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Billy Bostickson 🏴👁&👁 🆓 Profile picture
@BillyBostickson
Nov 16, 2021 • 56 tweets • 38 min read • Read on X
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1. "The Bats Down Under" #CSIRO Thread

CSIRO = Commonwealth Scientific and Industrial Research Organisation,

Australian Government . Image
2. Location of the Labs

google.com/maps/d/u/0/vie… Image
3. Australian Acronyms

Australian Centre for Disease Preparedness (ACDP)

The Australian Animal Health Laboratory (AAHL)

Page 21

apps.who.int/iris/bitstream… Image
4. Summary from Attachment 1.2

Australian Animal Health Laboratory (AAHL)

bwc-ecbm.unog.ch/system/files/f… ImageImage
5. Facilities and Collections
csiro.au/en/about/facil…
6. How it all started #CSIRO



"..using the world renowned bat virus isolation methodology developed by scientists at CSIRO’s Australian Centre for Disease Preparedness in Geelong"

web.archive.org/web/2014082503…

csiro.au/portals/media/… ImageImage
7. The Paper & Co-Authors (CSIRO, EHA, WIV)

Bats Are Natural Reservoirs of SARS-Like Coronaviruses (2005)

science.org/doi/10.1126/sc… Image
8. Turn of the Century - #CSIRO Dreams

10 minute video on @CSIRO and China

scienceimage.csiro.au/video/11899/wo… Image
9. Old school computers and real telephones

"Ni Hao Peng You, How are things?" ImageImage
10. 2006

11. 2010

12. Peng Zhou 2011

13. Nipah Thread 2011 - 2020 #CSIRO #WIV #EHA

14. 2012 #HENDRA

Bat virus could hold key to Hendra

“Field work with bats is an essential part of research into identifying new viruses,” Dr Hume Field of Biosecurity Queensland said

blog.csiro.au/bat-virus-coul…
15. 2012. Long Life? Wishful Thinking?

The Bat Pack with Beijing Genome Institute, sequenced the genomes of 2 bat species:

Black Flying Fox, Australian megabat
David’s Myotis, Chinese microbat

blog.csiro.au/long-life-and-…

csiropedia.csiro.au/evolution-of-f… ImageImageImageImage
16. 2013 Paper

Comparative Analysis of Bat Genomes Provides Insight into the Evolution of Flight and Immunity

pubmed.ncbi.nlm.nih.gov/23258410/

Available thanks to @SciHub_

sci-hub.ee/10.1126/scienc… ImageImage
17. Interlude "The Man in the Suit"

Shawn Todd (A.K.A. Strawnie) hard at work in the highly secure PC4 lab

blog.csiro.au/the-man-in-the…
18. 2013 Article by Linfa Wang & Gary Crameri

what are zoonotic diseases?

blog.csiro.au/first-hendra-n…

Original Article:

theconversation.com/first-hendra-n…

Bio of Linfa Wang

2005–2011
Honorary Professor, Wuhan Institute of Virology

2008–2011 CSIRO

theconversation.com/profiles/linfa…

(Pic=Marburg) ImageImageImage
19. 2013. What's Next?

There is a growing view that a One Health approach will be critically important for our preparedness for the next zoonotic pandemic.....NOT!

blog.csiro.au/human-spillove… Image
20. 2013 How to stop an epidemic

by Linfa Wang #CSIRO

blog.csiro.au/how-to-stop-an…
21. Hendra Vaccine developed by #CSIRO

"For instance, in 2012 we launched the Equivac vaccine, which protects horses from infection and thus prevents transmission of Hendra virus from horses to humans" ImageImageImage
22. Hendra Vaccine - Lawsuits

1. lhd.com.au/class-actions/…
2. theland.com.au/story/6355621/…
3. horsetalk.co.nz/2018/03/22/cla…
4. horsesandpeople.com.au/hendra-vaccine…

"The 1st test of the vaccine started in 2012 & animal vaccine manufacturer Zoetis (formerly Pfizer) obtained the rights to manufacture it" Image
23. 2014 - Ebola

From animals to humans: understanding Ebola virus

"Fortunately Australia has a robust system to deal with an emergency disease outbreak, including our very own Australian Animal Health Laboratory"

blog.csiro.au/from-animals-t… Image
24. 2014 #WIV & #CSIRO & IRF7

Peng Zhou, Linfa Wang, Zhengli Shi, Lijun Wu (WIV)



