blood serum, standard antigen, biological model. General setup scheme: mix antigen and antibody, contact for 30-40 minutes, maximum 2 hours at a temperature of 37-38 degrees; a mixture of antigen plus antibody is used to infect a biological model; observation and accounting. Accounting: positive pH means alive, negative pH means dead.

53. RDP.Essence: the same antigen and antibody placed at the same distance from each other in an agar gel diffuse towards each other, forming a precipitate in the form of a white stripe at the meeting point. Goals: identification of the isolated virus, detection of antibodies in the test serum. Components: virus-containing material, specific serum, agar gel. For retrospective: test blood serum, standard antigen, agar gel. Staging scheme: agar coatings are prepared on a glass slide, wells are prepared, reaction components are added to the wells according to a certain scheme, the glass with the reaction is placed in a thermostat at 37-38 C. The reaction is recorded after 48 hours. A positive reaction is the formation of a white precipitation band.

55. RGAd, RTGAd. RGAd: Essence: in the adsorption of erythrocytes on the surface of virus-infected cells. Goals: virus indication. Components: cell culture infected with virus-containing material; red blood cell suspension . Staging scheme: pre-infect a single-layer cell culture with the test material. Cultures are drained into supporting culture medium. Wash with Hanks solution. A suspension of red blood cells is added. Contact 5-15 minutes at room temp. Accounting: carried out under a light microscope. Positive – red blood cells are adsorbed on the cells; negative – red blood cells float freely. RTGAd: gist: in the binding of specific antibodies to the surface of virus-infected cells, which leads to inhibition of adsorption on erythrocyte cells. Goals: identification of the isolated virus. Components: contaminated cell culture; specific serum; suspension of red blood cells. Staging scheme: a single-layer cell culture is pre-infected with a single-layer initial virus-containing material. Pour into a feeding medium and add 0.8 ml of specific serum. Contact 20-30 minutes. A suspension of red blood cells is added. Contact 5 – 15 minutes. Accounting: For control, they must install RGA. Accounting in experimental tubes: positive - red blood cells float freely, negative - red blood cells also float freely. Accounting in control tubes with positive – adsorption, negative – free floating.

6. Viral nucleic acids.+ RNA is a viral nucleic acid that also has the function of informative RNA. Information on the protein synthesizing system in RNA+ is immediately transferred to genomic RNA without transcription. -RNA containing viruses are viruses with single-stranded RNA that does not have the function of messenger RNA; in such viruses, the synthesis of messenger RNA (transcription) occurs on the template minus strands of genomic RNA using a virus-specific enzyme closely associated with gnome RNA, RNA-dependent RNA polymyrase. There are viruses containing both plus and minus RNA strands, these include adenoviruses and paramyxoviruses. Genomic information in double-stranded DNA is encoded on both strands. Nucleic acids are represented by polynucleotides consisting of individual nucleotides. Their quantity in nucleic acid varies. Each nucleotide consists of 3 subunits: a phosphoric acid residue, a carbohydrate, and a nitrogenous base.

9. Structure of viruses. Basic forms. Types of symmetry. Structure: DNA: usually double-stranded, gene information is encoded on both strands. Viral DNA can be arranged in a linear, circular fashion. Can be single-stranded. Viral RNAs: often single-stranded, less often double-stranded. Arranged linearly, in a circular manner, fragmented. As a rule, they consist of 11-12 fragments. Single-stranded vir RNA can be of two types: plus strands and minus stranded RNAs (negative genome). Types of symmetry: the location of protein subunits (caposmeres) determines the type of symmetry of the virion - helical, cubic, combined. Spiral This is a type of symmetry in which capsomers are located around the nucleic acid in a helical manner. Large viruses and some medium-sized viruses have this type of sim. Shape: rod-shaped, poly-shaped, spherical, oval. In rod-shaped viruses, the capsid consists of capsomers arranged around the nucleic acid in spiral turns of the same diameter, closely adjacent to each other. In spherical viruses, the capsomers are arranged in a spiral but of different diameters. Cubic type: Most small viruses and a significant proportion of medium-sized viruses have it. The shape of such viruses is spherical. The capsomeres of the capsid are located around the acid nuclei as around a regular isometric body. The protein shell of such viruses approaches the shape of an icosaider, a regular 20-sided face. Combined type of symmetry: consists of spiral and cubic. All phages and some complex viruses of the coxviridae family have it. They have a cubic outer shell and a spiral capsid shell. Phages have an icoseindrical head and a spiral process.

