Friday, August 14, 2020

Shi Zhengli disputes bats carry SARS-CoV-2, says whole genome sequence impossible, among other issues

Report by : Gan Yung Chyan, KUCINTA SETIA

Image : Video Screenshot


On 13 August 2020, MediaCorp Channel 8 News reported that Thai researchers have collected samples from local bats to see if the bat saliva contained SARS-CoV-2, hoping to help prevent and control the epidemic in Southeast Asia. 

The Thai researchers based on the above theory from "bat woman" Shi Zhengli's research team that claimed the expanded version of SARS virus originates from the Yunnan chrysanthemum horseshoe bat. However, with regards to this conclusion, Shi Zhengli has clarified in her latest interview with Science journal that SARS-CoV-2 does not evolve from RaTG13 the coronavirus which she identified from the bat saliva that her team collected in Yunnan and "patient zero" of COVID-19 (covid, in short) does not originate from Yunnan.

According to the Channel 8 report, the Thai researchers walked into a bat cave hidden deep in the dense forest of Sai Yok National Park in Thailand bordering Yunnan Province. They caught 200 bats with nets, took samples of bat saliva, blood and feces and released the bats later. They are testing whether these bats carry SARS-CoV-2.

Channel 8 reported that Shi Zhengli's team found the Yunnan horseshoe bat carried SARS-CoV-2 and Thai experts actually misunderstand that there is a high probability that the same species of bats in Thailand would also find the coronavirus.  

In response, Shi Zhengli disputed her earlier statements on SARS-CoV-2 in her latest interview with Science, according to Science & Technology Daily, a CCP Chinese scientific newspaper. The Wuhan Institute of Viology (WIV) virologist also confirmed that there are no wild bats on sale at Wuhan Huanan Seafood Market during the time of outbreak. She said bats do not carry SARS-CoV-2. Thus, Shi Zhengli hints that her statements have been misquoted by China CDC and China Communist Party (CCP) in early January until Zhong Nanshan announced in March that the place of origin of SARS-CoV-2 may not be in Wuhan. She also ruled out the possibilities of the bat that carried RaTG13, pangolins and cats as intermediate hosts. The mainstream media that keep hyping SARS-CoV-2 originating from bats and animals naturally serves to mislead people and propagate CCP's lie, shirking CCP's accountability for the pandemic.

Below are the consolidated translations of and explanations on Shi Zhengli's answers to Science's questions as reported by Science & Technology Daily.

Wuhan Institute of Viology's research work does not involve scale of viral transmission

Cross-species infections of viruses have always existed, and China has made arrangements for scientific research projects through the deployment of facilities, equipment and talent teams. With global climate change and the expansion and frequency of human activities, the risk of infection continues to increase. Wuhan Institute of Viology's research also confirms and supports this. Its research work does not involve the scale of viral transmission, Shi Zhengli claims.

There may be more than one intermediate host before the virus can infect humans

Shi Zhengli never said that SARS-CoV-2 definitely comes from bats. She said, "According to the research of our team and international colleagues, SARS-CoV-2 is likely to be derived from bats. After one or more intermediate hosts evolve, a virus that can adapt to humans and spread between humans will eventually appear. However, it is not clear which animal or which animal the intermediate host is and how it is transmitted to humans."

She adds, "There are two possibilities for cross-species transmission from natural host bats to humans. One is that bats directly transmit the virus to humans, and the other is that bat viruses are transmitted to one or more animal intermediate hosts and then to humans. For SARS-CoV-2, the first possibility cannot be ruled out, but it is very, very small. I prefer the second possibility."

The earliest cross-species transmission of SARS-CoV-2 from an intermediate host to humans, when and where it occurred, is currently not clear from scientific research. From historical experience, the initial place of outbreak of AIDS and other major diseases is not the origin (the initial interspecies transmission). This is also one of the important reasons why virus traceability is a challenging scientific problem. Regarding the source and transmission path of SARS-CoV-2, a broad perspective is required, and the joint efforts of scientists around the world are required, and it takes time.

