Feb 16, 2022

The Structure of SARS-CoV-2 is the Smoking Gun for its Lab Origin – All That is Known About the COVID-19 Origin with All Relevant References

History of the Coronaviruses and the Wuhan Institute of Virology

Coronaviruses (CoVs) have been broadly studied in China since 2003, and numerous new strains have been reported. Wuhan coronavirus scientist Dr. Shi Zheng-Li (called “Bat Woman“ by media) co-authored many of the publications (10). Shi, employed in Wuhan Institute of Virology (WIV), also worked on enhancing the ability of bat viruses to attack humans, supposedly to “examine the emergence  potential of  circulating  bat CoVs”, which  means their ability to “jump over  to  humans”.  Inter species infections had already been artificially achieved before; in 2000, for example, the spike glycoprotein of the coronavirus mouse hepatitis virus had been artificially modified, so that the resulting chimeric virus acquired the ability to infect cat cells and lost the ability to infect mouse cells (30). Shi, however, tried to modify an animal virus to be able to infect human cells, which modification is called “gain-of-function”, and such experiments should be very strictly controlled. In 2015, Shi and coworkers modified a harmless bat coronavirus to be capable of infecting human cells (1); the work was strongly criticized as a dangerous experiment without any clear merit (11).

In 2017, an article in the prestigious journal Nature warned about the WIV; the labs were built to withstand a magnitude 7 earthquake, but the Western scientists worried about possible escapes of the viruses even without any earthquake, in view of known repeated escapes from the high-level containment facilities in China (2). In 2018, the U.S. Embassy in Beijing warned the U.S. government about safety and management weaknesses at the WIV labs, mentioning a risk of a new SARS-like pandemic (12 and 3). Strangely, in September 2019, the Global Preparedness Monitoring Board of the WHO, consisting mostly of Western scientists including Anthony Fauci, published an “Annual report on global prep for health emergencies”, named “World at Risk”, in which they warned about having to be prepared for the worst in a possible rapidly spreading, lethal respiratory pathogen pandemic (34). It turned out later that the corona epidemic started much earlier than in December 2019 when the Chinese authorities admitted it, probably in August 2019 or earlier (14, 15, 16), so that the above “prophetical” Annual WHO report had probably been inspired by the first epidemic indications.

Epidemic of COVID-19 Caused by a New Virus and Quick Identification of the Virus by the WIV

On December 31, 2019, the Chinese authorities reported to the WHO an “epidemic of pneumonia of unknown origin” in the city of Wuhan (23, 24, 25), while locating the origin of the epidemic to the Wuhan meat market around December 12. Despite the proclaimed novelty of the infection and its recent origin, the Chinese scientists succeeded to fully characterize the infectious agent as early as on January 7, 2020, namely merely seven days after reporting the epidemic, in an article forwarded to the journal Nature. According to the article, a patient with an unknown disease who worked at the local meat market was hospitalized on December 26, 2019; a specialized facility began looking for an unknown infectious factor responsible for the lung finding; extensive RNA sequencing of the patient's bronchial sample (over 56 million sequences read) resulted in identifying a new virus, including its entire structure and even its ability to bind to the ACE2 receptor through which it penetrates human cells; the evolutionary relationship of the new coronavirus to other viruses was also determined, and it was found that it is closely related to a bat virus recently found in China (17).

Respiratory problems can be caused by a wide range of viruses, bacteria, and fungi; in this case, the unknown infection factor was accurately determined at a record rate. According to the article, the researchers began looking for an unknown infectious factor around January 1, and within six days they were able to identify and fully characterize the new virus, allegedly by studying only one patient. However, the authors of the article, researchers at the University of Shanghai, were surely in contact with researchers of the WIV, who in parallel related to “the epidemic that started in Wuhan on December 12”, and who examined samples from seven patients (six of whom worked in the local meat market), reached similar conclusions as the first group and sent their results to Nature magazine on January 20 (18). The second article further announced the existence of a bat virus, called CoV RaTG13, which surprisingly had very high genome homology (sometimes called similarity or identity), namely 96% of RNA sequences, with the newly discovered virus, now called SARS-CoV-2 (shortly CoV-2), which similarity should wishfully confirm the alleged animal origin of CoV-2. The leader of the Wuhan Group was Shi Zheng-Li, and China probably wished that the virus, which caused the global world epidemic, be discovered by someone else than "the Bat Woman", but the journal Nature decided to publish both articles simultaneously on February 3.  