18. The main stages of the first phase of viral reproduction. This is the phase of infection of the cell, during this phase the virion must contact the cell, penetrate the cell and undress. First the stage of adsorption of virions on the cell surface can occur in two ways: physicochemical (non-specific); receptor (specific). The physicochemical pathway is determined by the interaction of surface electrostatic forces that arise between the positively charged groups of viral proteins and the negatively charged carboxins, sulfate, and phosphate groups of the cell wall. Receptor based on the specific interaction of the viral protein receptor with complementary receptors on the surface of the cell wall. Receptors of viruses and receptors of cells sensitive to a given virus have a complementary configuration (like a key to a lock). If the cell is not sensitive, then reabsorption will never occur. Second penetration – occurs in different ways for different viruses: with the help of viropexys; by fusing the shells. Viropexis– this path is similar to pinocytosis. First, at the site of adsorption on the cell surface, invagination of the cell wall of the membrane occurs, then the edges of the membrane close with the inside of the cell, the virion with all its membranes appears in the cell vacuole. By fusion- in this case, the areas of the viral shell and cell membrane that receive each other are melted under the action of virus-specific enzymes and only the viral nucleic acid appears in the cell, while the remnants of the virus are built into the cell membrane. Third stage– deprotenation – release from membranes – depends on the routes of entry of the virus into the cell. If deprotonization is not isolated as a separate stage by fusion of the membranes, it occurs simultaneously with the penetration of the virus. If penetration is through viropexis, then the release of the viral nucleic acid from the envelopes begins after the destruction of the proteins, lipids, and fats that make up the viral envelopes. All stages are temperature dependent.

20. Transcription. This is the rewriting of genetic information from viral nucleic acid to viral information RNA, newly synthesized according to the laws of the genetic code. (the virus must present the protein to the cell being synthesized and be converted into RNA). The end product of transcription is viral messenger RNA. Single-stranded + RNAs do not have transcripts, but their genomic viral RNA has the information vir RNA. In single-stranded -RNA, the genome cannot perform the function of messenger RNA and its RNA is transflexed using the virus-specific enzyme RNA-dependent RNA polymyrase. DRAWING!

21. Broadcast. This is the process of translating the genetic information contained in viral messenger RNAs into a specific amino acid sequence. A translation occurs when four bases embedded in the viral messenger RNA are converted into a code of 20 amino acids. The final product of translation is viral proteins. Protein synthesis occurs on cell ribosomes. Consists of 3 phases: initiation of translation and the beginning of translation; continuation; termination – end of broadcast. Initiation is based on the formation of a complex of components necessary for the start of translation, i.e., the initiation complex is also based on the recognition of the ribosome by the viral messenger RNA and its binding to certain areas called the cap. This is methylated guanine. After recognizing the cap, the ribosome slides down the messenger RNA molecule until it reaches the site where decoding of information begins.