IRF7 in the Australian Black Flying Fox, Pteropus alecto: Evidence for a Unique Expression Pattern & Functional Conservation

journals.plos.org/plosone/articl… Image
25. 2014 paper based on work dating back to 2011



Type III IFN Receptor Expression and Functional Characterisation in the Pteropid Bat, Pteropus alecto

proquest.com/docview/130880… Image
26. 2014 - Bat’s immunity may hold key to preventing future Ebola outbreaks (Michele Baker)

If we redirect the immune responses of other species to behave the same way as bats, the high death rate of diseases such as Ebola could be a thing of the past

blog.csiro.au/bats-immunity-…
6 years left to finish, break time, Image
27. Rewind to 2012 #CSIRO

Antiviral Immune Responses of Bats: A Review

M. L. Baker, T. Schountz and L.‐F. Wang

ncbi.nlm.nih.gov/labs/pmc/artic…

Mentioned here:

ImageImage
28. Fast Forward to 2016

24/7 immunity from lethal diseases? It’s in the bat, man!

blog.csiro.au/bat-disease-im… Image
29. Flying Foxes #CSIRO

"In a paper in PNAS, Michelle & her team examined this evolutionary cheat code in Australian black flying foxes"

refers to Zhou et al (2016)

Contraction of type I IFN locus & unusual constitutive expression of IFN-α in bats

pnas.org/content/113/10… Image
30. The Question of Live Bats at #WIV



Scroll up thread for more details of live bats at WIV

1. Patents
2. Video
3. Anecdotal Evidence
31. Peng Zhou and Zhengli Shi #STING



Why Bats Make Such Good Viral Hosts

The bat version of the STING protein helps dampen the mammals' immune response to infection, researchers have found.

archive.ph/7ygOO
32. #WIV Thanks for your help #CSIRO Zai Jien!

Dampened STING-Dependent Interferon Activation in Bats

cell.com/cell-host-micr…

What the Aussies taught them came in handy later on

ImageImage
33. Summary so far (1)

1. CSIRO helps "developing' China with Biosafety
2. CSIRO teaches Chinese bat virus isolation methodology (Tweet 6)
3. CSIRO - WIV - EHa SARS Paper (2005) Tweet 7
4. PZ, ZLS & LFW at CSIRO
5. PZ & LFW Human & Bat Toll Like Receptors - 2010 (Tweet 11)
34. Summary so far (2)

6. Nipah, Ebola Research and Hendra Vaccine (CSIRO)
7. Bat Immune system & longevity research at CSIRO
8. Bat immune system & longevity research at WIV
9, Live Bat Colony at WIV (and Singapore under LFW)
35. Live Bats at WIV - Myotis Davidii

Why did WIV keep Myotis species bats at WIV in 2017?

Because of their longevity (lifespan up to 37 years)

Growing old, yet staying young: The role of telomeres in bats’ exceptional longevity

readcube.com/articles/10.11… ImageImageImage
36. WIV worked with wild-caught out bred bats and their own colony of live bats (Myotis) as well as splenocytes, tissue and cell lines from these bats.

Dampened STING-Dependent Interferon Activation in Bats
cell.com/cell-host-micr… (2018) Image
37. The Problem

first Identified here:



(Banarjee et al, 2018) + Michelle Baker, CSIRO

Tools to study pathogen-host interactions in bats

ncbi.nlm.nih.gov/labs/pmc/artic… Image
38. Repeat (1)

Primary cells can also harbor latent viruses that can become reactivated during in vitro cultivation when the cells are outside the host and isolated from other components of the immune system that would otherwise control virus replication
39. Repeat (2)

"Cell lines from any species of bat should therefore only be handled under controlled biosafety conditions and thoroughly tested before being used for experimental studies. This is also a limitation when working with wild-caught bats"
40. Graphic from Paper

Tools to study pathogen-host interactions in bats

ncbi.nlm.nih.gov/labs/pmc/artic… Image
41. Live Bats at #WIV

In addition to cell lines, there is a need for lab bred bats with known infection status for ex & in vivo experiments. Establishing a wild-caught bat colony is challenging but achievable with sufficient resources to house the bats

Image
42. 2015 #WIV Bat Cell Line Experiments

Cell lines used:

1. M. davidii primary kidney cell line (MdKi)

2. immortalized Rhinolophus sinicus kidney cell line (RsKT)

Cloning, expression, antiviral activity of interferon β from Myotis davidii

ncbi.nlm.nih.gov/labs/pmc/artic… ImageImageImage
43. What experiments on these bat cell lines?