From the beginning, China disinformed about the epidemic. As mentioned above, the epidemic had been already circulating in Wuhan in August 2019. The first WHO Situation Report of January 2020 (25) already mentioned the spread of the disease in Beijing and Shanghai, which would indicate a nationwide epidemic by December, and the Report even mentioned spreading of the disease (now called COVID 19) to other countries. However, Dr. Shi, in an interview to Scientific American in February 2020 (5), stated that her boss had told her only on 30 December 2019 that a new coronavirus had been detected, allegedly mentioning two patients and asking her to examine the issue. So that the authors of the first article hardly searched for an unknown factor in one patient on January 1, when coronavirus had already been identified in more patients at that time; likewise, the authors of the second article had only confirmed in the seven received samples what had already been known to the Bat Woman’s boss. Even the CEO of the WIV, Ms. Wang Yanyi, contradicted all the known facts when she said in a later interview in 2020: “Our institute first received the clinical sample of the unknown pulmonary disease on December 30 of the last year. We had not had any knowledge before that, nor had we ever met, researched, or kept the virus. In fact, like everybody else, we had not even known about the existence of the virus” (42).

Hypothesis of a Natural Origin of the New Virus is Popular but Fails

Although the Chinese authorities even tried to accuse the United States of bringing the virus to China (21), their principal version for the virus origin comprised an animal source, from which SARS-CoV-2 had “naturally” evolved. The “natural origin” of the new virus was generally supported by the world mainstream media from the beginning, an example being condemnation of any attempts to look for an artificial origin in Guardian on February 20, 2020: “Experts fear false rumors could harm Chinese cooperation on coronavirus” (19). The scientific media joined the intimidating tone. For example, the Nature editors retroactively added a note to their article of 2017 (2) saying that the theory of the virus lab origin is unverified (the note was later removed). Lancet of March 7, 2020 stated: “We stand together to strongly condemn conspiracy theories suggesting that COVID-19 does not have a natural origin” (20). Said Lancet proclamation was drafted by Peter Daszak who, although stating no conflict of interests, had funded the coronavirus research at the Wuhan Institute of Virology (28) and, furthermore, is a co-author of the above infamous “gain-of-function” work, together with the “Bat Woman” (1). The ubiquitous Daszak is also a member of the WHO team investigating the origin of SARS-CoV-2.

The structural studies of the new virus in early 2020 showed that SARS-CoV-2 appeared to be optimized for binding to the human receptor ACE2, and even had newly acquired 12 nucleotides in its genome enabling it an easy entry to the human cells; however, after reporting said facts, the authors quickly emphasized that “It is improbable that SARS-CoV-2 emerged through laboratory manipulation” (35). The principal factors, considered in the virus origin, were zoonotic transfer probably via an intermediate animal host, followed by natural mutations (6, 7, 35, 36, 37, 38, 39). But some researchers, who are cited less broadly, considered also eventual artificial interventions, and they did ask more questions (13, 29, 26, 27, 22, 9) . Whatever the answer, this debate was not only legitimate but vital, as researcher Xiao Qiang of the University of California, Berkeley, said: "I don't think it's a conspiracy theory. This is a legitimate question that needs to be addressed and answered. Knowing exactly how this came about will help prevent it from happening again” (3, 12).

Very soon after the epidemic break, the Chinese scientists published genomes of animal viruses Bat-CoV RaTG13 and Pangolin-CoV 2020, implying that a zoonotic transfer from bats via, for example, pangolins was feasible, particularly in view of very high similarity of some of the proteins of the viruses (7), but the attempts to explain the natural path from an animal host to humans were not persuasive. The closest virus to SARS-CoV-2 published so far, RaTG13, has 96% RNA homology with CoV-2, which is too distant (corresponding to several decades of separate evolution) to explain a recent jump, notwithstanding the strange origin of this strain, miraculously provided by Dr. Shi Zheng-Li just several weeks after epidemic announcement by China, even thou the strain had allegedly been isolated in 2013. Despite the fact that Daszak and Shi had sampled more than 10,000 bats and 2000 other species, and found some 500 novel coronaviruses, according to a Science article (39), no virus was closer to SARS-CoV-2 than RaTG13, and the COVID-19 natural origin remains unsupported.