Bats in Hubei do not carry coronavirus

WIV experts have been monitoring bat coronavirus in Hubei for many years. Shi Zhengli's team found that bats in Wuhan or even Hubei Province do not carry coronaviruses that are closely related to SARS-CoV-2. She thinks the possibility of the source of SARS-CoV-2 in Wuhan, Hubei, basically does not exist.

"Patient zero" does not come from Yunnan

Residents near the bat cave in Tongguan Town, Mojiang, Yunnan Province, are said to have visited Wuhan. So far, no nearby residents of the cave have been infected by SARS-CoV-2, and the so-called “patient zero” who lives near the mining area went to Wuhan is not true, Shi Zhengli declares.

SARS-CoV-2 in rolling door handles, floor and sewage of Wuhan Huanan Seafood Market

The early covid patients in Wuhan have nothing to do with the Huanan Seafood Market although Shi Zhengli is not aware that there are covid cases as early as November 2019 in Wuhan. She said,  "I did not participate in the epidemiological survey and I am not clear about this aspect. Our laboratory received clinical samples of "unexplained pneumonia" for the first time on 30 December."

WIV has detected SARS-CoV-2 nucleic acid in environmental samples such as the rolling door handles, floor and sewage of the Huanan Seafood Market. Shi Zhengli asserts, "We did not detect SARS-CoV-2 nucleic acid in frozen animal samples. This market may be the place where the early crowds of covid gathered and SARS-CoV-2 further transmission broke out."

Under the deployment of relevant departments of the Hubei Provincial Government, Huazhong Agricultural University and Shi Zhengli's team have collected environmental samples and frozen animal samples at the Huanan Seafood Market. SARS-CoV-2 nucleic acid were only detected in environmental samples such as rolling door handles, ground and sewage, and the number of genome copies is very low.

Not aware of animal-to-human viral transmission starting in Hubei or elsewhere

When asked if she is aware of the possibility of animal-to-human transmission starting in Wuhan or Hubei or in another city, Shi Zhengli said, "I can't judge until there is a definite answer. Virus tracing is a scientific problem, and scientists must speak with scientific facts and evidence. But the historical experience mentioned above is worthy of attention."

SARS-CoV-2 does not evolve from RaTG13 and there may be more than one intermediate host

The Science reporter asked Shi Zhengli, "You mentioned the bat coronavirus RaTG13 before. Obviously, the virus is the "ancestor" of SARS-CoV-2, and there is a 1100 nucleotide difference between the two. Other researchers used molecular clocks to make this judgment: RaTG13 and SARS-CoV-2 had a common ancestor at least 20 years ago. Do you have your own opinion on this? (For this epidemic), do you think there is an intermediate host between bats and humans? If it exists, which species do you think is most likely? Why? There are relevant data indicating that pangolin may be an intermediate host. What do you think of this?"

Shi Zhengli replied, "We have not received professional training in bioinformatics, nor have we calculated the evolutionary distance between RaTG13 and SARS-CoV-2. I think the evolution from the coronavirus carried by the natural host to SARS-CoV-2 should be spread through one or more intermediate hosts."

She added, "The genome sequences of the coronavirus, RaTG13 and SARS-CoV-2 carried by pangolins are relatively close and have a common ancestor. Judging from the current data analysis, I cannot determine whether the pangolin is a natural host or an intermediate host."

Cat may not be the intermediate host of SARS-CoV-2

On whether cats are likely to be the intermediate host, Shi Zhengli said that cats are not likely to be the intermediate host. She said, "The antibody test results of cat serum samples we collected after the outbreak of covid showed that the proportion of domestic cats infected with SARS-CoV-2 and their antibody levels were higher than that of stray cats. So I think that SARS-CoV-2 in cats is very likely to be spread by humans."

She further added, "Whether cats are the intermediate hosts of SARS-CoV-2 needs further research. Our team did not continue this research. From the current research conducted by us and other teams, cats may be infected by humans."

Is accidental virus infection in Wuhan Institute of Viology possible?

