Progress in Developing a Live Diagnostic Test for Chronic Wasting Disease - 1 of 2

Article Summary:

The current belief is that chronic wasting disease (CWD) and other fatal transmissible spongiform encephalopathies (TSEs) are caused by a misfolded protein called a prion. However, there is no solid evidence to support this theory.

Our laboratory has found that a spiral-shaped bacterium called Spiroplasma is present in 100% of TSE samples. This bacterium has the same biological properties as the TSE transmissible agent. We believe that Spiroplasma is the cause of CWD and other TSEs.

We are developing a diagnostic test for CWD based on this bacterium. This test will be a valuable tool for early detection and prevention of CWD.

In Jackson Hole, Good News About CWD—For Now

On December 16, 2020, wildlife officials in Grand Teton National Park received word of something they had been dreading, and for years respected disease experts said was inevitable: confirmation that an elk taken by a hunter in the park had tested positive for Chronic Wasting Disease.

State and federal wildlife managers had known it wasn’t a matter of if, but when, wapiti congregating in Jackson Hole would come down with CWD. The test result 26 months ago further corroborated fears that the always fatal disease afflicting members of the cervid (or deer family) had reached the geographic middle of the Greater Yellowstone Ecosystem.

This article was written by Todd Wilkinson. Read the rest of the article HERE.

WELCOME to the TSE RESEARCH CENTER

The PURPOSE of this website is to present information on transmissible spongiform encephalopathies (TSE) in a clear manner and to provide easy access to peer-reviewed publications for those interested in pursuing the topic in a more detailed manner. This site emphasizes the work of Dr. Frank Bastian, (Neuropathologist and Research Professor at Tulane Medical School and the University of New Orleans) with regard to the role of spiroplasma, a wall-less bacterium in the pathogenesis of TSEs.

The PROBLEM with most Government and University informational websites is that no attempt is made to critique the research reports, making it impossible for others to understand the significance of these data. There is much controversy regarding the causality of TSEs. Our research shows that prion amyloid (the TSE research focus for the past 30 years) is the result of a bacterial infection. This controversy directly reflects on Louis Pasteur's efforts to prove that replicating crystals are not the cause of infectious disease, but instead, bacteria are the culprits. Too bad we have to rehash this controversy.

The SOLUTION is to set out a new course for TSE research. Here we present data showing involvement of spiroplasma, a tiny wall-less bacterium, in the pathogenesis of TSEs. Spiroplasma are consistently associated with the TSEs, and experimental spiroplasmosis in animals shows remarkable clinical and pathological similarities to naturally occurring TSEs. A novel spiroplasma species has been isolated into cell-free media from all forms of TSE. Recent breakthrough research shows that this bacterium induces biofilm and becomes buried in a polysaccharide (complex sugar) matrix wherein the organism is resistant to physical and chemical treatments consistent with TSE.

The REWARD for our pursuing alternative research for over 30 years may be 'the TSE riddle is solvable'. Over this time there has been little progress in determining strategies for diagnosis and treatment of CJD patients because scientists have been barking up the wrong tree. If we are ever going to resolve this mystery and relieve the suffering of the many families associated with CJD cases, or the uncertainty of the health of cattle and other ruminants, then we have to go down another path of scientific investigation. This will require broader government funding to attract young researchers into TSE research.

We CONCLUDE that the scientific community must take notice of the abundant data supporting spiroplasma as a candidate causal agent of TSEs and focus on this organism for developing future strategies on handling these diseases.

Spiroplasma Presentation

 

Click HERE for a link to Dr. Bastian's latest presentation on his extensive spiroplasma research.

Data Supports Spiroplasma

It is a remarkable fact that almost all the data used to support the prion concept is conjecture. It is unfortunately the case that the wrong science is being used to address the CWD panzootic.

What follows are numerous lines of evidence supporting the role of an extreme thermo-acidophilic bacterium (spiroplasma sp.) in the pathogenesis of chronic wasting disease and the other transmissible spongiform encephalopathies (with references).