Hypothesis of a Laboratory Origin of the New Virus is Disliked but Prevails

Because the epidemic started close to the high security Wuhan Institute of Virology, an escape route from the laboratory is among the considered scenarios, also in view of the numerous such cases and the low level of security in the Chinese laboratories (2, 3). Incredible sloppiness of the work in the “highest security” Wuhan labs was indicated by the reports about experimental animals appearing on the local meat market as some Chinese researchers sell the laboratory animals to street vendors (4). Even Dr. Shi Zheng-Li herself inadvertently admitted, in her mentioned interview for Scientific American given in February and published in April 2020, that she had wondered whether the new virus could not originate from her laboratory (“If coronaviruses were the culprit, she remembers thinking, ‘Could they have come from our lab?) (5).

On February 20, 2020, soon after the corona epidemic had officially started, an important feature of the newly emerged virus was published: an insertion of 12-nucleotides into the RNA of SARS-CoV-2 within the S1/S2 border region, resulting in four extra amino acids in positions 681-684 of the spike protein, which binds to the human receptor ACE2, improves contagiousness of the virus, the extra residues being unique to this human virus and not being found in any other species (32). The inserted aminoacids PRRA create a cleavage site in the S1/S2 region for the human protease furin, facilitating entry into the target cells. Within three weeks, the Chinese scientists and their coworkers hurried to neutralize the unpleasant finding of the unique furin cleavage site in the CoV-2 genome, and they published a newly disclosed bat corona virus RmYN02 which had “natural insertions of multiple amino acids at the junction site(36), supposedly demonstrating that insertions like said new furin clevage site in CoV-2 arise easily and naturally; however, the allegedly found insertion was PAA, which is not a furin site, and moreover, it was later found that no insertions were in fact present in said region of RmYN02 at all (26). Attempts to explain the path from an animal host to human were failing.

The mainstream media have not yielded in their resistance toward the artificial origin of CoV-2. Even in February 2022, it was reported that “the complex evolutionary history of SARS-related coronaviruses is disentangled” (8), but at the same time it is admitted that a direct proximal ancestor (an animal virus with at least 99% homology with SARS-CoV-2) is still missing; the source of this news, by the way, is University of Glasgow, which hosts a Centre for Chinese research and Confucius Institute. On the other hand, scientific media slowly started to admit there were two legitimate hypotheses: natural origin versus lab origin. For example, Nature wrote about a new hypothesis of “multiple animal origins” in September 2021 as follows: “…new finding could be the dagger into the heart of the hypothesis that SARS-CoV-2 escaped from a lab, rather than originating from wildlife trade” (40). The article at least acknowledged that a lab escape was also a hypothesis and not a conspiracy theory.

Segreto and Deigin’s Review of Both Hypotheses: The Critical Insertion in the CoV-2 Spike Protein Could Hardly Occur Spontaneously and it Employs Codons Rare for Coronaviruses

A possible laboratory origin of the new virus was outlined in a well founded and detailed review of Segreto and Deigin in November 2020 (26). The authors, thoroughly supporting all their statements by relevant references, defined the two main SARS‐CoV‐2 features: (1) the presence of a furin cleavage site missing in other CoVs, and (2) a receptor binding domain (RBD) optimized to bind to human cells. The simultaneous acquisition of both unique features could hardly be natural or caused by serial passage, but they could be easily acquired by lab manipulation techniques such as site‐directed mutagenesis. Chinese researchers had been generating chimeric CoVs in WIV for years, and the authors carefully documented how, during the years, the researchers had disinformed the public regarding the dates when they had found the new strains and sequenced them, including the famous RaTG13.

