April to June 2009 in Tahiti, French Polynesia
Wolbachia-Infected Aedes Began to Infect Our Food Chain But This Technology Has Been "In Development" For Over 30 Years
Credit: Alf Oldman on flickr [Public Domain] Taken on March 23rd, 2017
The WHO has been working on Dengue for decades
The 2011 paper by Chambers et al. confirmed that between April and June 2009 on the island of Tahiti, French Polynesia, semi-field experiments using Wolbachia-infected male Aedes mosquitoes were conducted.
The sites were ‘motu’ islands, selected due to their small size, isolation and absence of human inhabitants.
N.B: Absence of human inhabitants.
As I stressed in my post Wolbachia-Infected Mosquitoes Might Reduce Dengue, Enhance Zika, and Cause a Million Souls to Become Sterile:
The study Assessing key safety concerns of a Wolbachia-based strategy to control dengue transmission by Aedes mosquitoes states:
"The possible horizontal transfer to mosquito predators and non-predator species or environments in the vicinity of the mosquitoes were evaluated."
But they only evaluated: spiders, spider eggs, soil samples, plant leaves, plant roots, earthworms, and millipedes (in Australia).
Shockingly, the authors concluded:
"All together, the results of these experiments show that there is no transmission of the bacteria to any of the environmental samples studied."
Zika Became Devastating
in French Polynesia
Since Wolbachia is co-inherited with mitochondria, natural selection acting over the bacterium will also affect mitochondria.
Depending on the infection context, this hitchhiking effect may increase or decrease mitochondrial genetic diversity (Keller et al., 2004, Dean et al., 2003 and Shoemaker et al., 2003).
Between October 2013 and April 2014, French Polynesia experienced the largest Zika virus outbreak ever recorded in the country.
During this period of time, 32,000 patients were assessed for the infection. And there was a 20-fold increase in incidence of GBS (Guillain-Barré syndrome) in French Polynesia compared with the previous four years.
"Wolbachia could invade from a very low prevalence" (Dean JL, Dobson SL (2004); Plichart C, Legrand AM (2005).
Examples of neutral or pro-pathogenic effect of Wolbachia include: Brugia pahangi (Dutton and Sinkins, 2005); Japanese encephalitis (JE) Virus (Tsai et al., 2006); Drosophila C Virus (Osborne et al., 2009); and Plasmodium gallinaceum (Baton et al., 2013).
Wolbachia Enhances West Nile Virus (WNV) Infection in the Mosquito Culex tarsalis.
Five independent research teams have determined Culex can be Zika vectors. This study, which received no press, Zika Virus in Salivary Glands of Five Different Species of Wild-Caught Mosquitoes from Mexico clearly states:
"Here, we report the isolation in cell culture of ZIKV from different body parts of wild-caught female mosquitoes (Ae. aegypti, Ae. vexans, Culex quinquefasciatus, Cx. coronator, and Cx. tarsalis) and whole male mosquitoes (Ae. aegypti and Cx. quinquefasciatus) in Mexico."
Whole male mosquitoes is worrisome, as this indicates that Zika is maternally inherited (just like Wolbachia).
And there already is study literature that suggests Wolbachia infects humans. The 2015 study Detection of Wolbachia genes in a patient with non-Hodgkin's lymphoma states:
"Wolbachia spp. should be further evaluated as causes of human infection, especially as Wolbachia infection of mosquitoes is increasingly considered to be a tool for interfering with mosquito-borne transmission of human pathogens."
NB: The filariodea coxI gene was not found in this case (which points to direct mosquito-to-human transmission).
Dr. Ayres Warned the World About Culex Feb. 2016
Others confirmed the same; but the media ignores it
Credit: Rose Webster (aka RoseWrites)
The Case for Zika Acting in Tandem With Wolbachia
The latest findings support it
On October 10th, 2017, Dr. Edison Luiz Durigon, confirmed the presence of "compartmentalized strains" of Zika. Notable quotes include:
"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."
"... the male patients continued to excrete large amounts of Zika virus in their semen for up to six months. One was found to have the virus in his saliva for three months."
"Zika continued to replicate in the patient’s testicular cells all this time, and using an electron microscope, we could see that the spermatozoa were formed already infected ... This means a conception could occur with infected sperm."
The Serine-to-Asparagine Substitution
Could this be the smoking gun?
On October 12th, 2017, the Chinese Academy of Sciences published Zika was a Mild Bug, a New Discovery Shows How It Turned Monstrous. Quotes which point to Wolbachia-infected Aedes releases:
"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."
The group of 24 scientists "slowly modified the older strains, making one change to a protein at a time ... After trying seven different amino acids, they found the culprit. It was a single amino acid called serine that, when replaced by another amino acid called asparagine, caused the devastating effect."
As noted in my post Tracing Zika's Path to Florida, Culex, and Wolbachia
, according to the study Decreased Diversity but Increased Substitution Rate in Host mtDNA as a Consequence of Wolbachia Endosymbiont Infection
by D. DeWayne Shoemaker, Kelly A. Dyer, Mike Ahrens, Kevin McAbee and John Jaenike:
"Wolbachia, and probably other maternally transmitted endosymbionts, can severely depress levels of mtDNA diversity within an infected host species. In contrast, such infections may increase the rate of substitution in mtDNA."