5. Chemical composition of viruses. Viruses are made up of nucleic acids (DNA,RNA). Nucleic acids are represented by polynucleotides consisting of individual non-nucleotides. Each nucleotide consists of 3 subunits: a phosphoric acid residue, a carbohydrate, and a nitrogenous base. Viral proteins : Composed of amino acids. The composition of the viral protein depends on the order of alternation of amino acids; this order is determined by the genetic information contained in the viral genome. Viral proteins are divided into structural and non-structural. Structural proteins are part of mature virions. Non-structural b are not included in mature virions but are obligatory at certain stages of reproduction. Structural Depending on their location in the virion, viral proteins are divided into the following group: capsid proteins - in the capsid; supercapsid b – in supercapsid (mostly protein, there are also fats and carbohydrates); matrix proteins – proteins of the membrane layer; proteins of the viral core are represented by enzymes. Non-structural– depending on the functions they perform, they are divided into: regulator of viral genome expression; cell biosynthesis inhibitors; cell destruction inducers; viral protein precursors structural vir proteins; some viral enzymes are not part of mature virions. Lipids: are mainly included in part of the surercapsid shell of the virion in complex viruses. All of them are not encoded by the vir genome and are of cellular origin. They are represented by phospholipids and glycolipids. Carbohydrates: are part of the obol supercapsid, are not encoded by the viral genome and are of cellular origin, represented by glycoproteins and glycolipids . Viral enzymes: They are protein in nature. They can be associated directly with the virion, but they are not associated - not structural. The enzymes early polymyrases and early replicases take part in the stage of information change. They are classified as inhibitors of cellular biosynthesis. Enzymes transhibiting the viral genome: in DNA containing viruses - DNA-dependent RNA polymyrases are present in the cell; in some cases it is accessible to viruses, in others it is not. It can be of either cellular or viral origin. DNA-containing viruses that reproduce in the nucleus are of cellular origin. In the cytoplasm - viral origin - virus-specific.

For RNA: RNA-dependent DNA polymyrase - it is not present in the cell, it is needed for “-” containing RNA and DNA, which in the vir family of retroviridae contains an enzyme that transduces the viral genome is called RNA-dependent DNA polymyrase. This enzyme has the names: revertase, reverse transcriptase. Enzymes involved in the formation of viral proteins: proteases, protein kenase.

13. Bacteriophages. Bacteria virus. DNA and RNA bacteriophages are known. Most phage DNA is double-stranded. RNA phages are single-stranded. The phage nucleic acid is surrounded by a polyhedral capsid (head), to which in many phages an appendage (tail) is attached. The diameter of the heads is approximately 60-95 nm and the length of the processes is 250 nm with a thickness of 10-25 nm. The process serves as an attachment structure to the bacterium. The interaction between b and microbial cells is a complex biological process, the outcome of which depends on the properties of phages and is manifested by the lysis of bacterial cells. bacteriophages are used for diagnostics (anthrax); for the treatment of bacterial infections; for the prevention of inf (salmonellosis). DRAWING!

22. Replication of viral DNA.

23. Viral RNA replication. Single-stranded RNA with a negative genome:

25. Assembly of virions and their release from the cell.

: explosion, rupture, destruction of the cell in which mature virions (simple viruses) have formed, the cell dies. Complex structures come out by budding, that is, they come out through the cell wall and bud off. In this case, the cell does not die immediately, but when its reserves are used up