Interestingly, the authors noted that two joint arginins in the acquired furin cleavage site are coded by two CGGCGG codons, which are rare for these viruses, and CGGCGG in the new insert is the only doubled instance of this codon in the whole 30,000 nucleotides genome of SARS‐CoV‐2. Moreover, the CGGCGG insert includes a FauI restriction site, which allows using restriction fragment length polymorphism (RFLP) techniques for cloning or screening for mutations.

The authors further mentioned the fact that RaTG13 and the pangolin CoV sequences, used in proving the hypothesis of natural origin of SARS‐CoV‐2, had been questioned as to the accuracy of their assembly data and had required further analyses to prove their correctness. Another grave issue was mentioned: modification and subsequent deletion of WIV's own viral database during 2020. The authors concluded that the leak of highly dangerous pathogens from laboratories was not a rare event, the most notable known being 1977 H1N1 lab escape from China that had caused a worldwide pandemic, and the most recent one being the November 2019 outbreak of brucellosis that had occurred in two research centers in Lanzhou, China, infecting over 100 students and staff members.

Sallard et al.’s Review of Both Hypotheses:  WIV was Caught Cheating About the Origin of Various Coronavirus Strains

In another review, Sallard et al. traced the origin of the virus in February 2021 and discussed the scenarios of a zoonotic emergence versus an accidental escape of a laboratory strain (22). Peculiarly, the authors found an exact match between RaTG13 and an RNA fragment of another bat which was published by Shi in 2016, which is strange, since RaTG13 was allegedly found in 2013 and published by Shi in 2020 – this added still more to the obscurity of RaTG13. Said RNA fragment, moreover, originated from a mine shaft in Yunnan Province following the death of 3 miners from an atypical pneumonia.

In any case, the authors believed that the SARS-CoV-2 genome is a “mosaic” genome composed of pieces of at least two preexisting CoVs. Further they noted that the natural origin could not be defended in the absence of a direct proximal ancestor, and alternative routes had to be necessarily considered. For example, SARS-CoV-2 might have been manufactured in a laboratory, or it may have resulted from a bat virus that became adapted to other species in laboratory animal models and then escaped from the laboratory. It may have also come from a viral strain that was cultured on human cells in a laboratory in order to study its infectious potential, and that was progressively “humanized” (adapted to humans) by selection of the viruses having the highest ability to spread in these conditions. 

Nicholas Wade’s Review of Both Hypotheses:  The Virus Was Adapted to Human Cells as Described in the Grants Assigned to Peter Daszak and Subcontracted to Dr. Shi Zheng-Li

Nicholas Wade, editor and author who had worked on the staff of Nature, Science, and the New York Times, summarized the known facts in May 2021, and showed that the origin of COVID-19 in one of Wuhan labs was hardly refutable (28). Wade noted that SARS-CoV-2 had been adapted to human cells from the start because it had been grown in humanized mice or in lab cultures of human cells, just as described in Daszak’s grant proposal (grants were assigned to Daszak of the EcoHealth Alliance, who subcontracted them to Shi; absurdly, Daszak was named by the WHO to investigate the virus origin). Wade further summarized that the invasion cannot begin until the S1 and S2 subunits of the spike protein have been cut apart. And there, right at the S1/S2 junction, is the furin cleavage site that ensures the spike protein will be cleaved in exactly the right place. The virus relies on the cell to do the cleaving for it. Human cells have a protein cutting tool on their surface known as furin. Furin will cut any protein chain that carries its signature target cutting site. This is the sequence of amino acid units PRRA at the core of furin cleavage site. And no known SARS-related coronavirus possesses a furin cleavage site. CoV-2 has the 12-nucleotide insert right at the S1/S2 junction. The insert is the sequence T-CCT-CGG-CGG-GC. The CCT codes for proline, the two CGG’s for two arginins, and the GC is the beginning of a GCA codon that codes for alanine. The double codon CGG-CGG for two arginine residues has not been found in any other beta-coronavirus (28). 

