BSK culture experiment

different size spiral agents


 

Large numbers of these tiny spirals have been observed. If some of the agents are spirochetes, the question arises - where did they come from - if it is accepted that failure to locate these agents in a fresh blood sample indicates they were not originally present?

A possible answer is that they are spirochetes growing from round-bodies (endospores/cysts) and that they grow as very tiny spirals that gradually elongate and thicken, increasing thier wavelength and pitch as they do so. I should note here that I have observed spiral agents appearing to emerge from round-bodies that had dimensions matching the larger subject in the first picture.

However; a professor of microbiology remarked that bundles of helical (external) flagella can clump together and appear like a spirochete. The cultures I use are generally over 5 months old, so it seems possible that lots of bacteria have grown and died - leaving behind thier flagella. But this raises some further questions:

If these are bundles of flagella, then why, even when they are attached to a round-body do they sometimes appear to thicken away from the body?

Why do they invariably appear to have an even thickness or appear thicker in the central region and taper, often quite symetrically, with no apparent raggedness towards each end?

The tiny spirals are often attached to a spheroidal or elongated body at one or both ends. Infrequently, I have observed what look like 2 spirals roughly bound together. Could bundles of helical flagella look like this?

Duplicated from my previous BSK Experiment page:

Photo with measurements and comparisson to borrelia burgdorferi dimensions; labels refer to the scale dimension of the lines:-

Image showing different dimensions of 2 spiral forms. Although I previously observed that when a slide dries out the spirals sometimes appear stretched; it does not look as though this would account for the discrepency in sizes of these 2 subjects (dimensions in microns):-

The following shows a wet-slide stained with acridine orange. The micrographs are edited to match but the dark-blue filter actually required a much longer exposure time.

In this micrograph there are 18 tiny spiral agents and one 'normal' sized (as per b. burgdorferi spec). The tiny spirals were not observed elsewhere on the slide. This raises the question of whether they are associated with the large spiral. Did they grow from it; or did it's presence trigger growth from cysts by quorum sensing; or were they attracted to it? None of the spirals exhibited motility.

The following 2 images show tubular-looking structures containing coccoid or slightly elongated bodies. The spirals are firmly attached to or emerging from the long 'tube'. I generally ignore tubular structures because of the problem of differentiating them from filamenting bacteria (not 'filamentous'). As filamenting is a manifestation of many types of bacteria under adverse conditions, yet can still support reproduction within the elongated tube of the cell-wall, it suggests that it is an option that might occur in adverse, but not terribly adverse conditions; i.e. adequate nutrients must still be available for growth. Borrelia and other spirochetes are known to be able to encyst in adverse conditions, but this requires the presense of flagella and functional motor. Perhaps, when lacking these, the 'pre-filamented' (so-to-speak) elongated body of spirochetes can perform the same trick as filamenting bacteria by reproducing within the cell wall and generating a string of coccoids destined to become 'baby' spirochetes.

 

I frequently observe the presence of what look like short chains of coccoid or slightly elongated coccoid bodies and tend to ignore them as being unrelated to spirochetes or such that I cannot demonstrate a connection. The 2 following micrographs show 2 long bodies that appear quite different in structure. They are firmly connected at the juncture which was also firmly attached to the slide surface (thorough 'prodding' with the objective lens proved this).

Micrograph below taken at 18:44 - i.e. 2 hours after the photo above. The spirallad-looking agent appears to be separating and what appeared to be a chain of coccoids now appears to be 2 spirals

The juncture of these 2 spirals appeared like a droplet though it could be a sharp bend

Here are some micrographs of the very tiny spirochetes. Similarly sized spiros have been found growing from and attached to borrelia (see Lymebug videos on Youtube). However, these appear to be reproducing 'rhizomatously' which suggests they are mature. The chain of 3 below (was actually a chain of 4 with one out of focus) could be shown to be multiple spirochetes by judicious prodding with the lens which broke the chain into separate individuals. The dimensions of these spirochetes corresponds with treponema.

Last update July 26th 2011

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