19. Second phase of reproduction.Eclipse stage: stage of information change. At this stage, the function of the cellular genome is suppressed due to the fact that nucleic acid blocks the virus and viral enzymes block the genetic apparatus of the cell and the cell’s synthesizing systems. This causes the cell to stop reproducing its own cellular components and switch to reproducing external components. Early replicases and early polymyrases are involved here. Transcription: This is the rewriting of genetic information from viral nucleic acid to viral information RNA, newly synthesized according to the laws of the genetic code. (the virus must present the protein to the cell being synthesized and be converted into RNA). The end product of transcription is viral messenger RNA. Single-stranded + RNAs do not have transcripts, but their genomic viral RNA has the information vir RNA. In single-stranded -RNA, the genome cannot perform the function of messenger RNA and its RNA is transflexed using the virus-specific enzyme RNA-dependent RNA polymyrase. DRAWING!Broadcast: This is the process of translating the genetic information contained in viral messenger RNAs into a specific amino acid sequence. A translation occurs when four bases embedded in the viral messenger RNA are converted into a code of 20 amino acids. The final product of translation is viral proteins. Protein synthesis occurs on cell ribosomes. Consists of 3 phases: initiation of translation and the beginning of translation; continuation; termination – end of broadcast. Initiation is based on the formation of a complex of components necessary for the start of translation, i.e., the initiation complex is also based on the recognition of the ribosome by the viral messenger RNA and its binding to certain areas called the cap. This is methylated guanine. After recognizing the cap, the ribosome slides down the messenger RNA molecule until it reaches the site where decoding of information begins. Viral DNA replication: Double stranded DNA replication: A double-stranded DNA molecule is first separated into 2 separate strands using cellular nuclease enzymes, then viral information DNA is formed on one of the viral DNA strands of which is the matrix. This occurs with the help of a virus-specific or cellular enzyme DNA-dependent RNA polymyrase. Then the viral infor RNA moves to the cell ribosome where translation occurs with the formation of viral proteins and enzymes, including the enzyme DNA polymyrase. Using DNA polymyrase, a second complementary strand of DNA is built from cell nucleotides. In this way, new double-stranded DNA molecules are synthesized. Replication of single-stranded DNA: DNA single strands have positive polarity. With the help of the virus-specific enzyme DNA-dependent DNA polymyrase, a complementary minus strand of DNA is formed on the viral single-stranded DNA matrix. A double-helix structure is synthesized, which is called the replicative form. Then, on the template, the minus strands of the replicative form form plus strands of single-stranded DNA by displacing the plus strands of DNA from the replicative form. Replication of viral RNA: Single-stranded RNA with a negative genome: Immediately after penetration into the cell, transcription occurs with the formation of viral inf RNA plus. The virus-specific enzyme RNA-dependent RNA polymyrase is involved in this. Vir inf RNA is then translated to form proteins and the enzyme RNA polymyrase. Subsequently, with the help of RNA polymyrase, daughter single-stranded minus RNA strands are formed on the matrix plus information RNA strands. Single-stranded RNA with a positive genome: After penetration into the cell, plus RNA immediately binds to ribosomes where it is translated to form proteins and enzymes, including the RNA replicase enzyme. Then, under the action of RNA replicase, a replicative form is formed. On the template, the minus RNA of the replicative form creates an image of plus RNA strands by displacing them from the replicative form. Replication of double-stranded viral RNA: the synthesis of messenger viral RNAs occurs on a double-stranded RNA template using the enzyme RNA-dependent RNA polymyrase. Transcription on an RNA strand template, each fragment is transcribed separately. Then they are translated to form proteins and the enzyme RNA polymyrase with the help of this enzyme on the plus strands of messenger RNA to form complementary minus strands of RNA, i.e. double-stranded RNA. Assembly of virions and their release from the cell: 2 strategies for assembly, maturation and exit from an infected cell: implementation of assembly and maturation within cells; combination of the last stage of virion assembly with exit from the infected cell.

Assembly is carried out by simple aggregation, i.e. the combination of a vir protein with a nucleic acid occurs under the influence of physicochemical factors, i.e. self-assembly occurs. It is based on unification and specific non-protein and protein-nucleic acid recognition. A nucleocapsid is formed. For simple viruses, this is where the self-assembly process ends. In complex viruses, the process of self-assembly is carried out differently. Part of the protein goes to the formation of the nucleocapsid, which is formed like in simple viruses, and part of the proteins moves to the cell membrane. The formed nucleocapsid subsequently moves there. And the formation of a supercapsid shell occurs when the virion leaves the cell, i.e. the nucleocapsid is covered on top with proteins that have moved to the cell membrane and upon exiting fats and carbohydrates from the cell are built into this outer shell . There are two ways to exit: explosion, rupture, destruction of the cell in which mature virions (simple viruses) have formed, the cell dies. Complex structures come out by budding, that is, they come out through the cell wall and bud off. In this case, the cell does not die immediately, but when its reserves are used up.

62-63. Sheep and goat pox(genus Caprippoxvirus). Contagious ecthyma of sheep and goats(genus Parapoxvirus).Family: poxviridae. DNA containing. Features of reproduction: the genome of the virus is very large. Even in infected cells, replication is completed after 6 hours. Penetration occurs by fusion of the viral and cellular membranes. After penetration, double-stranded DNA is split and replication begins on both DNA strands at once. Moreover, the synthesis of viral components occurs in the cytoplasm of cells. Virion assembly in the cytoplasm. Exit by budding.

