Dear Dr. Francis M. Jiggins, Since You Asked Openly
How Does Wolbachia Do What It Does?, I Have Some Thoughts
Credit: RoseWrites Own Zazzle Store Image | Fair Use Image of Dr. Francis M. Jiggins Bio Page, Cambridge University
Thank You For Asking
And For Keeping an Open Mind
On April 15th, 2018, I found your 2016 paper called Open questions: how does Wolbachia do what it does? and I may have some answers (or partial answers) for you.
Six Key Points in Your Paper That Deserve Answers
1) The bacterium could not be cultured.
Not true. The 2003 paper, Culture and Phenotypic Characterization of a Wolbachia pipientis Isolate, by Florence Fenollar, Bernard La Scola, Hisashi Inokuma, J. Stephen Dumler, Mark J. Taylor, and Didier Raoult states:
"Bacterial growth could be obtained in C6/36, another A. albopictus cell line, at 28°C and in a human embryonic lung fibroblast monolayer at 28 and 37°C [98.6 degrees F], confirming that its host cell range is broader than was initially thought."
2) Wolbachia-infected mosquitoes were being released to prevent the transmission of dengue virus. However, the mechanisms underlying these effects remain poorly understood.
I believe Wolbachia does not prevent the dengue virus (nor ZIKV and perhaps other viruses) from being acquired by the Aedes or the Culex spp. of mosquito. But rather, these smaller viruses "hide" within the Wolbachia (acting as bacteriophages).
Study Literature Which Supports My Theory
A 1999 paper, Replication of dengue type 2 virus in Culex quinquefasciatus (Diptera: Culicidae), by Vazeille-Falcoz M, Rosen L, Mousson L, and Rodhain F. states:
"The replication of dengue virus in Cx. quinquefasciatus was either at a very low level of magnitude or generated a large number of noninfectious particles."
"Several investigators reported the isolation of dengue virus from Cx. quinquefasciatus collected in nature, namely, in China, Vietnam, and New Caledonia. Furthermore, experimental studies carried out in China suggest that this species not only could become infected, but was also capable of the transmission of dengue viruses to mice."
"Graham, who first demonstrated the transmission of dengue by mosquitoes in 1902 – 1903, thought that Culex quinquefasciatus (formerly Culex fatigans) was the vector."
Why I Believe Graham Should Have Never Been Dismissed
When I read through scientific literature, I found there was a decided path taken to completely ignore Culex quinquefasciatus as a vector of dengue (and more recently, the Zika virus).
And to this day, the downplaying of Culex spp. as vectors of disease continues.
What also continues to spread, though, is the dengue virus and Zika virus — even in countries with world-class vector control, like Singapore.
The Question Was Asked Six Years Ago on ResearchGate
How Many Species of Mosquito Can Transmit Dengue Host-to-Host:
Credit: Fair Use Image of Research Gate Question and Answers
Credit: RoseWrites on InfoBarrel
What About "Large Number of Noninfectious Particles"?
What WAS Considered Noninfectious Turned out to Be Highly Infectious
The 2014 paper, Immature Dengue Virus Is Infectious in Human Immature Dendritic Cells via Interaction with the Receptor Molecule DC-SIGN, by Richter MKS, da Silva Voorham JM, Torres Pedraza S, Hoornweg TE, van de Pol DPI, Rodenhuis-Zybert IA, et al. confirms:
"Fully immature DENV particles are considered non-infectious, but antibodies have been shown to rescue their infectious properties. This suggests that immature DENV particles only contribute to the viral load observed in patients with a heterologous DENV re-infection."
"Furthermore, immature particles may be important in the initiation of infection as virus particles produced in mosquito cells are known to have a high prM content."
The Paper, Immature Dengue Virus: A Veiled Pathogen?
By Izabela A. Rodenhuis-Zybert, Hilde M. van der Schaar, Júlia M. da Silva Voorham, Heidi van der Ende-Metselaar, Huan-Yao Lei, Jan Wilschut, and Jolanda M. Smit
This 2010 paper confirmed, yet again, the downplaying of important findings:
"Interestingly, however, numerous functional studies have revealed that immature particles lack the ability to infect cells."
These researchers concluded: "Together, these data suggest that during a secondary infection or primary infection of infants born to dengue-immune mothers, immature particles have the potential to be highly infectious and hence may contribute to the development of severe disease."
The 2015 paper, Modeling the indirect effect of Wolbachia on the infection dynamics of horizontally transmitted viruses, by Jakob F. Strauß and Arndt Telschow opened my eyes to how linear thinking would lead many scientists down the wrong path. Note this line:
"It was shown that the presence of a virus facilitates the invasion of Wolbachia ..."
Remember What Happens in the Natural Environment
Some Mosquitoes (+/- Virus) Will Naturally Acquire Wolbachia
Credit: Bouncer by Incase on flickr (CC-by-2.0) | Happy Bouncer by Susan Sermoneta on flickr (CC-by-2.0) | Text by RoseWrites
But I've Been Told Only Aedes Transmits Dengue and Zika
The fact that Wolbachia enhances pathogens in the Culex spp. caused me to draw the immediate conclusion that these smaller viruses must be hiding and replicating within Wolbachia. Ergo, Culex spp. can transmit both the Wolbachia and the virus.