MORPHOLOGICAL

• spiroplasma identified by TEM in CJD brain tissues (Ref 1 & 2)
• spiroplasma in aqueous fluid of sheep with terminal scrapie (Ref 9) 
• experimentally spiroplasma-infected animal model (Ref 3 & 4)

RAT MODEL 

• spiroplasma induces spongiform encephalopathy (Ref 3 & 4)

DEER MODEL

• clinical evidence of brain stem lesions (Ref 8)

• morphological evidence of brain stem lesion (Ref 8)

RUMINANT MODEL- SHEEP AND GOATS

• morphological evidence of brain stem lesion (Ref 8)

• localization to eyes- retinopathy (Ref 9)

MOLECULAR DETECTION OF ORGANISM IN TSE-AFFECTED TISSUES (PCR)

• culture of spiroplasma from CWD & TSE-affected tissues (Ref 7 & 12)

• passage in embryonated eggs (Ref 7)

GROWTH IN CELL-FREE BRUCELLA SPECIAL MEDIA

• growth on agar with formation of subsurface plaques (Ref 12)

• identical biological characteristics of transmissible TSE agent as shown by enumeration of subsurface plaques on agar (Ref 13) 

IMMUNOLOGICAL

• reaction scrapie hyperimmune sera with spiroplasma fibrils (Ref 5) 

• ELISA reaction spiroplasma recombinant HSP60 with CJD sera (Ref 14) 

OTHER

• formation of alpha synuclein in spiroplasma mirum infected mammalian tissue culture

• normal prion protein isoform on cell surface is receptor for bacterium-explains interaction of prion with spiroplasma

REFERENCES

1. Bastian FO: Spiroplasma-like inclusions in Creutzfeldt-Jakob disease. Arch Pathol Lab Med l979;103:665-669.
2. Bastian FO, Hart MN, Cancilla PA: Additional evidence of Spiroplasma in Creutzfeldt-Jakob disease. Lancet l981;1(8221):660.
3. Bastian FO, Purnell DM, Tully JG: Neuropathology of spiroplasma infection in the rat brain. Amer J Pathol l984;114(3):496-514.
4. Tully JG, Bastian FO, Rose DL: Localization and Persistence of Spiroplasma in an Experimental Brain Infection in Suckling Rats. Ann Microbiol (Inst Pasteur) 84;135A:111-117.
5. Bastian FO, Jennings R, Gardner W: Antiserum to Scrapie Associated Fibril Protein Cross-Reacts with Spiroplasma mirum Fibril Proteins. J Clin Microbiol l987;25(12):2430-2431.
6. Bastian FO, Jennings R, Huff C: Neurotropic Response of Spiroplasma mirum following peripheral inoculation in the rat. Ann Microbiol (Inst Pasteur) 1987;138(6):651.
7. Bastian FO, Dash S, Garry R. Linking chronic wasting disease to scrapie by comparison of Spiroplasma mirum ribosomal DNA sequences. Exp Mol Pathol 2004;77:48-56.
8. Bastian FO, Sanders DE, Forbes WA, Hagius SD, Walker JV, Henk WG, Enright FM, Elzer PH. Spiroplasma spp. from TSE Brains or Ticks Induce Spongiform Encephalopathy in Ruminants. J Med Microbiol, 2007; 56: 1235-1242.
9. Bastian FO, Boudreaux C, Hagius SD, Sorensen S, Bulgin M, Enright FM, Elzer PH. Spiroplasma found in eyes of scrapie affected sheep. Veterinary Ophthalmology (2011) 14, 1, 10–17
10. Bastian FO, Elzer PH, Wu X (2012) Spiroplasma spp. biofilm formation is instrumental for their role in the pathogenesis of plant, insect and animal diseases. Exp Mol Pathol 93: 116-128.
11. Bastian FO. (2014) The case for involvement of spiroplasma in the pathogenesis of transmissible spongiform encephalopathies. J Neuropathol Exp Neurol 73: 104-114
12. Bastian FO, Lynch J, Hagius S, Wu X, McCormick G, Luther DG, Elzer PH.(2018) Novel Spiroplasma Spp. Cultured From Brains and Lymph NodesFrom Ruminants Affected With Transmissible Spongiform Encephalopathy. J Neuropathology & Experimental Neurology, 77: 64-73.
13. Bastian FO, Lynch J, Wang W-H. Novel Spiroplasma sp. Isolated From CWD Is an Extreme Bacterial Thermoacidophile That Survives Autoclaving, Boiling, Formalin Treatment, and Significant Gamma Irradiation. J Neuropath Exp Neurol 2019 IN PRESS
14. US 7,888,039 B2 Feb. 15, 2011