2. The role of viruses in infectious pathology of animals. Currently, viral diseases are of great importance in the infectious pathology of animals, humans and plants. Their role increases with the reduction and elimination of bacterial, mycotic and protozoal diseases. Viral diseases account for approximately 80 percent in medicine and 50 percent in veterinary medicine. There are over 500 known diseases caused by viruses. Viral origin has been established in such particularly dangerous diseases as foot and mouth disease, rinderpest, swine fever, etc. Virology can be divided into general and specific. General studies the nature and origin of viruses, their classification, structure and chemical composition, genetics and selection, methods of diagnosis and prevention, the basics of antiviral immunity. Private vir studies the name and systematic position of specific pathogens, the structure, size, and stability of the virion, therapy, diagnostic methods, and prevention.

1. History of development. The first period begins from ancient times until 1892. During this period, virology as an independent science did not exist. Bacteriologists studied diseases of unknown etiology. The second period - the formation of virology as a science itself - covers 1892-1950. this period began with the discovery of the Russian botanist D.I. Ivanovsky (1898) on the filiability of the causative agent of tobacco mosaic disease. Ivanovsky, studying the etiology of tobacco disease, established that this disease is caused by a special tiny microorganism that passes through bacterial filters. It is invisible under a light microscope. Does not grow on artificial growth media. Subsequently, similar MOs were isolated from other plants, as well as from animals and humans. They were united into an independent group - ultraviruses. In the 1930s, chicken embryos began to be used in virological practice. In 1956, Stanley succeeded in dividing the virus into its main components - protein and nucleic acid. In the late 40s of the 20th century, the creation of electronic micros Rudenberg. And the light one was created by Leeuwenhoek. Soviet scientists who contributed to virology: Zhdanov, Likhachev, Syurin.

48. Method for obtaining single-layer primary trypsinized cell cultures. Single-layer cells are needed to indicate the virus. It is obtained from tissues or organs by treating them with trypsin. Single-layered ones are divided into 4 groups: primary trypsinized; subcultures; diploid or semi-grafted; grafted. The tissue is crushed and dispersed with the enzyme trypsin. Then trypsin is removed by centrifugation and a certain volume of liquid nutrient medium is added to the resulting sediment. The cells are grown in a single layer—a monolayer—on the inner surface of the glass. There is a constant need for organs from healthy animals. AND embryos 9-112 days old are used. Ovoscopy. Shell processing. Using scissors, cut off the shell above the border of the puga. The embryo is sterilely removed. Wash with Hank's solution. A skin-muscular sac is prepared. Wash with Hank's solution. The fabric is shredded with scissors. Transfer to a trypsinization flask. The flask is placed on a magnetic stirrer for 15 minutes. The suspension is cooled in a vessel with ice. Filter into a flask receiver. The cell suspension is centrifuged for 10-15 minutes. Trypsin is removed. A combined mass is prepared from the cell sediment, poured into 1 ml tubes and the cells are cultured.

47. Basic solutions and nutrient media.By origin There are natural feeding media and artificial feeding media. Naturals - from biologically active ones: embryonic, allantoic vitality plus the addition of balanced salt solutions. Artificial - prepared from individual components. Most often, universal feeding media are used, or there may be special ones. Universal is medium 199 and Igla medium. The composition of art media should include amino acids, vitamins, enzymes and balanced salt solutions, sometimes an indicator (phenol red). The essence of the indicator is to detect the virus by changing color. During the life of the cell, the pH changes to the acidic side. In an acidic environment, the color of the indicator changes from crimson red to yellow. Normal blood serum is sometimes added to the nutritional medium in a volume of 100 percent of the volume of the nutritional medium. Blood serum is called growth factor. It is added for cell proliferation, only in growth media . By purpose of use: growth media - serum included; supporting – no serum. Balanced salt solutions: all of them are derivatives of saline solution. Used as a basis for preparing pit media and for all manipulations with cell cultures (to wash something). These are Hanks and Earle solutions. Dispersing solutions: for separating cells from others and cells from glass. Solutions of pepsin, trypsin. From glass - versine solutions. The versine solution binds calcium cations.