Other Scenarios

Nothing can be excluded at this stage. The above considered lab origin assumes genetic manipulations performed in the lab, but it might be possible that an isolated strain escaped from a lab without other lab manipulations. Additionally, the importance of recombination in the virus evolution is emphasized in many sources, as well as the mosaic character of the CoV-2 genome (22); recombination events might be involved even under lab conditions. The appearance of the omicron strain may also complicate the genetic relations among the known CoV-2 strains, as it seems rather different from the previous ones (41); the above-mentioned hypothesis of multiple origin comes to mind (40).

Without true information from the Chinese institutions, anything may be considered, including intentional release of the SARS virus or viruses. This act might be organized by any component of the highest leadership in China, comprising the Central Committee of the Communist Party or secret services. A perfidious eventuality of independently releasing two portions of somewhat different strains, possibly to confuse investigators may be theoretically also considered. The best way for China to gain the world’s trust will include providing true information, even if inconvenient for their self-image.

China's Deception and the Smoking Gun for the Lab Origin of COVID-19 

The U.S. government announced in their Fact Sheet of January 2021 that it had a reason to believe that several researchers inside the WIV had become sick in autumn 2019, before the first identified case of the outbreak (31). The Fact Sheet further noted that accidental infections in labs had caused several previous virus outbreaks in China and elsewhere, including a 2004 SARS outbreak in Beijing that had infected nine people, killing one. Starting in at least 2016 – and with no indication of a stop prior to the COVID-19 outbreak – WIV researchers had conducted experiments involving bat coronavirus RaTG13, identified by the WIV in January 2020 as its closest sample to SARS-CoV-2. The WIV had become a focal point for international coronavirus research after the 2003 SARS outbreak and had since studied animals including mice, bats, and pangolins. The WIV had a record of conducting “gain-of-function” research to engineer chimeric viruses. But the WIV had not been transparent or consistent about studying COVID-19 like viruses, or RaTG13 virus even after several miners died of SARS-like illness. For many years the United States had publicly raised concerns about China’s past biological weapons work, which Beijing had not documented, despite its clear obligations under the Biological Weapons Convention. Despite the WIV presenting itself as a civilian institution, it has collaborated with China’s military (31).

It can be added that China even disinforms about its real numbers of COVID-19 positive cases and deaths. So far they reported 74 cases and 3 deaths per million people, when the developed countries report between 20,000 and 300,000 cases per million and between 140 and 3500 deaths per million people in February 2022 (33). The total number of reported deaths in China has been constantly 4636 for a year now, which is neither compatible with other countries, nor with the reports about millions of fewer cellphone users in China already during the first three months of 2020 (16).

The odd insertion of the consensus recognition motif of the furin protease, PRRA, is recent since it is absent from all the close relatives of SARS-CoV-2. This observation is crucial as this site probably played a key role in the species barrier crossing and/or in the efficiency of human-to-human transmission, which is a prerequisite for the emergence of epidemics. The nucleotide codons coding for RR, namely the double codon CGG-CGG for two arginine residues, made the Nobel prize winner David Baltimore, eminent virologist and former president of CalTech, call it the smoking gun for the lab origin of the virus (28).  

After two years of the global paralysis, and as more than 400 million people around the world have been infected with SARS-CoV-2 and about 6 million have died, the world deserves the truth about the virus origin. An escape from one of the Wuhan laboratories seems to be the only feasible origin – until China does not provide reliable information supporting another scenario. 


References

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4 comments:

  1. As I am a non-expert, the information in the third paragraph from the end of the text is very important to me: "The total number of reported deaths in China has been constantly 4636 for a year now, which is neither compatible with other countries, nor with the reports about millions of fewer cellphone users in China already during the first three months of 2020."

    ReplyDelete
  2. You are right, one would hardly believe that, if one could not find the values at:
    https://www.worldometers.info/coronavirus/

    ReplyDelete
  3. Excellent post. I really enjoy reading and also appreciate your work.How Long Does Covid-19 Live On Fabric This concept is a good way to enhance knowledge. Keep sharing this kind of articles, Thank you.

    ReplyDelete
    Replies
    1. Thank you for your encouragement. I plan to summarize knowledge about additional aspects of the pandemic.

      Delete