 

A Historical Note

Dr. Bastian has been researching the pathogenesis of the transmissible spongiform encephalopathies (TSE) for over four decades. He grew up in Saskatchewan, Canada (a current hotbed of the ongoing panzootic of chronic wasting disease (CWD), a rapidly spreading TSE in deer, moose, and elk populations. CWD was discovered to have spread from Fort Collins, Colorado to currently involve 29 States and four Canadian provinces). CWD has also a global presence involving deer populations in Europe and recently found in reindeer herds in Sweden and Norway. CWD is also affecting the cervid population in South Korea. Dr. Bastian is a Neuropathologist trained under Professor Steven Vogel at Duke Medical School in Durham NC (1968-1972).

In the 1960s the transmissible agent of scrapie (a TSE known for over 300 years in sheep) would not pass through a 100 nanometers filter (the size of a large virus thus the idea of a slow virus disease- a Herpes virus measures 140 nanometers in diameter and is easily visualized by Electron Microscopy [EM]).

After finishing his training at Duke, and a productive fellowship with Alan Rabson at the National Institutes of Health (NIH) in Bethesda MD, involving studies of latency of Herpes viruses in brain and spinal cord, he assumed a position of Assistant Professor of Pathology and Neuropathology at Baylor College of Medicine in Houston, TX. At this juncture, Dr. Bastian diagnosed a case of Creutzfeldt Jakob disease (CJD) which is a fatal TSE in humans by brain biopsy. Since the brain tissues in the brain biopsy were ideally prepared for ultrastructural study, he used his expertise in EM to search for morphological evidence of the TSE transmissible agent, which as mentioned above was virus-like in size. After extensive search, he found a helical bacterium in the CJD-affected brain biopsy tissues that closely resembled a group of phytopathogens (Spiroplasma spp.) recently described by Dr. Joseph Tully, Head of the Mycoplasmology Section at the NIH.

Subsequently Dr. Bastian moved to University of Maryland in Baltimore, MD and set up a close relationship with Dr. Tully at the nearby NIH to pursue his quest and study the biology of these novel organisms. He set up a Spiroplasma laboratory at University of South Alabama as Professor and Director of Neuropathology. During his tenure at USA he established a link between spiroplasma and the TSEs using molecular studies including PCR. He published a comprehensive book on CJD and the other TSEs while on faculty. Dr. Bastian moved his laboratory after Katrina to Department of Veterinary Science at the LSU Agriculture Center where he conducted experiments in small ruminants wherein he completed Koch’s postulates regarding proof of concept that a novel spiroplasma was the cause of TSEs. He then had a research breakthrough in his laboratory wherein he was able to isolate the novel spiroplasma from TSE-affected tissues including CJD, scrapie and CWD into cell-free media and as colonies on agar plates. He showed that the spiroplasma TSE isolates were resistant to boiling, radiation and formalin fixation (an extreme thermoacidophilic bacterium) consistent with the known biological properties of the TSE transmissible agent. All these studies are recorded in peer-reviewed publications. His laboratory is currently established in the AMRI business complex on the University of New Orleans (UNO) campus. Dr. Bastian is currently Research Professor at UNO and Tulane Medical School.

Interview with an expert

Spiroplasma may cause Creutzfeldt-Jakob Disease.  An interview with a leading expert in infectious diseases: Dr. Frank Bastian.

What is Creutzfeldt-Jakob Disease (CJD?)