The Science reporter asked, "Many scientists have analyzed the sequence of SARS-CoV-2 and believe that the virus does not have the characteristics of artificial manufacture in the laboratory. However, some of these researchers said that there is still a possibility that the SARS-CoV-2 virus has appeared in your laboratory and accidentally infected a laboratory worker. They pointed out that it was precisely because of accidental virus infections that occurred in some laboratories that eventually led to the outbreak of SARS. How can you rule out this possibility?"

Shi Zhengli replied, "The virology research and experiment activities carried out by the Wuhan Institute of Viology are carried out in the corresponding biosafety laboratories in strict compliance with international and national biosafety laboratory management and experimental activity management requirements. Among them, P3 and P4 laboratories are very strict from facility to management. For example, scientific researchers must wear personal protective equipment, the air in the laboratory must be efficiently filtered before it can be discharged, and waste water and solid waste must be sterilized by high temperature and high pressure. The whole process of experimentation activities carried out by experimenters will be monitored by biosafety managers through video. The facilities and equipment of the laboratory and the implementation of the biosafety management system must be supervised and reviewed by the relevant professional departments of the country every year, and the operation can be continued after passing the review. The virus institute's high-level biosafety laboratory has been operating in a safe and stable state. So far, there has been no pathogen leakage or personal infection accident."

SARS-CoV-2 from RaTG13 is only theoretically possible

Shi Zhengli explains that the evolution of SARS-CoV-2 from RaTG13 is only a theory. It can be derived from another bat coronavirus. 

According to Shi Zhengli, among the three SARS-like live coronaviruses isolated in WIV's laboratory so far, the closest to SARS-CoV-2 has a genome similarity of only 79.8%, which is very significant. She said, "On 3 February, we published a paper in the journal Nature and reported a bat coronavirus (named RaTG13) that was 96.2% similar to SARS-CoV-2."

Coronavirus is one of the largest animal RNA viruses, with a genome of 30,000 nucleotides long. The 3.8% difference in genome sequence is a significant difference for coronaviruses. 

On 18 March 2020, five well-known scholars from the Scripps Research Institute, Columbia University, Tulane University, the University of Edinburgh in the UK, and the University of Sydney in Australia published a report entitled “SARS-CoV-2” in “Nature Medicine”. The article “Possible Path of Origin” pointed out that the difference between RaTG13 and SARS-CoV-2 in the receptor binding region is still very obvious. 

On 23 April 2020, the U.S. VOX News website quoted Edward Holmes, a virus evolution expert at the University of Sydney, who believed that the difference in the genome of SARS-CoV-2 and RaTG13 is equivalent to the difference in differentiation produced by the natural evolutionary process of about 50 years. 

Shi Zhengli said, "There are 1177 different sites in the genome of RaTG13 and the novel coronavirus carried by bats. It takes a long process to accumulate a sufficient number of mutations through natural evolution, and the probability that the mutations generated are highly consistent with the corresponding sites of the new coronavirus is very small. Therefore, The evolution of RaTG13 into SARS-CoV-2 in nature is only theoretically possible."

Does Wuhan Institute of Viology breed SARS-CoV-2?

The Science reporter asked, "Those who hold this view (accidental virus infection) also proposed several ways for the virus to spread from the Wuhan Institute of Virology. I would like to ask some practical questions about the working conditions in your laboratory, hoping to be more helpful to understand the above situation. Does your research institute breed bat coronaviruses?"

Shi Zhengli did not reveal about the working conditions in her laboratory. Instead, she answered, "We have only isolated three bat SARS-like live coronaviruses, which are 95%-96% similar to SARS virus genome, and are less than 80% similar to SARS-CoV-2. The results were published in Nature [2013, 503(7477):535-538], Journal of Virology [2016, 90(6), 3253-3256] and PLoS Pathogens [2017, 13(11): e1006698]."

Genome sequencing done in Wuhan

When asked if genome sequencing of SARS-CoV-2 is done in Wuhan or elsewhere, Shi Zhengli replied, "We isolate viruses or extract nucleic acids from samples, mainly to complete the sequencing work locally in Wuhan."