And why wouldn't it? If Culex spp. transmits the nematode that emits Wolbachia, it makes sense that Wolbachia could pass through the proboscis of the Culex spp. (even if it cannot in the smaller Aedes species of mosquitoes).
And the bacterium, Wolbachia, would provide most (if not all) of what the dengue and Zika virus requires for replication. It would certainly explain the following:
1) The presence of a [what was touted as] a large number of noninfectious particles.
"In one patient, we found compartmentalized strains: the virus present in his semen was different from the virus in his urine. In all cases, the pathogen we found in the final stage of the infection wasn’t the same as the virus that entered the patient."
3) ZIKV found in pools of whole male mosquitoes (Aedes aegypti and Culex quinquefasciatus).
The largely ignored 2018 paper, Zika Virus in Salivary Glands of Five Different Species of Wild-Caught Mosquitoes from Mexico, by D. Elizondo-Quiroga et al. clearly states:
"... the fact that pools of male mosquitoes were found to be positive for ZIKV, suggests vertical transmission in these species."
Simply put: If male mosquitoes do not blood feed, then how would they acquire a virus easily without the assistance of a bacterium that is ALSO maternally inherited?
Oxitec's Vector Control Solution 28-Page PDF Highlights
The Dangers of MosquitoMate's Wolbachia-Infected Aedes Releases:
Credit: Fair Use Portion of PDF by Oxitec, Ltd.
Downplay Culex as a Vector; Downplay Immature Virions
Add Some Folks Who Ask In A Forum If Culex Matter = Groupthink
On the surface, it might appear that Wolbachia is inhibiting these viruses when in fact, it's hiding them.
When I looked for any evidence of this, I did indeed find it. See screenshot above of PDF snippet.
Since we have not been able to get control over dengue, despite many measures, I believe the Wolbachia being used in Aedes is making disease transmission worse, not better. Particularly when the Culex spp. naturally acquires Wolbachia.
And mosquitoes can carry more than one strain of Wolbachia, which makes this even more frightening.
Similarly, the 2012 paper, West Nile Virus: Biology, Transmission, and Human Infection, by Tonya M. Colpitts, Michael J. Conway, Ruth R. Montgomery and Erol Fikrig stresses:
"Immature or partially mature flavivirus particles of both DENV and WNV have been shown to account for up to as much as 40% of the total virus population in a given infection. While they were traditionally thought to be noninfectious, several recent studies have shown that immature WNV particles can be highly immunogenic and infectious in vitro and in vivo when bound by antibodies against the E or prM protein."
3) The bacterium modifies the sperm of infected males during spermatogenesis so that the paternal chromosomes condense when an egg is fertilised, which typically kills the developing embryo.
I believe vertebrate species are suffering mass breeding failures because of Wolbachia-infected Aedes releases (carried out by the trillions since 2009, knowingly). It would explain the following:
Credit: Rose Webster (aka RoseWrites on InfoBarrel)
4) Despite detailed descriptions of how CI disrupts the cell cycle and paternal chromosomes, the molecular basis for how paternal chromosomes are marked and then rescued remains unknown. Advances in epigenetics and chromosome biology make this a timely moment to return to this question.
"... that a change of a single amino acid — likely occurring sometime in 2013 — created a new strain of Zika much more dangerous to developing brain cells."
And it was then that everything clicked. Zika became ugly in French Polynesia, causing a 20-fold increase in Guillain-Barré syndrome (GBS).
I thought: Could it be that the Wolbachia might become airborne and infect human lung cells? And could Zika be the phage that enables Wolbachia to infect mammals with ease?
Keep in mind, a 2012 paper, Postpartum Group A Streptococcus Sepsis and Maternal Immunology, by Katie L. Mason and David M. Aronoff emphasized:
"Notably, there does not appear to be an increase in GAS [Group A Strep] antibiotic resistance, so other factors must underlie the re-emergence of GAS postpartum infections."
And a brilliant 2005 paper, The microbial pan-genome, by DuccioMedini, ClaudioDonati, HervéTettelin, VegaMasignani, and RinoRappuoli made me realize now how vulnerable we will be because Wolbachia is in our environment in massive quantities:
"... bacterial species will never be fully described, because new genes will be added to the genome of the species with each new genomic sequence."
"Although new genes can originate through duplication of existing sequences, followed by diversification, the most common way to acquire new functions is by the transfer of genetic material from unrelated organisms."
UNRELATED organisms could mean Wolbachia (Rickettsia) and Zika alongside strep A or simply the normal bacteria that we all carry.
Credit: Fair Use Portion of The Microbial Pan-Genome by DuccioMedini, ClaudioDonati, HervÃ©Tettelin, VegaMasignani, and RinoRappuoli
Credit: Fair Use Portion of The Microbial Pan-Genome by DuccioMedini, ClaudioDonati, HervÃ©Tettelin, VegaMasignani, and RinoRappuoli
5) However, it remains unclear whether different processes may account for the breadth of Wolbachia’s antiviral effects, which operate against a broad range of viruses in a diverse range of insects. The exact mechanism by which Wolbachia prevents viral replication remains unknown.
