Was Polio Ever Properly Purified and Isolated?: Part One

In order to properly purify a “virus,”, the particles must be freed from anything that can contaminate, debase, adulterate, and/or pollute it. To be properly isolated, the particles must be separated from everything else that could potentially be contained within the sample. Without doing this, there would be no certainty that the particles from a mixed sample chosen and assumed to be a “virus” truly are one. There would be no way of knowing if any of the other substances contained within the sample were in fact the true culprit causing disease. This is why it is absolutely imperative that the particles assumed to be a “virus” come directly from a sick human to be properly purified/isolated first instead of being subjected to either tissue or cell cultures where the unpurified sample will become even more so with added animal DNA, antibiotics/antifungals, fetal bovine serum, chemicals, nutrients, etc.

It’s fair to ask if this process has ever been properly carried out for any alleged “virus,” and to date, I have not found a single original “virus” paper ever having done so. What is seen, however, are claims of purification/isolation in the titles of the scientific papers and mainstream headlines. However, do these proclamations of purifcation/isolation actually hold up when the evidence is examined?

Take, for instance, Polio. When discussing Polio, there are a few names that regularly come up in the conversation about who were involved in the discovery process. You will most likely hear about Karl Landsteiner and Erwin Popper as the ones who originally “discovered” the “virus” in 1908 through grotesque experiments on monkeys. You will also hear about Charles Armstrong’s research with the Lansing strain in 1939 which was the precursor to John Franklin Enders “successful” cultivation of the strain in nonnervous tissue cultures in 1949. This apparently paved the way for Jonas Salk to create the Polio vaccine in 1953 as well as Albert Sabin’s oral vaccine in 1961.

Two people you most likely won’t hear about unless you dig deeper are Hubert Loring and Carlton Schwerdt, which is odd considering these two are given credit for the purification/isolation of Polio in 1946-1947. Loring and Schwerdt should be at the top of every Polio conversation yet they seem to be consistently left out of the Polio discovery beyond some vague mentions here and there. For example, the Polio timeline from the History of Vaccines site:

Or this snippet from the Britannica Polio timeline:

Polio through history

“The poliovirus itself was discovered in 1908 by a team led by Viennese immunologist and future Nobel Prize winner Karl Landsteiner. The existence of telltale antibodies specific to the virus circulating in the blood of infected persons was discovered only two years later. In 1931 two Australian researchers, Frank Macfarlane Burnet and Jean Macnamara, using immunologic techniques, were able to identify the different serotypes of the poliovirus. (Burnet was to receive a Nobel Prize in 1960.) In 1948 the team of John Enders, Thomas Weller, and Frederick Robbins, working at Harvard Medical School in Massachusetts, showed how the virus could be grown in large amounts in tissue culture (an advance for which they shared a Nobel Prize in 1954). From there it was only a short step to an announcement in 1953 by Jonas Salk at the University of Pittsburgh, Pennsylvania, that he had developed an effective killed-virus vaccine.”

https://www.britannica.com/science/polio/Polio-through-history

Not a single mention of Loring and Schwerdt nor the supposed purification/isolation of Polio. One would think this would be a highly regarded moment in history. What could be the reason for leaving this important information out of the Polio picture?

Needless to say, trying to find out much about the methods of their work or even digging up some of their publications was a challenging task. Fortunately, I was able to gather up some information from a few sources and studies. Presented below is what I could find on the supposed purification/isolation of Polio.

From an article posted in January 2021:

Loring and Schwerdt, and their search for poliovirus

“Stanford scientists Hubert Scott Loring and Carlton Everett Schwerdt announced their successful isolation of poliovirus on January 10, 1947. Even though it was only 80% pure and extracted from cotton rats infected with polio, it was a breakthrough at the time and a giant stride in our continuing fight against polio.”

“Loring’s laboratory was characterised by a friendly atmosphere and subdued excitement. With his students, he was involved in two major areas during this time – the purification of the poliomyelitis virus and the structure and metabolism of ribonucleic acids.

Along with his student Schwerdt, Loring spent three years searching for the poliovirus. Their efforts led to the successful isolation of the Lansing strain of the poliovirus in 1946. Schwerdt completed his Ph.D. in biochemistry by the time their results were announced on January 10, 1947.