Results of animal experiments on SARS-related viruses to be released soon

The Science reporter asked Shi Zhengli another two questions, "Has your laboratory conducted animal experiments on SARS-related viruses recently? If it does happen, can you tell me the details?"

Shi Zhengli confirms that Wuhan Institute of Viology has done animal experiments on SARS-related coronavirus in 2018 and 2019. She claims the coronavirus used is "a bat SARS-like coronavirus that is similar to the SARS virus". She did not reveal if the bat coronavirus is the Zhoushan bat coronavirus or RaTG13. However, this statement may not suggest the possible presence of novel coronavirus SARS-CoV-2 in the laboratory since she has ruled out SARS-CoV-2 originates from bats and the laboratory only receives the sample which they named SARS-CoV-2 on 30 December 2019 from Wuhan Jinyintan Hospital.

Shi Zhengli said, "We have done genetically modified (human ACE2) mouse infection experiments and civet cat infection experiments in the Institute’s Biosafety Laboratory from 2018 to 2019. The virus used is a bat SARS-like coronavirus that is similar to the SARS virus. The relevant work is strictly in accordance with China's regulations on the biosafety management of pathogenic microorganism laboratories and other regulations are implemented. The results showed that bat SARS-like coronaviruses can directly infect civet cats, or they can infect mice through human ACE2 receptors but it is low in pathogenicity to mice and not to civet cats. The results are being compiled and will be announced to the public as soon as possible." 

Denies infection when collecting samples or processing bats recently

The reporter asked, "Is there such a possibility that the relevant personnel of the research institute were infected with the SARS-CoV-2 through other means, such as collecting samples or processing bats?"

Shi Zhengli replied, "What you said may never happen. We recently tested the serum of all laboratory personnel, and none of them were infected with bat SARS-like coronavirus and SARS-CoV-2. So far, all personnel in the institute have had zero infections."

Shi Zhengli suggested in the above that it is possible for the bat experts to be infected with SARS-like coronaviruses in the past but not lately.

Among 2007 bat samples positive for coronavirus, only RaTG13 closer to SARS-CoV-2

The reporter asked, "Do you have biological samples collected from bats in your laboratory, but have not been used for virus testing? If this is the case, how many samples have you tested and how many have not yet been tested? If some samples have not been tested, how can you be sure that they do not contain SARS-CoV-2 or other similar viruses?"

Shi Zhengli said, "We tested all the bat samples collected, including bat anal swabs, throat swabs and stool samples, of which 2007 were positive for coronavirus. Except for RaTG13, no coronaviruses closer to the SARS-CoV-2 gene sequence were found."

Genome sequence released to WHO on 12 January 2020

Shi Zhengli's team obtained the first batch of samples from 7 patients of Wuhan on 30 December 2019. Through the coronavirus universal RT-PCR method and the quantitative RT-PCR method that can detect all SARS-related coronaviruses, they found that 5 of the patients' samples were positive. On 31 December, when they sequenced the amplified products of the coronavirus by universal RT-PCR, they found that it was a new type of SARS-related coronavirus. In the meantime, after data analysis and multi-party verification, they performed whole genome sequencing and virus isolation, and on 12 January, they released the whole genome sequence of the virus to the world through WHO in time.

Discovery of RaTG13 in Mojiang Cave, Yunnan and its genome sequence

The reporter wanted to know about the discovery of RaTG13 in Mojiang Cave, Yunnan. He asked, "Please talk about your discovery in Mojiang Cave in 2013. When was the first time you did virus isolation? When did you get the full-length sequence of RaTG13?"

Shi Zhengli answered, "We obtained this sample from a bat dung swab in Tongguan Town, Mojiang County, Yunnan Province in 2013, and obtained a partial sequence of RdRp through the Coronavirus Universal Nucleic Acid Test (RT-PCR). Because this sequence is not very similar to SARS coronavirus, we did not pay special attention to this sequence."