CJD is a transmissible disease characterized by spongy degeneration of the brain. It strikes about one in 1 million people annually. About 5% of CJD cases occur in families or among certain ethnic groups. The disease has been transmitted inadvertently during medical treatment and surgical procedures. CJD has been transmitted by the administration of growth hormone derived from the pituitary glands of people who died from CJD. The disease also has been transmitted by transplantation of corneas and dura tissue obtained from people who died from CJD. CJD has been transmitted experimentally to monkeys by oral feeding of contaminated tissue and to lab animals by infusion of white cells.

CJD symptoms usually appear when people are in their 40s, 50s, and 60s. Ninety percent of people with CJD die within a year of the onset of signs and symptoms. Most die in four to six months.

CJD is difficult to diagnose early. At first, people experience psychological disturbances and bizarre behavior, and then proceed to dementia, memory loss, stiffening of muscles, and difficulty walking. Although this illness may be confused with Alzheimer's disease, people with Alzheimer's usually do not develop neurologic signs. The clinical diagnosis is usually in error at least 25% to 30% of the time.

Researchers suspect that 10 people in Britain who contracted a CJD-like illness were infected by cows afflicted with bovine spongiform encephalopathy. How do these British cases compare with classic CJD?

Researchers showed that the CJD-like illness in Britain had a different clinical course than classic CJD. All of the British cases occurred in people younger than 42 years; in fact, three cases occurred in teenagers. Moreover, the duration of illness exceeded a year in five of the 10 cases. The CJD-illness also had a pathology different from CJD. For example, the CJD variant is characterized by extensive amyloid plaque formation and by degeneration of the cerebellum.

You postulate that Spiroplasma bacteria causes CJD. Please describe them.

Spiroplasmas are similar to mycoplasmas in that they do not have a cell wall and have among the smallest genomes of any living organisms. Spiroplasmas, which were only discovered in 1976, are present in the hemolymph of almost all insects. There probably are several million strains of spiroplasmas. They are very fastidious organisms. Many spiroplasmas grow at fairly low temperatures. A few strains grow at 37° C, human body temperature. Many spiroplasmas cause diseases in plants. Spiroplasmas usually are associated with a vector. For example, a leaf hopper carries a spiroplasma that infects orange trees. These organisms are essentially resistant to all bacteriocidal antibiotics.

How did you connect spiroplasmas with CJD?

In 1976, I examined a brain biopsy from a patient with CJD using electron microscopy. I saw a spiral structure foreign to the tissue. It had features of the newly reported spiroplasmas. In 1981, a team in New York reported finding a fibril protein in scrapie-infected brain tissue. This scrapie-associated fibril (SAF) protein was 4 nm in diameter and 200 nm long. In 1983, the team looked at various tissues of CJD and kuru and demonstrated scrapie-associated fibrils consistently in these diseases but not in control tissues. These SAF are identical morphologically to the internal fibrils of spiroplasmas. This similarity, in my mind, as a pathologist, solidifies the link between spiroplasmas and CJD.

Moreover, antibodies to SAF react with internal fibrillar proteins from Spiroplasma and digested brain material from people with CJD, suggesting that these proteins essentially are the same. I have shown in my laboratory that spiroplasmas are neurotropic. If you inoculate them peripherally into suckling rats, they will eventually localize to the brain tissues. The organisms will produce a persistent infection and produce a spongy change in the brain tissue of these animals. The neuropathologic changes are similar to those seen in CJD.

Another piece of circumstantial evidence is that spiroplasmas are within the size range of the agent that transmits CJD and other transmissible spongiform encephalopathies. Spiroplasmas will pass through a 50 nm-pore filter. The transmissible agent's size has been determined to be 42 nm.

What would prove that Spiroplasma causes CJD and related illnesses?

Definitive evidence of this link would be to demonstrate that spiroplasma DNA occurs in the brain tissue of people with CJD and related illnesses, but not in brain tissue from controls. This could be done by using polymerase chain reaction assays to detect nucleotide sequences in genes unique to spiroplasmas and common to all types of spiroplasmas. I have 15 types of spiroplasmas in my laboratory and am in the process of looking for nucleotide sequences common to all of them.

Why have investigators failed to find a causative agent despite 30 years of looking?