50. Virus titer. Virus titer is the amount of virus, i.e. the dose per unit volume of virus-containing material. 3 titration methods: 1 method: titration of the virus according to the infectious effect of the virus with a statistically assessed effect. According to the method of read and menchu ​​or according to Kerber. The titer is expressed in 50% doses. This is ED50. For this titration method, any biological model can be used, but this model must be sensitive to the virus being titrated (cell cultures, embryos, laboratory animals). According to the infectious effect of infected biological models, they are divided into the following: clinically recognized; according to pathomorphological changes; upon the death of the model; by the accumulation of hemagglutinin. The results of work depend on the dose of the virus. It has been established that the dose of the virus that causes 50 percent of the infectious effect is the least susceptible to fluctuations and is the most determinable of all possible doses. The titer is expressed in effective 50 percent doses. This is an ED of 50. Depending on the biological model used and the effect obtained, the 50 percent dose can be expressed in the following units: LD 50 – This is 50 percent of the lethal dose received for the lab, which is alive in terms of the lethal effect. ID 50– this is a 50 percent infectious dose determined for the lab to be alive based on clinical signs or pathomorphological changes. ELD 50– this is a 50 percent embryonic lethal dose determined on chicken embryos by year of outcome. EID 50– this is a 50 percent embryonic infectious dose determined in chicken embryos by pathomorphological changes and the accumulation of hemagglutinin. TsPD 50– this is a 50 percent cytopathogenic dose determined in cell cultures by CPD. If in infected systems we do not observe 50 percent of the effect that ID 50, then the titer is calculated by read and menu: lg LD 50 = lg ECD - (% years of ECD - 50%) / (% years of ECD - % years of ECD) ALL THIS MULTIPLIED BY lg dilution factor. Method 2 : on the infectious effect of the virus with an assessment of the single effect. With the plaque formation method in cell culture, there is a single effect. Expressed in smallpox-forming units or plaque-forming units PFU. Prepare a 10-fold dilution of the virus; select a sensitive biological model; In each dilution of the virus, at least 4 embryos are infected. The titer is calculated using the formula T=a divided by V*n. a - the average number of pockmarks or plaques. V is the volume of material content = 0.2. n is the degree of dilution of the virus. Method 3: according to the hemagglutinating effect of the virus in GAEN. They put RGA.

38.Principles of laboratory diagnosis of viral infections.

Laboratory tests play an important role in establishing the diagnosis of infectious diseases, prescribing etiotropic therapy, and monitoring the effectiveness of treatment. The process of specific laboratory diagnostics is based on identifying the pathogen and the response of the human body during the infectious process. It consists of three stages: collecting material, transporting it (spur No. 39), and studying it in the laboratory: 1) The virological method includes two main stages: isolation of viruses and their identification. To isolate viruses, cell cultures, chicken embryos, and sometimes laboratory animals are used. The presence of the virus in infected cultures is determined by the development of specific cell degeneration, i.e. cytopathogenic effect, detection of intracellular inclusions, as well as based on the detection of a specific antigen by immunofluorescence, positive hemadsorption and hemagglutination reactions. Viruses are identified using immunological methods: hemagglutination inhibition reaction, complement fixation, neutralization, gel precipitation, immunofluorescence. 2) Serological reactions; 3) Immunological method (bioassays); 4) ELISA and PCR. After receiving the examination results and taking into account epidemiological and clinical data, a final diagnosis is established.