RaTG13 or no RaTG13 sample in Wuhan Institute of Viology is a conflict

Shi Zhengli added, "In 2018, with the improvement of sequencing technology capabilities, we sequenced the remaining samples and obtained the whole genome sequence of RaTG13 except for the 15 nucleotides at the 5 ends. Since the samples were used to extract viral nucleic acid multiple times during the experiment, after the completion of the viral genome sequencing, the samples were no longer stored in the laboratory, and further studies such as virus isolation were not carried out."

Shi Zhengli claims, "In addition, among the many bat samples collected in the laboratory, only the RaTG13 virus was detected in one sample. In 2020, after obtaining the sequence of SARS-CoV-2, we compared it with all unpublished sequences and found that RaTG13 has a 96.2% sequence identity with SARS-CoV-2. This virus has never been isolated and cultured."

The above claim differs from Wang Yanyi, the director of the Wuhan Institute of Viology's P4 laboratory. Yanyi said that the laboratory does not have the RaTG13 virus sample at all. There is the conflict here. Did the virus institute have RaTG13 or not? Yan Limeng, the virology expert that defected to the US, and some other experts declare that RaTG13 is artificial since the Wuhan laboratory does not have a sample. How to conclude that SARS-CoV-2 comes from nature?

Only sequence of one protein, about naming virus samples

The reporter inquired, "Some people suspect that an experimental accident occurred in your laboratory, and believe that the bat virus you called BtCoV/4991 in 2016 is SARS-CoV-2. The report you published indicated that you only have the sequence of one protein, that is, RNA-dependent RNA polymerase (RdRp). According to the BLAST (Basic Local Alignment Search Tool, a set of analysis tools for similarity comparison in protein databases or DNA databases) carried out by GenBank, the analysis showed that the RdRp of BtCoV/4991 and RaTG13 have 100% homology. According to the description in your 2020 report, you have performed full-length genome sequencing of a virus that previously only obtained the RdRp gene sequence-can you consider BtCoV/4991 to be RaTG13? If this is the case, why do you want to rename the virus? What does "TG" in RaTG13 stand for?"

Shi Zhengli replied, "Ra4991 is the number of a field sample, and RaTG13 is the number of the coronavirus detected in the sample. When we changed the name, we mainly considered the time and place of the sample. 13 is the year 2013 of the sample, and TG is the acronym for customs clearance."

Whole genome sequence of SARS-CoV-2 impossible

The reporter asked further, "Why do you only have the RdRp sequence of some viruses instead of the complete sequence? How many complete sequences are there in the samples you have tested, and which samples only contain RdRp sequences?"

Shi Zhengli admits that Wuhan Institute of Viology has constraints and it is not possible to obtain the whole genome sequence of SARS-CoV-2. She said, "Due to financial and manpower constraints, it is impossible for us to complete the determination of the whole genome sequence of all samples. We hope to complete the determination of the whole genome sequence of coronavirus in some samples within the next 2 years. It should be pointed out that the amount of nucleic acid in some samples is very low and it is impossible to obtain the whole genome sequence."

Shi Zhengli claims no virus destruction

The reporter asked, "After the SARS-CoV-2 outbreak, have you received instructions to destroy certain viruses?" Shi's answer is "No".

Is accidental spread from other laboratories possible?

The reporter asked, "Is it possible that the virus accidentally spread from other laboratories in Wuhan, for example, the Wuhan Center for Disease Control and Prevention? If you think there is no such possibility, what is the basis?" Shi Zhengli has ruled out the possibility. She said, "Based on the daily academic exchanges and discussions, I think there is no such possibility."

Coronavirus research done in P2 or P3 laboratory, SARS-CoV-2 experiments in P4

The reporter asked, "Given that most of the coronavirus research in places is done in BSL-2 or BSL-3 laboratories, in fact, Wuhan Institute of Virology did not have an operational BSL-4 laboratory until recently. Why would you be in BSL-4 conduct coronavirus experiments under laboratory conditions?"

Shi Zhengli denies that her coronavirus research is executed in the P4 laboratory. She said, " The coronavirus research in our laboratory is carried out in the BSL-2 or BSL-3 laboratory, not in the BSL-4 laboratory."