Tempers excitement

Loring and Schwerdt were able to obtain the virus with at least 80% purity. They were able to extract it from cotton rats, the only species then known to contract polio other than primates. Even though they had opened the door to further experimentation and the development of a vaccine against polio, Loring tempered the excitement, cautioning that the path ahead might still be long.

They were able to come up with a crude vaccine against polio in cotton rats later in 1947 before Schwerdt switched to the Virus Laboratory of the University of California at Berkeley. Here, he was able to further improve both his techniques and the product.

Working alongside his colleagues at Berkeley, Schwerdt developed a method to purify the poliovirus and also photographed it for the first time in pure form in 1953. He was involved in crystallising the pure virus in 1955 and also purified all three known major strains of poliovirus in 1957.“

https://www.google.com/amp/s/www.thehindu.com/children/loring-and-schwerdt-search-for-a-virus/article33410310.ece/amp/

This initial article gives credit to Loring and Schwerdt for the successful purification/isolation of Polio in 1946, with the caveat that it was taken from cotton rats and was only 80% pure. For further insight into this “historical” moment, here is Loring and Schwerdt’s original paper claiming this 80% purity:

Paper Isolation of a Macromolecular Constituent with Properties of the Lansing Strain of Poliomyelitis Virus

“It has been reported from this laboratory that highly active preparations of the MV strain of poliomyelitis virus could be obtained by differential centrifugation of clarified extracts of infected medullae-cords of rhesus monkeys. Application of the same procedures to brains and spinal cords of cotton rats infected with the Lansing strain resulted in appreciable quantities of high molecular materials which, however, showed only slight activity. Various modifications of the original procedure have been used in attempts to obtain highly active virus. It has been found that if the ether-clarified, aqueous extracts are frozen before they are subjected to ultracentrifugation, virus is obtained which is from 100 to 10,000 times as active on a nitrogen basis as the original clarified extract.

In the modified procedure the ether-clarified extract is prepared as previously described with the exception that filtration through celite is omitted. The slightly turbid solution is stored in 100 ml pyrex centrifuge bottles in a solid-C02 chest at from -35″ to -60°C for at least 5 days and then allowed to thaw at room temperature. Before the ice present has completely melted, the solution is centrifuged in the cold room at 4OC in an angle centrifuge, and the supernatant liquid is removed; the sediment is extracted in the centrifuge twice with M/l5 dipotassium hydrogen phosphate, and the latter extracts combined with the first supernatant liquid. The extracts obtained in this way are crystal clear, and depending on the length of time they have been frozen, contain from 75 to 51% of the nitrogen present before freezing. The appreciable amount of nitrogen precipitated is .protein in nature, and as the specific activity of the extracts after prolonged freezing is the same or greater than before freezing, it appears that normal protein rather than virus is denatured during this treatment. The results of a typical experiment showing the effect of freezing for various lengths of time on the yield of soluble nitrogen and on specific activity are shown in Table I. Activity is expressed as the 50% infective dose (I.D.50) in young cotton rats and is calculated by the method of Reed and Muench. The 50% infective dose is defined as the amount of virus nitrogen which produces definite signs of poliomyelitis within 2 weeks in 50% of cotton rats from 4 to 6 weeks old injected intracerebrally with 0.05 ml of inoculum in 0.10 hl acetate buffer at pH 4.“

The purified virus is obtained by subjecting the clear extracts to 3 or 4 cycles of ultracentrifugal purification as previously described. When tested for specific activity, samples from different experiments have given values for I.D.50 of from 10-^10.1 to 10-^8.3 g of virus nitrogen. The purified material, which is present after the third ultracentrifugation as an amber-colored gel, dissolves in water to form opaque, slightly yellow-colored solutions, which have failed to show any evidence of streaming birefringence at concentrations where this phenomenon is readily detected with tobacco mosaic virus. Similarly, the sedimented gel is completely isotropic when viewed between crossed Nicol prisms in contrast to similar gels of tobacco mosaic virus which, as is well known, are highly birefringent. Solutions containing 0.2 mg of virus nitrogen per ml give a faint Millon’s test for protein and a positive carbohydrate test with the orcinol-sulfuric acid reagent.