39. Taking, preparing and forwarding patent. Material for virologist. Research.

Collection, transportation and examination of Pat. material is regulated by veterinary legislation. When taking, the tropism of the virus is taken into account - the preferred localization of the virus in a given disease, and pathogenesis. Time from taking a stalemate. material until the end of its research - 2-4 hours. If you need more time, preserve it (chemical methods - 50% glycerol solution, physical - freezing), but not for luminists. microscopy. Transportation to special containers with accompanying document and courier. Preparation involves extracting the virus from the cells. Liquid material – filtration and centrifugation. To cleanse bacteria - bacterial. filters and antibiotics (500-2000 units per 1 ml), for fungi - fungicides (25 units per 1 ml), keep for 30-40 minutes, inoculate with pitata. environment (aerobes – MPA, MPB, MPZh, anaerobes – Kitta-Tarozzi, fungi – Chapeka, Saburo). Dense patent material: 1) take patent material 1-1.5 g; 2) chop with scissors; 3) wash with sterile. glass or sand in a mortar; 4) 10% suspension with Hanks solution; 5) freeze and thaw 2 times; 6)filtration through a gauze filter; 7) centrifugation (3000 rpm, 15 min); 8) supernatant liquid - virus-containing material, it is tested for bacteria (nutrient media), antibiotics and fungicides are added.

40. Microscopic method of research in virology.

1. Light microscopy: 1) for detection of smallpox virus (Morozov’s silvering method); 2) To detect inclusion bodies (this is an accumulation of virions or from parts or products of the cell’s reaction to the virus; they could be intranuclear and cytoplasmic); 3) detection of virus CPD (rounding, fragmentation, death); 4) detection of symplasts; 5)work with C/C; 6) assessment by serologist. reactions (ELISA, RGAd, RTGAd). 2. Luminescent microscopy: the essence is that when irradiated with UV rays, atoms are excited, then go into the initial state with the release of energy in the form of light radiation, its intensity is assessed in crosses (emerald-green = ++++; green = ++ +; green-yellow = ++; yellow = +; no glow = –). Before irradiation, the drug is painted with fluorochromes (FITC, acredin orange, yellow, rhodamine). This is simple fluorochrome plating. Complex - MFA. The essence of MFA is specific. interaction of the antibody with fluorochrome-labeled serum (conjugate). 3.Electron microscopy: 1) detection of any virus; 2) study of its size, shape, structure, type of symmetry, reproduction; 3) study of the interaction of the virus with the cell.

In the virology laboratory, work is carried out to isolate virus strains, their identification and cultivation, and various scientific studies are carried out. When working with viruses, you must first of all:

1. Prevent contamination of virus strains with foreign microflora;

2. Ensure the safety of working personnel from possible infection by viruses;

3. Ensure the safety of the surrounding population from infection with viral infections through wastewater, corpses of experimental animals, etc.

When studying materials obtained from patients with viral infections, for the purpose of laboratory diagnosis of these diseases, various methods are used:

· Methods of electron and, to a lesser extent, light microscopy;

· Methods for isolating and cultivating viruses in cell cultures;

· Methods for isolating and cultivating viruses in developing chicken embryos and in the body of sensitive experimental animals;

· Identification of viruses by their hemagglutinating ability;

· Various serological research methods: traditional and express methods;

· Molecular genetic research methods - molecular hybridization and polymerase chain reaction.

1.1.2. Materials studied for viral infections

When taking infectious material from people and animals, it is necessary to take into account the tropism of viruses for certain tissues and organs, the route of release of the virus into the external environment and the characteristics of the pathogenesis of a particular viral infection.

There are pneumotropic, enterotropic, hepatotropic, lymphotropic, neurotropic and dermotropic viruses. Depending on the tropism, various materials are subjected to research. For example, they examine mucus from the throat, sputum, etc., if the virus is pneumotropic; bowel movements - with enterotropic viruses; liquid from vesicles or pustules, crusts - if the virus is dermotropic, etc.

1.1.3. Processing of virus-containing material

Infectious materials, taken taking into account the tropism of viruses and in compliance with asepsis, are placed in sterile containers, carefully sealed and sent to the laboratory, placed in a thermos with ice.

It is recommended to examine the material as soon as possible, since viruses are quickly inactivated. The preservation of the virus is facilitated by placing the test material (in a 50% glycerin solution) in a refrigerator at a temperature not exceeding 5 o C. But the most reliable method is frozen storage at a temperature of -45 o C and below; under such conditions the virus can remain viable for a long time.

Processing of dense material containing viruses begins with grinding it in a mortar or grinding it in special devices - homogenizers. Then a 10% suspension is prepared in saline solution, which is centrifuged at 2000-3000 rpm for 15-30 minutes to sediment large particles. The viruses remain in the supernatant, which is subjected to further study.