The obvious way to look for an agent directly is by electron microscopy, but this method may not be appropriate for spiroplasmas. Spiroplasmas are similar to mycoplasmas, and it is a well-known phenomenon that mycoplasmas are able to blend with cell membranes. What happens, possibly, is that spiroplasmas essentially fuse with host-cell organelle membranes, thereby blending with the background, so you would not see it unless you had a marker to label it. Developing such as marker has been difficult because spiroplasmas are difficult to cultivate. No more than half of the known strains are culturable.

After inoculating spiroplasma in suckling rats, we examined brain tissues by electron microscopy early in the infection and could document the organisms in the tissues. They appeared as membrane-bound forms, except for the one instance in which I observed the spiral form. Later in infection, when we knew that the tissues were infectious by broth culture, we couldn't find any evidence of the organism by looking at the tissues extensively with electron microscopy.

What do you make of the prion theory?

The prion is a red herring. Prions are thought to be self-replicating proteins. Some researchers believe prions are the cause of CJD and related illnesses because they have found prions in brain tissue from people with CJD and sheep with scrapie but not in normal brain tissue. A shortcoming in the prion theory is that CJD and scrapie can be transmitted without prions. Brain material from which the prion has been removed with antibodies can still infect animals. Moreover, the prion has been found in unrelated disease processes, such as Kawsaski syndrome and inclusion body myositis.

The protein that some researchers believe causes CJD and related illnesses is a reconfigured normal host protein. Studies have shown that if you take the gene for the normal protein out of the mouse, the mouse does not develop spongiform encephalopathy when injected with infectious material. The protein is likely important in the pathogenesis of the disease. Researchers have jumped to conclusions from that evidence to suggest that the protein has to be the causative agent, and they have not considered any other possibility. It is quite possible that spiroplasmas may be inducing the formation of this prion protein to protect itself from the immune system.

The immune system is very important in the pathogenesis of CJD. We know that the agent replicates in the spleen and lymph nodes and occasionally causes an immunologic reaction. Auto-antibodies are characteristically seen in the late stages of experimental and naturally occurring disease.

A common phenomenon among the mycoplasmas is that the organisms bind host proteins that often are of identical molecular weight to their surface proteins and, therefore, are looked at by the immune system as being the same as the host. The spiralin protein on the surface of spiroplasmas shows a migration pattern on gel electrophoresis with a molecular weight of 27,000 Da to 30,000 Da, similar to that of the so-called prion protein. This biochemical similarity is compatible with spiroplasma etiology.

If spiroplasmas turn out to be the cause of CJD and related illnesses, could antibiotics be used to treat them?

Spiroplasmas are susceptible to erythromycin and tetracycline, but these drugs would not be of value in treating CJD or related illnesses because the drugs are bacteriostatic, not bacteriocidal. I would hope that one of the newer quinolones would be useful in killing the organism.

Why might humans have contracted a CJD variant from cows with bovine spongiform encephalopathy (BSE)?

The transmission from cows to humans might be related to an increase in virulence in the BSE agent. It is known that a pathogen's virulence can be increased by passing a strain of the agent through a series of hosts of the same species. Experiments with scrapie have shown that such serial passage of the scrapie agent can increase the virulence and cut the incubation time in half.

In Britain, sheep and cattle tissue used to be ground up and added to cattle feed. Cows may have initially contracted BSE by eating feed containing scrapie-infected tissue. Eventually, infected cattle tissue was fed to cattle, so the infectious agent was passed through one cow to another. This may have increased the virulence of the infectious agent to the point where it could infect humans.

Spiroplasma and Transmissible Spongiform Encephalopathies

 by Ed Gehrman

Transmissible Spongiform Encephalopathy (TSE) is identified by the plaques of mutated amyloid protein that form within the brain tissue and destroy synapses and neurotransmitter functions and take on a characteristic sponge or Swiss cheese appearance. CJD, Scrapie, and Kuru are all members of this degenerative disease family, afflictions known about for over two hundred years but not studied intently until the early sixties when they were found to be transmissible.