The yield for third cycle virus has been from 0.1 to 0.2 mg of virus nitrogen per 100 g of pooled brain and spinal cord. When the same purification procedures are applied to extracts of normal cotton rat brain and spinal cord, the percentage yields of macromolecular nitrogen after the extracts have been frozen for 5 and 10 days are of the order of 0.02 mg and 0.01 mg respectively. If the normal protein is present in the infectious extracts to the same extent it is found in normal tissue, it is evident that the maximum amount of normal macromolecular impurity present in the purified virus is of the order of 5 to 20%.”

Conclusions. The results outlined above including (a) the presence in the purified samples of a uniform and high specific activity (b) the demonstration that normal macro-molecular constituents are largely eliminated by the purification procedures, and (c) the fact that the samples are relatively monodisperse in the transparent ultracentrifuge provide strong evidence that the preparations consist of essentially pure Lansing virus.”

doi: 10.3181/00379727-62-15452.

As can be seen from this study, Loring admits that normal macromolecular impurity exists within his “pure” sample of the Lansing strain. He believes it is strong evidence that the preparations are “essentially pure” Lansing “virus.” However, there is no such thing as essentially pure. It is either pure or impure. This study can not be claimed as evidence of a purified/isolated Lansing Polio strain as any of the estimated macromolecular impurities could potentially be the “virus” particles they were searching for as the cause of disease.

This next paper was published not long after the previous one. In it, Loring admits that the images obtained of his “purified virus” are not uniform or consistent and range in size from 12 to 34 mp. Loring goes on to talk about the differences his team observed over other teams attempting purified/isolated Polio “virus.” It is clear from Loring’s comments that there are other asymmetrical or thread-like particles that Gard observed which were not in his own images. However, he admits there were small spherical particles in Gards images as well which line up with his work. Thus it was decided that these small spherical particles are the “Lansing virus” even though there are other particles admitted to be in the sample which could also be the “virus.”

Electron Microscopy of Purified Lansing Virus

“Eleven different samples of purified Lansing virus prepared as described in the preceding paper have been examined in the Stanford election microscope. The preparations were obtained from extracts that had been frozen from 4 to 26 days and gave specific activities, I.D. So, from 10-^10.l to 10-^8.3 g of virus nitrogen. Mounts were prepared by the usual technic of applying a small drop of dilute solution (10-^8 to 10^7 g of virus nitrogen per ml) to the collodion film and allowing the solution to dry or by washing away the remainder of the drop with distilled water after 2 to 5 minutes. The mount was then either introduced directly in the microscope or it was treated by the gold shadowing technic of Williams and Wyckoff before placing in the microscope.

Although all the micrographs have not yielded uniform or conclusive results, they have in general shown the presence of spherical or possibly slightly asymmetrical particles ranging in size from 12 to 34 mp.

Examples of micrographs obtained when the virus was examined without gold shadowing as well as of those obtained by the gold-shadowing technic are shown in Fig. 1a and 1b. Fig. 1a was obtained from an extract that had been frozen for 26 days and purified by 2 cycles of ultracentrifugal purification and Fig. 1b from an extract frozen for 4 days and purified by 3 cycles of ultracentrifugal, purification, the specific activities being I.D.50 = in both cases. Fig. 1a was obtained by the usual technic and Fig. 1b after gold shadowing.

In Fig. 1a there is some indication of slightly asymmetric particles with an average width of about 16 mp and an average length of about 31 mp or an average particle diameter of 23 mp. In Fig. 1b this is not so apparent and the range of particle size as measured by shadow width is from about 15 to 34 mp with an average value for particle diameter of 25 mp. The slightly lower value for the non-shadowed specimen can probably be explained by the relatively low contrast between particles and background with the accompanying lack of definition in the particle image. Under these conditions the measured dimension is the greatest distance within the particle image of points of appreciable photographic contrast, which may not coincide with the true edges of the particle. In the case of the gold-shadowed specimens the shadow width is the measured dimension and because of the high degree of contrast this can be determined with considerably greater accuracy.