After the institute’s BSL-4 laboratory (P4 laboratory) was put into operation, in accordance with the management regulations of the BSL-4 laboratory, we used the low-pathogenic coronavirus as a model virus in the BSL-4 laboratory to train scientific researchers for the purpose of preparing for experimental activities of highly pathogenic microorganisms.

After the outbreak, the state clearly stipulated that the novel coronavirus cultivation and animal infection experiments should be carried out in biosafety level three and above laboratories. Because the BSL-3 laboratory of the institute does not have the hardware conditions for conducting experiments on non-human primates, in order to carry out related research, the institute applied to the relevant state departments and obtained the qualification of the BSL-4 laboratory to carry out SARS-CoV-2 experiment activities, and carried out rhesus monkey animal model experiments.

Relevant experimental activities are supervised by the Institute's Biosafety Committee and comply with biosafety management regulations.

Shi Zhengli's cooperative research

With the emergence of various emerging and sudden infectious diseases around the world, scientists from various countries have a comprehensive understanding of the situation of wild animals carrying viruses. This is not only the key to early warning of emerging and sudden infectious diseases from the source, but also an important part of the prevention and control of infectious diseases on a scientific basis. 

Shi Zhengli said that she has conducted cooperative research with Peter Daszak, Chairman and Chief Scientist of the American Ecological Health Alliance, and established a good cooperative relationship in the fields of virus monitoring and new pathogen discovery. The research team found a variety of coronavirus with different sequences in the bat population. Some of these coronaviruses have potential cross-species transmission to humans and animals, such as SARS, MERS, SADS etc., which are related infections. Disease prevention and control provides important clues.

NIH suspends funds to Wuhan Insititute of Viology

Regarding the US National Institute of Health (NIH)'s suspension of funding for cooperation projects between the two parties, the virus institute staff do not understand and feel very sorry. This project is supposed to be an international cooperation project that will help to unite scientists around the world to jointly carry out early warning and prediction of infectious diseases, and will further promote the development of related vaccines and drugs to protect humanity from the threat of coronavirus.

Monitoring coronavirus for 20 years, further cooperative research needed

Shi Zhengli has been working in the field of viology for 20 years monitoring the impact of the coronavirus on human health. She said, "Based on the experience of the past 20 years, the coronavirus has been interfering with and affecting the normal production and life of human beings."

She called on the world to further strengthening international cooperation in the traceability of emerging infectious diseases and other viruses. She hopes that scientists from various countries will work together to carry out related research. She concluded, "The purpose of virus traceability is to prevent the recurrence of similar epidemics from harm to human society, and humans can respond more effectively after the epidemic arrives."

Source : Science & Technology Daily via ScienceNet

Ref: http://news.sciencenet.cn/htmlnews/2020/8/443910.shtm

Other Refs:

Yan Limeng: HCQ, vaccines cannot work if 'lab-modified" nature of SARS-CoV-2 is disregarded, https://staygate.blogspot.com/2020/08/yan-limeng-hcq-vaccines-cannot-work-if.html

Wuhan Institiute of Viology does not have RaTG13 in the viral genome of SARS-CoV-2 which is "not natural", https://staygate.blogspot.com/2020/05/wuhan-institute-of-viology-does-not.html

Shi Zhengli worried about laboratory virus leakage, https://staygate.blogspot.com/2020/04/shi-zhengli-worried-about-laboratory.html

Shi Zhengli : Unexpect SARS-like coronavirus cross-specifies infection will occur in Wuhan, https://staygate.blogspot.com/2020/03/shi-zhengli-unexpect-that-sars-like.html

External Ref:

Bernardo Cervellera, Prof Tritto: COVID-19 was created in the Wuhan laboratory and is now in the hands of the Chinese militaryhttp://www.asianews.it/news-en/Prof-Tritto:-COVID-19-was-created-in-the-Wuhan-laboratory-and-is-now-in-the-hands-of-the-Chinese-military-50719.html?fbclid=IwAR3XFwlekdGVKXjUYkKNi094S_qnrXnjb1SSJNzTBG7D3oMN_Zxj3cRM6q8


https://www.youtube.com/watch?v=z95KJCxDHX0


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