Dr. Carleton Gajdusek was a young researcher looking for unusual diseases when he visited the Fore Peoples of Papau, New Guinea during the late 1950s. The Papuans of those years were suffering from a population density that put a strain on very limited resources. They practiced female infanticide and cannibalism and were in a constant state of warfare with their neighbors over land and pigs. Severe limitations on normal heterosexual relations were imposed; the men spent most of their time at the men's clubhouse where they prepared for war and engaged in homosexual relations with the young boys. This homosexual activity was all part of an elaborate bonding thought needed to ensure macho warriors and dependable compatriots. The warfare was brutal, often hand-to-hand; capture meant being tortured and killed. Solidarity was essential and achieved through the sharing of semen. The females of the group were disrespected and often abused because they were thought to steal the men's strength and resolve in battle as well as their vital semen. The malnourishment of females and young children was part of this intense process; they supplemented their diet by eating anything that "crawled or crept". Midwives ate placentas of the newborn and women dug up the partly decomposed bodies of relatives and ate and shared with young children the flesh, brains, and the accumulations of maggots and mites. This was not a religious ceremony but an attempt to fend off malnutrition. (1)

Gajdusek observed that some of the Fore women and a few children died from symptoms indicating a neurological disorder: dementia, frenzied behavior, blindness, and eventual agonizing death. He studied the tribal dynamics and soon hypothesized that the condition, known as Kuru, came from their habit of eating the brains of dead relatives; he brought some diseased brain tissue back to the USA. Gajdusek soon discovered that when he made a broth from the Kuru tissue and injected this mixture into lab animals, they too exhibited the Kuru symptoms. He then processed Kuru diseased lab animals' brains and injected the mixture into other lab animals. They also died the same excruciating deaths. This meant that the condition could be transmitted from organism to organism and was therefore transmissible, hence Transmissible Spongiform Encephalopathy (TSE).

Gajdusek and his colleagues at the National Institute of Health were never able to isolate or positively identify the agent that causes the TSE even though they've been trying since the early sixties. Scrapie and CJD were also studied and found to be transmissible. All this was well-known underground medical information; many doctors refused to autopsy CJD victims. For years the NIH conjectured that the infective culprit was a "slow virus". Nothing seemed to destroy the agent; not heat, cold, or any of the normal chemical disinfectants. Nor could they find a trace of its chemical or molecular identity. Furthermore, the virus didn't cause inflammation so antibodies failed to leave a calling card. Some completely new agent was essential.

Another tenacious TSE researcher is Dr. Frank O. Bastian, MD, a professor of pathology and director of neuropathology at the University of South Alabama, Mobile. He has published numerous research articles relating to the etiology of Creutzfeldt-Jakob Disease and also edited a book entitled Creutzfeldt-Jakob Disease and Other Transmissible Spongiform Encephalopathies. (2)

In 1976, Bastian examined a brain biopsy from a patient with CJD using electron microscopy. He saw a spiral structure foreign to the tissue. It had features of the newly reported spiroplasmas (spiroplasmas were only discovered in 1976). In 1981, a team in New York reported finding a fibril protein in scrapie-infected brain tissue. This scrapie-associated fibril (SAF) protein was 4 nm in diameter and 200 nm long. In 1983, the team looked at various tissues of CJD and Kuru and demonstrated scrapie-associated fibrils consistently in these diseases but not in control tissues. These SAF were identical morphologically to the internal fibrils of spiroplasmas.

Moreover, antibodies to SAF react with internal fibrillar proteins from Spiroplasma and digested brain material from people with CJD, suggesting that these proteins essentially are the same. This similarity solidified in Bastian's mind the link between spiroplasmas and CJD.