The liquid virus-containing material is directly centrifuged and the supernatant is also obtained.

If there is doubt about the bacteriological sterility of the test virus-containing supernatant, antibiotics are added to it to destroy foreign microorganisms. Antibiotics do not affect viruses, and they remain viable.

1.1.4.Microscopic research methods in virology

- Electron microscopy

Electronscopic preparations are prepared from purified and concentrated virus-containing suspensions or ultrathin sections of tissue infected with viruses. Viral objects are applied to special substrate films placed on support meshes. Substrate films must be very thin (no more than 30 nm thick), transparent and sufficiently strong, for example, colloidal carbon. The films are applied to supporting copper meshes (2-3 mm in diameter) with numerous holes. The drugs are then processed in various ways.

Metal spraying methods used to obtain contrast agents. Vapors of heavy metals (gold, platinum, uranium, etc.), formed in a special device under vacuum and high temperature conditions, are directed at an acute angle onto the virus-containing drug. Viruses are coated with a thin layer of metal.

Negative contrast method is based on the fact that when the drug is treated with certain salts of heavy metals, for example, a 1-2% solution of phosphotungstic acid, a denser layer is created that does not allow electrons to pass through, and in which more electron-transparent objects under study are clearly visible.

Ultrathin sectioning combined with negative contrast is the best for studying the fine structure of virions and studying the stages of interaction of viruses with the cell, but at the same time it is the most complex. The examined pieces of infected tissue or other virus-containing material are fixed in a special fixative (for example, osmium). Dehydrate by sequential placement in alcohols of increasing strength. The samples are filled with special plastic, after polymerization of which solid transparent blocks are formed. Ultrathin sections with a thickness of 10-20 nm are prepared from the blocks on a special microtome. The resulting sections are contrasted by placing them in a solution of phosphotungstic acid.

The preparations prepared by the methods described above are studied in a transmission electron microscope, the resolution of which reaches 0.2-0.3 nm. The image of the preparation is observed on the fluorescent screen of an electron microscope and special photographic plates are photographed from which prints are obtained. Received magnifications: ×100000-×400000.

Scanning electron microscopy is carried out using a scanning electron microscope, in which a thin beam of electrons quickly moves across the object under study, that is, scans its surface. As a result, radiation of secondary electrons arises, which, passing through a cathode ray tube, is converted into a three-dimensional image of an object on a fluorescent screen.

Scanning microscopy makes it possible to obtain a three-dimensional image of virions (the preparation is first sprayed with metals), to distinguish the details of the structure of their surface, but does not reveal their internal structure. The resolution of a scanning microscope is 7-20 nm.

- Light microscopy

In a light microscope you can see large viruses, the sizes of which are within the resolution of the microscope - at least 0.2 microns. As well as intracellular inclusions in tissues affected by the virus.

Large viruses, for example, poxviruses, and inclusions are detected using special staining methods, in phase contrast, in a dark field of view; Fluorescent microscopy is also used.

Large viruses are identified by Morozov staining (silvering). To identify intracellular inclusions, histological sections from affected tissues, smears or prints are prepared. Usually the preparations are stained according to Romanovsky-Giemsa, sometimes by other methods. The detection of Babes-Negri inclusions in nerve cells of the brain during rabies is of greatest practical importance. For this purpose, the preparations are stained according to Mann.

Luminescence microscopy. Preparations prepared from materials containing large viruses, intracellular inclusions, and accumulations of viral antigens are stained with solutions of fluorochrome dyes. With fluorescent microscopy in UV light, acridine orange-stained accumulations of RNA genomic viruses and the inclusions they form are visible as luminous red granules against the background of pale green cell cytoplasm; DNA genomic viruses give off an emerald green glow.

Immunofluorescent method is based on the combination of viruses, intracellular inclusions, and accumulations of viral antigens with specific antiviral antibodies labeled with fluorochrome dyes. The resulting complexes glow under fluorescence microscopy.