Of particular interest is the absence in all of the purified Lansing virus preparation of asymmetrical or thread-like particles as found by Gardt for both murine (Theiler’s virus) and human poliomyelitis virus and as suggested by Bourdillon for the SK mouse virus. While the results of Gard appear conclusive in the case of Theiler’s virus, his conclusions with respect to human poliomyelitis were admittedly based on only a few experiments and open to other interpretations. The latter purified preparations from both infected tissues and feces contained components with sedimentation rates of the order of magnitude described in the preceding paper for the Lansing virus. Similarly while some filamentous particles were observed in his electronmicrographs, there was also ample evidence for small, approximately spherical, particles. We feel, therefore, that Gard’s results on human poliomyelitis are not necessarily incompatible with those presented here.

Conclusions. The results mentioned in the preceding paper as well as those given here lead to the conclusion that the Lansing virus, unlike Theiler’s virus and probably the SK mouse virus, is a relatively spherical or slightly asymmetrical particle of about 25 mp average particle diameter. It may be recalled that a somewhat similar conclusion was reported for the MV virus. It appears to the present authors that such values for the size of these strains of poliomyelitis virus of human origin are also more in accord with the filtration end-point data and the conditions that have been found necessary for sedimentation in the ultracentrifuge.”

doi: 10.3181/00379727-62-15453.

After the experiments with Loring, Carlton Schwerdt went off to attempt to achieve further purity of the “virus.” A few sources presented below discuss Schwerdt’s efforts. What is interesting to note here is that both sources admit to the lack of complete purification of the samples from the previous efforts by Loring, which only achieved purification of 1% of the “virus.” In discussing Schwerdt’s later work with researcher Howard Bachrach, it is claimed they were only able to achieve 10% purity. Only being able to purify 10% of a Polio “virus” is not purification at all. It seems we are once again stuck at “essentially pure,” however even that improbable statement is a stretch:

Howard L. Bachrach, 88, Early Polio Researcher, Is Dead

“With another biochemist, Carlton E. Schwerdt, at the University of California, Berkeley, Dr. Bachrach developed a method to isolate two strains of the virus and purify them so they could be studied under an electron microscope. Past efforts had been able to purify 1 percent of the virus; the Bachrach and Schwerdt research had achieved a purification level of 10 percent.

The greater purity enabled others to develop more exacting tests for the efficacy of polio vaccines. In 1955, Dr. Jonas Salk introduced a successful vaccine, known as the “killed virus” vaccine, using a form of the polio virus itself.”

“In 1954, before discovering the FMD vaccine, Bachrach left a lasting mark on the fight against polio. Working with Carlton Schwerdt, the two were able to isolate and “purify” 10% of the polio virus, cutting out the peripheral junk that prevented scientists from studying the disease. Researchers previously had only been successful in purifying 1% of the virus, and the additional clarity led to several more advancements, culminating in 1952 with Jonas Salk’s vaccine.”

https://www.nationalscienceandtechnologymedalsfoundation.org/laureates/howard-l-bachrach

Notice in the above article that the word “PURIFY” is in quotations. Even after several years, Schwerdt was only able to achieve a lackluster 10% purification of the Polio “virus.” Sadly, that did not stop the misguided headlines claiming purification/isolation of a Polio “virus.” Here is the summary from his 1953 paper:

Purification Studies on Lansing Poliomyelitis Virus

“Purified concentrates of the Lansing strain of poliomyelitis virus have been prepared from cotton rat CNS with specific infectivities increased 20,000 times, on the average, above that of the original infected tissue. The fraction of total virus infectivity recovered in the purified concentrates averaged 40 to 50%.

The virus in the concentrates of high specific infectivity has been identified by analytical electron microscopy to be a spherical particle 28 mμ in diameter. This identification was made through correlative experiments relating particle counts to infectivity and is discussed in detail. The concentration of the virus in CNS tissue of paralytic cotton rats is calculated to be about 0.2 μg per gram of tissue. It is shown that about 21,000 virus particles or 3.0 × 10^-13 g of virus are consistently present in one LD⁵⁰ inoculum for cotton rats, but it is not known whether all or a constant fraction of these particles are infectious. The electron microscopic evidence shows that the virus is highly uniform in size and shape.”

https://www.jimmunol.org/content/72/1/30.long

Unfortunately, I was unable to access the full article as it is blocked behind a paywall. However, highlights from a study by Schwerdt a year later gives a good idea of what was done.