Dr. Bastian has postulated that Spiroplasma bacteria causes CJD and other TSE. His twenty years of research indicates a role for Spiroplasma. The evidence includes the following: spiroplasma-like inclusions were seen in brain biopsies from patients with CJD (Arch Pathol Lab Med. 1979;103:665-669); spiroplasma internal fibril proteins are identical morphologically to those seen in TSE's; the spiroplasma proteins show immunological cross-reactivity with the TSE proteins (J Clin Biol. 1987;25:2430-2431); and spiroplasma, when inoculated into rodents, produces a similar neuropathology (Amer J Pathol. 1984;114:496-514). Spiroplasmas are present in the hemolymph of almost all insects; there probably are several million strains. They can also cause diseases in plants but are usually associated with a vector. For example, a leaf hopper carries a spiroplasma that infects orange trees Spiroplasmas are similar to mycoplasmas. They do not have a cell wall (cell wall deficient) and have among the smallest genomes of any living organism. Mycoplasma are the smallest and perhaps the oldest life form. These bacteria, one cause of "walking pneumonia", are thought by many to be rather fragile, but nothing could be further from the truth. They tolerate extreme fluctuations in temperature, lay dormant in the soil for generations and survive the harshest elements; only drano-like chemicals kill them effectively outside the body. Under normal circumstances, our immune system efficiently deals with this complicated, membrane-enclosed piece of DNA. A common phenomenon among the mycoplasmas is that the organisms bind host proteins that often are of identical molecular weight to their surface proteins and, therefore, are looked at by the immune system as being the same as the host. The spiralin protein on the surface of spiroplasmas shows a migration pattern on gel electrophoresis with a molecular weight of 27,000 Da to 30,000 Da, similar to that of the prion protein. This biochemical similarity is compatible with spiroplasma etiology.
Bastian was able to show that spiroplasmas were neurotropic. When inoculated peripherally into suckling rats, they will eventually localize to the brain tissues. The organisms will produce a persistent infection and produce a spongy change in the brain tissue of these animals. The neuropathologic changes are similar to those seen in CJD. Spiroplasmas are also within the size range of the agent that transmits CJD and other transmissible Spongiform encephalopathies. Spiroplasmas will pass through a 50 nm-pore filter. The transmissible agent's size has been determined to be 42 nm.

The obvious way to look for an agent directly is by electron microscopy, but this method may not be appropriate for spiroplasmas. Spiroplasmas are similar to mycoplasmas, and it is a well-known phenomenon that mycoplasmas are able to blend with cell membranes. What happens, possibly, is that spiroplasmas essentially fuse with host-cell organelle membranes, thereby blending with the background, so you would not see it unless you had a marker to label it. Developing such a marker has been difficult because spiroplasmas are very fastidious (difficult to cultivate)organisms. 

Bastian also inoculated suckling rats with spiroplasmas, and examined their brain tissues by electron microscopy early in the infection process; he documented the organisms in the tissues. They appeared as membrane-bound forms, except for the one instance where he observed the spiral form. Later in infection, when he knew that the tissues were infectious by broth culture, he couldn't find any evidence of spiroplasmas by looking at the tissues extensively with electron microscopy. Bastian insists that the infection-related protein that most researchers refer to as a "prion" is produced by the host in response to the infection and is not the causative agent. Prions are thought to be self-replicating proteins. Some researchers believe prions are the cause of CJD and related illnesses because they have found prions in brain tissue from people with CJD and sheep with scrapie but not in normal brain tissue. Bastian states that a shortcoming in the prion theory is that CJD and scrapie can be transmitted without prions.
Brain material from which the prion has been removed with antibodies can still infect animals. Moreover, the prion has been found in unrelated disease processes, such as Kawsaski syndrome and inclusion body myositis. Prion researchers have jumped to conclusions and have not considered any other possibility. It is quite possible that spiroplasmas may be inducing the formation of the prion protein to protect itself from the immune system. The immune system is very important in the pathogenesis of CJD. The agent replicates in the spleen and lymph nodes and occasionally causes an immunologic reaction. Auto-antibodies are characteristically seen in the late stages of experimental and naturally occurring disease. The gene for the host protein is located on the chromosome in the region of the major histocompatibility complex (MHC) in the mouse. "Occasionally, you see elevation of immunoglobulins; there are morphological alterations of the leukocytes; there is leukopenia," Bastian explains, "and auto antibodies are characteristically seen in the late stages of both experimental and naturally occurring infection. There is partial MHC restriction in both human and animal disease."

