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Working together back in 1931 and using one of the smaller Rife microscopes having a magnification and resolution of 17,000 diameters, Dr. Rife and Dr. Arthur Isaac Kendall, of the department of bacteriology of Northwestern University Medical School, were able to observe and demonstrate the presence of the filter-passing forms of Bacillus typhosus. An agar slant culture of the Rawlings strain of Bacillus typhosus was first prepared by Dr. Kendall and inoculated into 6cc. of "Kendall" K Medium, a medium rich in protein but poor in peptone and consisting of 100 mg. of dried hog intestine and 6 cc. of tyrode solution (containing neither glucose nor glycerine) which mixture is shaken well so as to moisten the dried intestine powder and then sterilized in the autoclave, 15 pounds for 15 minutes, alterations of the medium being frequently necessary depending upon the requirements for different organisms. Now, after a period of 18 hours in this K Medium, the culture was passed through a Berkefeld "N" filter, a drop of the filtrate being added to another 6 cc. of K Medium and incubated at 37 degrees C. Forty-eight hours later this same process was repeated, the "N" filter again being used, after which it was noted that the culture no longer responded to peptone medium, growing now only in the protein medium. When again, within 24 hours, the culture was passed through a filter - the finest Berkefeld "W" filter, a drop of the filtrate was once more added to 6 cc. of K Medium and incubated at 37 degrees, a period of 3 days elapsing before the culture was transferred to K Medium and yet another 3 days before a new culture was prepared. Then, viewed under an ordinary microscope, these cultures were observed to be turbid and to reveal no bacilli whatsoever. When viewed by means of dark-field illumination and oil-immersion lens, however, the presence of small, actively motile granules was established, although nothing at all of their individual structure could be ascertained. Another period of 4 days was allowed to elapse before these cultures were transferred to K Medium and incubated at 37 degrees C. for 24 hours when they were then examined under the Rife microscope where, as was mentioned earlier, the filterable typhoid bacilli, emitting a blue spectrum, caused the plane of polarization to be deviated plus 4.8 degrees. Then when the opposite angle of refraction was obtained by means of adjusting the polarizing prisms to minus 4.8 degrees and the cultures illuminated by a monochromatic beam coordinated in frequency with the chemical constituents of the typhoid bacillus, small, oval, actively motile, bright turquoise-blue bodies were observed at a magnification of 5,000 diameters, in high contrast to the colorless and motionless debris of the medium. These observations were repeated eight times, the complete absence of these bodies in inoculated control K Media also being noted.
To further confirm their findings, Drs. Rife and Kendall next examined 18-hour-old cultures which had been inoculated into K Medium and incubated at 37 degrees C., since it is just at this stage of growth in this medium and at this temperature that the cultures became filterable. And, just as had been anticipated, ordinary dark-field examination revealed unchanged, long, actively motile bacilli; bacilli having granules within their substance; and free swimming, actively motile granules; while under the Rife microscope were demonstrated the same long, unchanged, almost colorless bacilli; bacilli, practically colorless, inside and at one end of which was a turquoise granule resembling the filterable forms of the typhoid bacillus; and free swimming, small, oval, actively motile, turquoise-blue granules. By transplanting the cultures of the filter-passing organisms or virus in to a broth, they were seen to change over again into their original rod-like forms.
To further confirm their findings, Drs. Rife and Kendall next examined 18-hour-old cultures which had been inoculated into K Medium and incubated at 37 degrees C., since it is just at this stage of growth in this medium and at this temperature that the cultures became filterable. And, just as had been anticipated, ordinary dark-field examination revealed unchanged, long, actively motile bacilli; bacilli having granules within their substance; and free swimming, actively motile granules; while under the Rife microscope were demonstrated the same long, unchanged, almost colorless bacilli; bacilli, practically colorless, inside and at one end of which was a turquoise granule resembling the filterable forms of the typhoid bacillus; and free swimming, small, oval, actively motile, turquoise-blue granules. By transplanting the cultures of the filter-passing organisms or virus in to a broth, they were seen to change over again into their original rod-like forms.