From this study, you will see that the images and the assumed “virus” come from monkey kidney tissue cultures which have been passaged several times. The sample they are working with does not come from a human and it already starts off as an unpurified mixture. It is clear that Schwerdt guesses as to a certain size/shape for the particles in the EM images and assumes that they are the “virus” he was searching for without direct evidence to this being the case. However, even with his assumption, he admits that they are uncertain that these paticles are in fact the Polio “virus.” He estimates size/shape based on various measures but there are several limitations he notes such as the presence of artefacts, possible technological limitations, and various assumptions which were made. Schwerdt also states that the images obtained are in direct contrast to images of Polio taken by Sabin. In other words, there is no proof within this paper that these images are of a “virus” at all, let alone a “purified” one:

Morphology of Type I1 Poliomyelitis Virus (MEFl) as Determined by Electron Microscopy

“Previous attempts to determine by electron microscopy the size and shape of particles of human poliomyelitis virus have been equivocal because of uncertainty as to the identity of the virus particle among various objects of similar appearance found in the electron micrographs (1-6). Improved methods of purification applicable to poliomyelitis virus propagated in central nervous system tissue in vivo, or in monkey kidney tissue culture, now yield virus concentrates containing at most 2 classes of particles and frequently only one( 7,8). Furthermore, the examination of such purified virus concentrates by analytical electron microscopy has resulted in the establishment of a constant ratio of numbers of the particle believed to be the virus to the infectivity titer (8). In a preliminary determination this particle has been found to be approximately spherical and about 28 mp in diameter, but its successful identification seems to justify at this time a more definitive examination of its shape and size.

Materials and methods. The virus used in this study is a tissue culture MEFl strain received from Dr. Jonas Salk in 1952. It has been passed 3 times in monkey kidney tissue culture in this laboratory, and the third passage has been used as a virus seed pool for the production of liter quantities of tissue culture virus suspension for purification purposes. The method of purification employed was similar to that described by Bachrach and Schwerdt for the purification of Lansing virus from infected cotton rat brains and cords (8).”

Fig. 2 illustrates a frozen-dried specimen of purified virus. The very small, irregularly shaped particles seen over the background are believed to be non-volatile impurities in the ammonium bicarbonate solution, since similar debris is observed in ammonium bicarbonate blanks.”

Discussion. Individual particles of XEFl poliomyelitis virus appear to be spherical in frozen-dried preparations and to have the shape of slightly flattened spheres in air-dried mounts. The high uniformity of particle diameter is demonstrated in Fig. 1 where rows formed by packing are seen to be straight over distances including a dozen or so particles. Diameter measurements of individual frozen-dried particles offer further evidence of uniformity in that the variation of the measured diameters is not unequivocally greater than the estimated uncertainty of measurement. These conclusions are in distinct contrast to the results published by Sabin, Hennessen and Warren(6) for a type I1 virus where the electron micrograph shows a great variation of particle size.

The absolute value of the diameter of the virus particles is subject to several uncertainties, owing to possible artifacts of preparation, the finite resolving power of the electron microscope, the calibration of magnification, and the limitations of the measurement of the photographic images. If we assume that no appreciable shrinkage of individual particles has occurred during the formation of an array like Fig. 1, the average diameter of the particles measured in a row is least susceptible to errors of image formation and measurement.

If individual particles are measured, the most reliable diameter is probably secured from the frozen-dried preparations where no sign of surface-tension distortion is seen. Our estimate of the uncertainty in the value given for the particle diameter is +-2 mp.

Summary. The particles of the MEFl poliomyelitis virus, grown in monkey-kidney tissue culture, and subsequently purified by chemical and physical means, are spherical in shape and highly uniform in diameter. The diameter of individual particles, prepared by normal air-drying from a 0.1 31 ammonium acetate suspension, is found to be 31 mp. The diameter of particles packed in a crystalline array, or frozen-dried from a 0.1 31 ammonium bicarbonate suspension, is found to be 27 mp. It is believed that the latter diameter is the more significant and reliable figure.

doi: 10.3181/00379727-86-21082.