"The immune reaction seen in these Spongiform diseases can be explained by superantigen activity, Bastian says. He notes that, normally, an antigen is presented to the cell surface in the MHC and interacts with the T-cell receptor—the antigen lying in a groove in the T-cell-MHC sets in motion the standard reaction. A superantigen, on the other hand, binds outside the groove of the T-cell and interacts with the MHC. This results in some immunoglobulin production, but only transiently. The major effect is clonal deletion of T cells, resulting in a state of immune tolerance. Autoantibodies can also form. In Spongiform diseases, PrP presumably acts as a superantigen. It is noteworthy that inclusion body myositis, a condition in which prions are seen, is an established super antigen disease."

Dr. Bastian also notes that investigators have reported transmitting a TSE to mice from hay mites gathered from farms in Iceland where scrapie is endemic (Lancet. 1996;347:1114). He is virtually certain that these hay mites contain spiroplasma, noting that the investigators have not so far found PrP in the mites.
If hay mites can cause TSE, why couldn't the same be true for the maggots and mites on the Fore corpses? (3) Could Gajdusek have overlooked the main factor connecting Kuru to the Fore women and children? Was the initial cause of Kuru the ingesting of large quantities of maggots and mites by protein-famished women and children? We know that the maggots and mites contain spiroplasma. By eating the brains of the Papuans that died from Kuru, the disease (Spiroplasma) was retransmitted to those remaining, in a deadly cycle. Transmissible Spongiform Encephalopathies will continue to be misunderstood unless we begin to study and understand these simple connections.

ENDNOTES

(1) This information comes from several sources. It is well known to anthropologists that these conditions existed among the Fore peoples and many other New Guinea tribes like the Sambia. I know it's hard to believe in these modern times but we must if we are to understand the world in which we live. My main source is Our Kind by Marvin Harris; Harper & Row; 1989. He took much of his information from Shirley Lindenbaum, Kuru Sorcery: Disease and Danger in the New Guinea Highlands; 1979; Mayfield

(2) JAMA August 14, 1996 DC Capital Conference spring 1996 A dissenting view on the cause of Mad Cow Disease Bastian regards the prion theory as a red herring. The cause of Transmissible Spongiform Encephalopathies (TSEs), he says, is a conventional microorganism -- a mollicute or, more specifically, a spiroplasma. "The infection-related protein is produced by the host in response to the infection,". 

[Infectious Disease News Homepage] (June 1996) Spiroplasma may cause Creutzfeldt-Jakob Disease - An interview with a leading expert in infectious diseases, Frank O. Bastian, MD. In 1992, Bastian arranged an international symposium on Bovine Spongiform Encephalopathy.
I used information, quotes, and descriptions from the above article and interview to weave together Dr Bastian's ideas, knowledge and words, with my own research. I edited and rearranged both words and sequences for coherence's sake. I did the best I could to convey this important message.
I've had three phone conversations with Dr. Bastian. He was cooperative and helpful at first and sent me much useful information which I have included. But he cut a scheduled interview short when I began to suggest that biowarfare research might help to spread the TSE agent, inadvertently. I called one more time and he refused to talk. He has refused to answer a long letter I wrote. I thought it both rude and arrogant, but even with that nonsense, I still believe Bastian's elegant hypothesis is far more rational than any I've studied.

(3) Common arthropods occurring on dead bodies: The Acari, or mites as they also are called, are small organisms, usually less than a mm in length. Mites occur under the dead body in the soil, during the later stages of decay. Many mites are transported to the body via other insects, such as flies or beetles. Other mites are soil-dwelling forms that can be predators, fungus feeders, or detritus feeders. Most species will be found in soil samples from the seepage areas under the body. Sarchophagidae Among the Sarcophagids we find the large flesh-flies with red eyes and a grey-checkered abdomen. These flies do not deposit eggs, but larvae on the corpse. They are, together with the Calliphorids, among the first insects to arrive at the corpse. The larvae are predators of blowfly larvae, as well as carrion feeders. Many Sarcophagids are feeding on snails and earthworms.

NOTE: The original article appeared in the Sonoma County Free Press, which is archived HERE.