When Wolbachia is hiding the virus, it might appear (in Aedes) that the virus was inhibited, right?
In Culex, however, both Wolbachia and the viruses (dengue and Zika) would be transmitted. If the worm that emits Wolbachia can travel through the proboscis of Culex, I assume Wolbachia can too.
The 2013 study, High-Efficiency Thermal Asymmetric Interlaced PCR (hiTAIL-PCR) for Determination of a Highly Degenerated Prophage WO Genome in a Wolbachia Strain Infecting a Fig Wasp Species, made me think again of ZIKV (dengue, and other viruses) as a phage too. It states:
"... one phage could potentially be used to modify a broad range of Wolbachia stains (Tanaka et al., 2009; Kent and Bordenstein, 2010;Wang et al., 2013)."
ADDENDUM: May 1st, 2018
What confirmed my worst fear was a study sent to me by Anon E. Moose in the comments section of my paper Why We Need to Investigate Wolbachia-Infected Mosquito Releases:
Published April 16th, 2018, the study Variation in Wolbachia effects on Aedes mosquitoes as a determinant of invasiveness and vectorial capacity by Jessica G. King, Caetano Souto-Maior, Larissa M. Sartori, Rafael Maciel-de-Freitas, and M. Gabriela M. Gomes states:
"The mosquito Ae. aegypti, primary vector of dengue, is not a natural host of Wolbachia."
"Transinfection of various strains [of Wolbachia] ... has led to field releases of wMel-transinfected Ae. aegypti in 10 countries — Australia, Brazil, Colombia, Indonesia, Sri Lanka, India, Vietnam, Kiribati, Fiji and Vanuatu — to evaluate its effectiveness in reducing dengue and other mosquito-borne diseases in human populations."
"We find that the symbiont [Wolbachia] increases the mean and the variance in Aedes aegypti susceptibility to dengue infection."
In other words: Wolbachia actually ENHANCES dengue transmission by Aedes aegypti mosquitoes.
Credit: Fair Use Portion of Variation in Wolbachia effects on Aedes mosquitoes as a determinant of invasiveness and vectorial capacity by Jessica G. King, Caetano Souto-Maior, Larissa M. Sartori, Rafael Maciel-de-Freitas and M. Gabriela M. Gomes
Credit: Screenshots of my (Rose Webster) Google Plus public posts highlighting that antibiotics worked on Zika (a virus) which supports Wolbachia as the main pathogen
Wolbachia-Induced Enhancement of Arboviral Pathogens
Unfortunately, We Need to Wait (At Least) Until This Project Ends in 2021
Credit: Fair Use Portion of Jason Rasgon's 2017 NIH Grantome
6) Perhaps the best-understood sex ratio distorting strain comes from Ostrinia moths, where Wolbachia kills males by preventing the expression of Masc, which encodes a protein early in the sex determination pathway that is required for males to develop. Even in this case, however, the mechanism by which Wolbachia has this effect remains elusive.
In vertebrates, I believe the male gonads are more vulnerable for simple reasons: they are cooler in temperature and replication of sperm is ongoing. It's an area of high cellular renewal which (I believe) Wolbachia likes to target.
Whereas, when a baby girl is born, her ovaries already contain hundreds of thousands of eggs. Location-wise, and temperature-wise, it's likely more difficult (but not impossible) for Wolbachia to infect females.
However, I would be remiss to leave out John Timmer's explanation in his brilliant 2011 post, Meet Wolbachia: the male-killing, gender-bending, gonad-eating bacteria:
"In some insects, the process of determining the visible sex is linked to the process of compensating for the fact that the two sexes generally have different numbers of chromosomes (ie, XX vs. XY). In these cases, developing as a sex that doesn't match your chromosomes can be fatal, since the gene dose is unbalanced. Wolbachia does this, too, killing off all the males."
Even Though Strong Antibiotics Fight Wolbachia Infections
If Untreated, They Could Cause Mutations That Lead to Human Cancers
Credit: Fair Use Portion of Lateral gene transfer between prokaryotes and eukaryotes by Karsten B.Sieber, Robin E.Bromley, and Julie C.Dunning Hotopp
Credit: Rose Webster (aka RoseWrites)
Why Were "Decay Rates" in an "Island Complex" Studied?
Are Aedes Aegypti Simply the Time Release Capsules For Disease?
Credit: Image Public Domain | Fair Use Portions Used From 153 Address by JFK and Fair Use Portion of Book "Deadly Cultures: Biological Weapons since 1945" by Mark Wheelis, Lajos RÃ³zsa
Demand Humans (Vertebrates) Be Tested For Wolbachia
Published November 14, 2017 by Rose Webster (aka RoseWrites on InfoBarrel)
Wolbachia-Infected Mosquito Releases are Dangerous
In April of 2017, I submitted the following concerns to the FDA:
Recommended Reading and More Ways to Help
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1) Acute Inflammatory Response, Uveal Melanoma, or Lymphoma? R/O Rickettsiales (Wolbachia)
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