They were "attacked" with:

1, Bat adenovirus TJM (BtAdV-TJM)

2, Sendai virus (SeV)

3, Vesicular stomatitis virus

Image
44. Interlude - French Kissing Vampire Bats

Please do not try this at home

earthlymission.com/vampire-bats-f… Image
45. 2016: In Vivo versus In Vitro (PZ, LFW)



"When comparing in vitro studies in bat cell lines, bat viruses exhibited subversion of the bat immune system"

establishment of Pteropus alecto bone marrow-derived dendritic cells

ncbi.nlm.nih.gov/labs/pmc/artic… ImageImage
46. 2013. Peng Zou, LFW & Michelle Baker #CSIRO

Bat Mx1 and Oas1, but not Pkr are highly induced by bat interferon and viral infection

pubmed.ncbi.nlm.nih.gov/23541614/

(IFN-stimulated genes (ISGs) in Bats)

Freed PDF version thanks to @SciHub_

sci-hub.ee/10.1016/j.dci.…
47. 2015. Cloning, expression, & antiviral activity of interferon β from Chinese microbats

ncbi.nlm.nih.gov/pmc/articles/P…

Recombinant IFNβ inhibited vesicular stomatitis virus & bat adenovirus replication in cell lines from M. davidii & Rhinolophus sinicus.

Image
48. 2016/2017 CRISPR knockout bat cell lines

WIV and Duke NUS

impact of IFNAR2 knockout on virus infection (CRISPR/Cas9 bat cell lines)

ncbi.nlm.nih.gov/labs/pmc/artic…

Since then, we have made several other knockout bat cell lines (to be published elsewhere).

ImageImageImageImage
49. 2020. Accelerated viral dynamics in bat cell lines
...with implications for zoonotic emergence

Such rapidly-reproducing viruses would likely generate extreme virulence upon spillover to hosts lacking similar immune capacities to bats.

ncbi.nlm.nih.gov/labs/pmc/artic…

+ 3 videos ImageImageImageImage
50. From CSIRO to WIV and back to CSIRO

This thread began with CSIRO and revealed how CSIRO shared their techniques and research with Chinese Bat Researchers who went on to develop these techniques in China.

Now back to CSIRO - AAHL Lab

51. Beating the world's deadliest viral villains #CSIRO

BSL4 at AAHL

52. CSIRO AAHL Links
1, Geelong: Australian Animal Health Laboratory (AAHL)
web.archive.org/web/2007111418…
2. CSIRO's biotechnology research
web.archive.org/web/2007100518…
3. A look inside CSIRO's Australian Animal Health Laboratory
web.archive.org/web/2007111903…
Transcript:
web.archive.org/web/2017100716…
53, AAHL is built on a box-within-a-box principle

4. AAHL Video
facebook.com/watch/?v=31039…
5. AAHL brochure
web.archive.org/web/2007102710…
6. Under the hood of our biocontainment facility
blog.csiro.au/under-the-hood… ImageImageImage
54. Wise Advice from Michelle Baker

“An effective approach to maintaining our distance from bats is to avoid all physical contact with them by not handling live or dead bats.

blog.csiro.au/virus-mutation/

Unfortunately Bat Researchers in China failed to heed her advice... Image
unroll CSIRO - WIV Bat Cell Lines @threadreaderapp Image

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More from @BillyBostickson

Billy Bostickson 🏴👁&👁 🆓 Profile picture
Aug 13
Baric's hACE2 transgenic mice were shared with the WIV for GOF like virus experiments designed to "assess" the potential for coronaviruses to jump from animals to humans.

We asked how many were sent to WIV

researchgate.net/publication/35…

Record via @jhas5 here

arlo.riseforanimals.org/entity/univers… x.com/jhas5/status/1…
2. Baric spills his humanized mice beans to Father Racaniello during a heart rending TWIV confessional in March 2020, admitting he sent his hACE2 mice to WIV in 2018 (as part of the infamous NIAID grant mentioned above):



Timestamp; 01:00:30 - 01:03:15microbe.tv/twiv/twiv-591/
3. Daszak

"Our R01 work shows that bats in South China harbor many SARSr-CoVs (50+), some of which are able to use human ACE2 to enter human cells, infecting humanized mouse models to cause SARS-like illness, and evade available therapies or vaccines"

ecohealthalliance.org/wp-content/upl…
Read 9 tweets
Billy Bostickson 🏴👁&👁 🆓 Profile picture
Aug 11
1. New England Biolabs & Novagen

The amplified Rp3-S1 fragment was digested with BamHI & XhoI & ligated into the expression vector pMAL-C2X (New England Biolabs).