In Summary:

• Hubert Scott Loring and Carlton Everett Schwerdt announced their “successful isolation” of poliovirus on January 10, 1947
• However, it was only 80% pure and extracted from cotton rats infected with polio
• Loring was heavily  involved in two major areas during this time – one being the purification of the poliomyelitis “virus” (which is an admittance that all the work from 1908 to 1947 proceeding his supposed purification was with unpurified material)
• Along with his student Schwerdt, Loring spent three years searching for the poliovirus
• Loring and Schwerdt were able to obtain the “virus” with 80% purity
• They did not “isolate” Polio from humans but from cotton rats
• Loring tempered the excitement, cautioning that the path ahead might still be long
• Schwerdt developed a method to purify the poliovirus and also photographed it for the first time in pure form in 1953 (the particles assumed to be Polio were never photographed in “pure” form for 45 years)
• He was involved in crystallising the “pure virus” in 1955 and also “purified” all three known major strains of poliovirus in 1957 (up until 1957, all 3 Polio strains had never been “purified”)
• It was later revealed that the efforts by Loring had been able to purify only 1 percent of the “virus”
• The Bachrach and Schwerdt research, which set out to achieve greater purity than Loring, only achieved a purification level of 10 percent

• Application of differential centrifugation to brains and spinal cords of cotton rats infected with the Lansing strain resulted in appreciable quantities of high molecular materials which, however, showed only slight activity
• Various modifications of the original procedure were used in attempts to obtain highly active “virus”
• It was found that if the ether-clarified, aqueous extracts are frozen before they are subjected to ultracentrifugation, “virus” is obtained which is from 100 to 10,000 times as active on a nitrogen basis as the original clarified extract
• Loring’s experiments essentially consisted of freezing, thawing, and centrifuging the pooled brain/spine tissue culture supernatant and measuring nitrogen levels to define “purity”
• In his modified procedure, the ether-clarified extract is prepared as he previously described with the exception that filtration through celite is omitted
• He further described the process in this way:
  • The slightly turbid solution is stored in 100 ml pyrex centrifuge bottles in a solid-C02 chest at from -35″ to -60°C for at least 5 days and then allowed to thaw at room temperature
  • Before the ice present has completely melted, the solution is centrifuged in the cold room at 4OC in an angle centrifuge, and the supernatant liquid is removed
  • The sediment is extracted in the centrifuge twice with M/l5 dipotassium hydrogen phosphate
  • The latter extracts are combined with the first supernatant liquid
• The results of a typical experiment showed the effect of freezing for various lengths of time on the yield of soluble nitrogen
• “Virus” activity was expressed using the same end dilution guesstimate Enders used later in 1949
• The 50% infective dose (I.D.50) in young cotton rats and was calculated by the method of Reed and Muench
• The 50% infective dose is defined as the amount of “virus” nitrogen which produces definite signs of poliomyelitis within 2 weeks in 50% of cotton rats from 4 to 6 weeks old injected intracerebrally with 0.05 ml of inoculum in 0.10 hl acetate buffer at pH 4
• The yield for third cycle “virus” was from 0.1 to 0.2 mg of “virus” nitrogen per 100 g of pooled brain and spinal cord
• When the same purification procedures are applied to extracts of normal cotton rat brain and spinal cord, the percentage yields are of the order of 0.02 mg and 0.01 mg respectively
• Due to the nitrogen yields, Loring guesses that the maximum amount of normal macromolecular impurity present in the “purified virus” is of the order of 5 to 20% (which would indicate that the sample is not completely purified/isolated)
• He states that he demonstrated that normal macro-molecular constituents are largely eliminated (i.e.not entirely eliminated) by the purification procedures
• Loring concludes that he provided strong evidence that the preparations consist of essentially pure (i.e. not pure) Lansing “virus”