The 2013 study Characterization of a serine-to-asparagine substitution at position 123 in the Japanese encephalitis virus E protein confirms:
"Amino acid position 123 in the E protein of Japanese encephalitis virus (JEV) determines viral growth properties and pathogenicity. The majority of JEV strains have a serine residue at this position (E(123S)); however, JEV with an asparagine residue (E(123N)) has also been isolated."
On page 26 of the 175-page PDF titled Emerging Infectious Diseases (Vol. 11, No. 8, Aug 2005) provides additional evidence that serine-to-asparagine substitutions are more pathogenic:
"Additionally, all 26 Nepal sequences exhibited a serine-to-asparagine substitution at position 189 (S189N) that had also been observed in the 6 isolates from Germany, as well as in a few isolates from Asia characterized at the end of the 2003–2004 influenza season."
And the 2011 paper Effect of an Asparagine-to-Serine Mutation at Position 294 in Neuraminidase on the Pathogenicity of Highly Pathogenic H5N1 Influenza A Virus confirms the same:
"Importantly, VN1203 virus possessing the NA N294S substitution was a potential killer of ferrets, underscoring the importance of monitoring the emergence of the NA N294S substitution in circulating H5N1 viruses."
And the authors made these recommendations:
- At present, several NA inhibitors are available, such as laninamivir and peramivir.
- Laninamivir (aka CS-8958), is effective against oseltamivir-resistant viruses. But, this inhaled drug is not appropriate for severely ill or infant patients.
- In contrast, peramivir is an intravenous NA inhibitor that can be given to patients who cannot be treated with inhaled drugs; however, some oseltamivir-resistant viruses are cross-resistant to peramivir.
- To minimize the likelihood of emergence of NA inhibitor-resistant viruses and effectively treat patients infected with such viruses, we should consider stockpiling a variety of NA inhibitors.
Zika is MORE Related to Culex-Associated Viruses
Almost 1/2 of Bulbuls Tested had Zika (Okia et al. 1971)
Credit: ©2017 Rose Webster (aka RoseWrites) All rights reserved
ZIKV Genomes Must Cover UTRs [untranslated regions]
Yet those submitted to Genbank have a protein annotation that differs SIGNIFICANTLY
On September 7th, 2017, Theys et al. found gross errors with the genomic Zika sequences (e.g., GAN KU940224). These were submitted to Genbank with a protein annotation that differs significantly from the curated NCBI reference genome. The authors noted:
"Complete ZIKV genomes must cover UTRs and should be indicated accordingly when, in fact, limited to the complete CDS (with partial UTRs) or containing large regions of undetermined nucleotides."
Genetic Engineers Often Introduce Conservative Mutations
But serine-to-asparagine substitutions are NOT one of them
Conservative mutations (aka directed mutagenesis) is used to study the role of certain amino acids. For example, replacement of a serine or threonine by an aspartate or glutamate introduces a negative charge that mimics the effect of phosphorylation of these residues.
Some missense mutations are said to be "conservative" as the changed amino acid residue has a similar property to the original residue.
For example an aspartic acid residue might be replaced by a glutamic acid residue without altering the function. According to Chapter 9 of When a different amino acid will do: conservative mutations:
- threonine may replace serine
- asparagine may replace glutamine
- arginine may replace lysine
Horizontal Gene Transfers Occur Between Different Species
A black-widow venom gene ended up in a virus that infects Wolbachia
Credit: Rose Webster (aka RoseWrites)
One Phage (Zika?) Could Modify Wolbachia Strains
And there's evidence that an entire segment was deleted as a single event
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 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)."
And according to this ResearchGate post:
"PCR-based screens (discussed later) for the presence/absence of single copy genes within this region indicate that the entire segment was deleted as a single event, rather than through gradual genomic erosion."
Drosophila melanogaster is used to study the human brain
Eliminate Dengue confirmed the Wolbachia stains they use:
Credit: Rose Webster (aka RoseWrites)
After 30 Years of Development ... Source: The Atlantic
Recent reports state "about a decade" but that is a lie. Here's proof:
Credit: Fair Use Portions of Time and The Atlantic Confirming Wolbachia-Infected Aedes Worked on Over 20 to 30 Years
The Wolbachia-Infected Aedes Patent Confirms
the replacement of an aspartic acid (D) for asparagine (N)
I invite scientists (without a conflict of interest) to consider these exact quotes from the patent titled Modified arthropod and method of use US 20110145939 A1:
FIG. 47. Protein sequence alignment of the WD0200 proteins of wMelPop and wMelPop-CLA, showing the mutation of one aspartic residue (D) into asparagine (N).
During the adaptation of wMelPop to mosquito cell culture in our laboratory, the sequence of this gene has mutated resulting in the substitution of a C residue for a T in wMelPop-CLA.
This nucleotide change results in the replacement of an aspartic acid (D) for asparagine (N) in the C-terminus of the encoded protein.
How Long Can We Ignore Wolbachia-Infected Aedes?
Credit: All Images: Public Domain | Text by Rose Webster (aka RoseWrites) All Rights Reserved
Over 6 Years Later, 90 Percent of Mosquitoes Still Infected
It will take decades to restore the ecological balance in some regions.
Credit: Fair Use Portions of MosquitoMate's FAQ and 90 Percent Still Infected Over Six Years
My FDA Comment RE: Wolbachia-Infected Mosquito Releases are Dangerous
By Rose Webster | Published on Apr 23rd, 2017