The 5 truncated fragments were digested with BamHI & XhoI and ligated into the expression vector pET32a (Novagen).
2. Promega & Takara

Protein expression & purification

After purification, recombinant plasmid was transformed into BL21 competent cells (purchased from Promega)

Recovered PCR product & expression vector pET32a were digested with restriction endonucleases BamHI & XhoI (Takara).
3. Takara (2)

After DNA recovery, the PCR product was ligated to the expression vector pET32a using T4 DNA ligase (Takara)
Read 32 tweets
Billy Bostickson 🏴👁&👁 🆓 Profile picture
Jul 26
@breakfast_dogs @gadboit @NLink247 1. Insect cell baculovirus cell cultures become contaminated with insect genomic material.
2. Replication process can lead to presence of viral DNA & cellular components in culture medium.
3. This LEADS to contamination of viral product with baculovirus & insect cell-derived DNA.
@breakfast_dogs @gadboit @NLink247 Contamination

4. Expression of SC-2 Spike RBD Using the Insect Cell-Baculovirus System

pmc.ncbi.nlm.nih.gov/articles/PMC97…

5. Co-expression vs. co-infection using baculovirus expression vectors in insect cell culture

pmc.ncbi.nlm.nih.gov/articles/PMC71…

6. insect cell lines
sciencedirect.com/science/articl…
@breakfast_dogs @gadboit @NLink247 Did WIV use insect cell systems with SARS-C0V-2?

7. Insect cell baculovirus expression systems

x.com/BillyBostickso…

8. DENG Fei, Chief Investigator in Research Group on Viruses Resources at WIV engineered baculovirus-insect cell expression systems.

buzzchronicles.com/b/all/17652/
Read 13 tweets
Billy Bostickson 🏴👁&👁 🆓 Profile picture
Jul 22
1. Kadlec awaiting confirmation for his new role

2. Kadlec's report references pangolins 76 times

bush.tamu.edu/wp-content/upl…

3. Time to read my pangolin coronavirus report?

researchgate.net/publication/39…

4. DRASTIC research cited
@gdemaneuf @ydeigin @BillyBostickson @C_Small_ x.com/BoDeepest/stat…
1. the article linked is quite good, more comments and analysis will be posted here soon

city-journal.org/article/robert…Image
2. Possible lab manipulations

"2) insertion of a pangolin virus recombinant spike or the spike’s RBD protein that includes a sequence for an FCS and an integrin-binding protein" Image
Read 76 tweets
Billy Bostickson 🏴👁&👁 🆓 Profile picture
Jul 9
Pressure is needed on people like @realannapaulina @RepLuna & @GOPoversight team as their mandate may dissolve in August.

They did Kennedy, but they are supposed to do UFO files & CoVid-19 origins before they dissolve.

We also need to know about the subpoenas sent by @RandPaul
What was promised and demanded (for the record)

oversight.house.gov/release/rep-lu…

oversight.house.gov/release/chairm…
If you are a concerned US citizen please directly contact the people listed here:

oversight.house.gov/subcommittee/t…
Read 8 tweets
Billy Bostickson 🏴👁&👁 🆓 Profile picture
Jul 1
🧵Rev. Billy's Lab Leak Cover-Up Sermon

1. Brethren, hear me now, for we stand at the precipice of a great & grievous iniquity!

The world has been ensnared by a monstrous conspiracy, a vile machination spun by a clique of scientists, experts, media scribes & state overlords!
🧵2. Let us never forget!

Those who, in their arrogance, have sought to veil the truth of the COVID pestilence!

And I say to you today, that this is no mere error, but a deliberate, diabolical plot to obscure the origins of a plague that has scourged the nations!
🧵3. The Vocabulary of Deception

Let us expose this falsehood, this house of deceit

& the duties of the vigilant in these perilous times!

Oh, what a wretched lexicon these deceivers employ! Their words are cloaked in the garb of science, yet they reek of falsehood & treachery!
Read 23 tweets

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