• Eleven different samples of “purified Lansing virus” prepared as described in the preceding paper were examined in the Stanford election (not a typo) microscope
• All of the micrographs did not yield uniform or conclusive results
• They showed in general the presence of spherical or possibly slightly asymmetrical particles ranging in size from 12 to 34 mp
• Two images are submitted (obviously being considered the best representation they could achieve): Fig. 1a was obtained by the usual technic and Fig. 1b after gold shadowing
• In Fig. 1a there was some indication of slightly asymmetric particles (having parts that fail to correspond to one another in shape, size, or arrangement; lacking symmetry) with an average width of about 16 mp and an average length of about 31 mp or an average particle diameter of 23 mp
• In Fig. 1b this was not so apparent and the range of particle size as measured by shadow width is from about 15 to 34 mp with an average value for particle diameter of 25 mp
• Loring noted the absence in his samples of asymmetrical or thread-like particles as found by Gardt for both murine and human poliomyelitis “virus” and as suggested by Bourdillon for the SK mouse “virus”
• Gardt’s conclusions with respect to human poliomyelitis were based on only a few experiments and open to other interpretations
• Loring states that while some filamentous particles were observed in Gardt’s electronmicrographs, there was also ample evidence for small, approximately spherical, particles
• Thus, since Loring did not have filamentous particles yet he and Gardt both had spherical particles, he feels their results are compatible
• Based on his own findings, Loring concludes Polio is a relatively spherical or slightly asymmetrical particle of about 25 mp average particle diameter

• Previous attempts to determine the size and shape of particles of Polio by EM have been equivocal (open to more than one interpretation; ambiguous, uncertain or questionable in nature) because of uncertainty as to the identity of the “virus” particle among various objects of similar appearance found in the electron micrographs
• Improved methods of purification applicable to poliomyelitis “virus” propagated in central nervous system tissue in vivo, or in monkey kidney tissue culture, now yield “virus” concentrates containing at most 2 classes of particles and frequently only one
• The “virus” used in this study is a tissue culture MEFl strain received from Dr. Jonas Salk in 1952
• It had been passed 3 times in monkey kidney tissue culture in their laboratory, and the third passage has been used as a “virus” seed pool for the production of liter quantities of tissue culture “virus” suspension for purification purposes
• In the second of two EM images provided, it is stated the very small, irregularly shaped particles seen over the background were believed to be non-volatile impurities in the ammonium bicarbonate solution
• Diameter measurements of individual frozen-dried particles offer further evidence of uniformity in that the variation of the measured diameters is not unequivocally greater than the estimated uncertainty of measurement
• Their conclusions are in distinct contrast to the results published by Sabin, Hennessen and Warren for a type I1 “virus” where the electron micrograph shows a great variation of particle size
• The absolute value of the diameter of the “virus” particles is subject to several uncertainties:
  1. Possible artifacts of preparation
  2. The finite resolving power of the electron microscope
  3. The calibration of magnification
  4. The limitations of the measurement of the photographic images
• Their estimate of the uncertainty in the value given for the particle diameter is +-2 mp.
• Based on two different methods, the particles ranged in size from 27 to 31 mp and they believed that the latter diameter is the more significant and reliable figure

When the headlines claim purification/isolation, it is always found out after closer inspection that this is never the case. Nowhere in the work of Loring and Schwerdt did particles claimed to be the Polio “virus” ever come from humans. The supernatant came from tissue cultured brains/spines of cotton rats or monkey kidneys. There were admittedly other asymmetrical particles in the EM images which is the exact opposite of isolation. It’s apparent just from looking at the work of both Hubert Loring and Carlton Scwerdt that the complete purification/isolation of a Polio “virus” never took place. It is admitted that at most, only 10% purity was ever achieved. This “purification” was said to set up the creation of the vaccine yet it is clear that the vaccine came from impure particles assumed to be “virus” taken from monkey kidney cultures. This in effect caused many health issues due to the admitted impurity of the monkey SV40 “virus” (which is just foreign animal DNA) contained within the tissue cultured supernatant used for the Polio vaccine production from 1955 to 1961.

Virology loves to proclaim purification/isolation of “viruses” yet the problem is they never actually do either of these things. The beauty of Polio is that we have the ability of hindsight and we can not only look at the researchers work but also the admittances made later. The headlines in 1947 proudly stated that “Farm Scientists Isolate Polio Virus.” Loring’s second paper was titled “Electron Microscopy of Purified Lansing Virus.” Schwerdt published a paper with the heading “Purification Studies on Lansing Poliomyelitis Virus” with the first sentence claiming purified “virus” prepared from cotton rat CNS. However, as stated previously, this “purification” ranged from a measly 1 to 10% of the alleged “virus.”

“Essentially pure.”

Or in other words, not pure at all.

Maybe this embarrassing proclamation as to the purification/isolation of Polio which later turned out not to be is the reason why Loring and Schwerdt have been stricken from the Polio discovery process? The rest of the “researchers” should be as well.