Variety in Microbial Morphology

One of the clear features of modern microbiology and microbiologists is that they are members of what we call the ‘monomorphist school’. They believe that bacteria are very simple organisms. What I want to show in the next half an hour or so is that, in fact, bacteria are very complex morphological organisms and they do vary quite considerably. I am going to mention Borrelia, of course, but I am also going to mention other diseases and even cancer later on in the talk.

If you look back at the history of microbiology; by the 1880s microbiology was beginning to settle down as a subject. Micro-organisms had been discovered in the 1820s / 1810 by people by Erinberg and they had done a lot of work on the morphology and the classification but it was really people like Koch and Ferdinand Cohn who really established bacteriology as a systematic subject. And what they did, was that they were interested in finding out the cause of disease. If you had tuberculosis, you wanted to find an organism, you wanted to show that organism caused TB and you wanted to find a cure, if possible, or some kind of immunisation against the disease, against the bacteria, and this is, of course, the basis of what we call Henley - Koch's postulate. The idea that one organism causes one disease. This lead to the idea of diagnostic microbiology. All you have to do is find one organism and you find the cause of disease and ultimately cure it.

Now, basically, this idea of monomorphism is based on a very simple thing, that bacteria are simple. Here we have a classic picture of bacillus. You can see it is a rod shaped organism, there are no fancy structures to it, there is nothing complicated about it, there may be a flagellum, there may be little hairs coming off which we call fimbriae, but generally it is a very straightforward, single monomorphic organism and this idea has spread throughout biology, throughout modern bacteriology today. So, if you look at the textbooks, the kind of thing that we teach our students in the first year course in bacteriology, microbiology, you find that we talk about simple bacteria: cocci, fibriole, bacilli, spiral, spirochaete,. We teach our students that these are simple organisms. They stay like that, they don’t change, they never become anything else other than a coccus, a coccus remains a coccus, a bacillus remains a bacillus. There is nothing complicated about it and all you have to do is to look for these organisms and you draw them and you pass your exams and everybody is happy. Often, of course, you are drawing an air bubble until the demonstrator tells you that, it’s not an air bubble you are looking for, it’s a coccus. But this is basically what we are teaching our students.

So, again, we may see a bacterium doing simple division. There is nothing really complicated about it, we can do a DNA stain. And we can do now a lot of molecular techniques, a lot of fancy microbiology, fancy biochemistry on these organisms here we can really go to town on their DNA and so on. In fact, very few microbiologists actually look at organisms under the microscope. Increasingly, we are reducing the time we spend with our students using the microscope. For example, we used to spend a whole day with our first years looking at the taxonimy of the fungi, we go through the whole group: Phyomycetes, Ascomycetes and so on. Now we don’t bother with that because there is too little time for that, we are running gels, we are going to be doing 16S whatever. So we don’t have time to do these basic bacteriological, microbiological studies.

Occasionally, a bacterium will throw up a spore. A spore is a survival structure and this will then give rise to a new bacillus and this is all very simple, a very nice idea of bacteriology. This is what we call ‘ monomorphism’. Now, very early in microbiology, and kind of going on in the background for the last 100 years, there are what we call ‘pleomorphists’; the idea that bacteria are far more complicated than the simple view point that we have got here. Many people in the past have suggested that bacteria, even the common bacteria, have life cycles. They don’t stay as they are, they change: a coccus may change into a rod, a rod may go to a coccus, they may become filamentous, they may get quite extravagant in their morphologies. Now this has not been accepted by most of the mainstream because, as I said, the mainstream are educated and brought up with this monomorphic viewpoint.

Now there is an extreme view of pleomorphism which kind of soils the water a bit for us standard microbiologists as it were who are interested in pleomorphism. And this is by a German scientist called Enderlein. Now Enderlein is one of the proponents of this idea of cyclogony - extreme, extreme, extreme, pleomorphism. And unfortunately, most microbiologists, including myself, who are amenable to this, can’t really get into this story because of all the complex terminology. It’s almost like reading Lord of the Rings when you read this kind of stuff. It does not feel scientific because of all the strange terminology involved. But there are proponents, disciples of Enderlein, and I am not suggesting that it is wrong for a minute, I am just suggesting that for most modern microbiologists it’s very difficult to understand.

Now, there are some life-cycles in the literature. Here is one for a bacterium called Spiroplasma mirum and you can see that this has been published in the standard literature and we can see here that this bacterium is supposed to go through some very complex stages. It starts off as a little rod, and then it goes more complex. These are very much like the things that Andy Wright and others have seen down the microscope, I guess, in Borrelia or in ME patients, and we get this complexity here. Now, clearly, this is not the kind of thing that you generally get in textbooks. You would not find this life-cycle in a basic microbiology textbook saying that bacteria go through life-cycles because this is just not what is acceptable although it has been accepted and published in the mainstream literature.

Similarly, here we have an example, and you have seen a lot of these, of so called spirochete life-cycles. When spirochetes were first discovered, they were regarded as protozoans and they are supposed to have a life-cycle developing from little blobs through to these spirochete forms and then they develop different forms when they get inside the cell and so on. And this is very much analgious to what we think about in Borrelia. This has been completely neglected, all of this work from the early 1900s, done by a very good microbiologist, very good bacteriologist, using a light microscope, admittedly, spending a lot of time and these guys were very good at microscopy so we should really take their word more seriously than we do because at least they could use a microscope, at least they could focus a microscope, which most microbiologists these days can’t do. So that really is the spirochete. This is the form that everyone works with, this is what we can understand, we can see, and we don’t really need to worry about the life-cycle. If we can find the spirochete, we can diagnose the disease.

There are other life-cycles which are accepted in the literature. Chlamydia, for example, goes through a complex life-cycle. Rickettsia is another one but these are kind of regarded as ‘strange diseases’, unusual, these are bacteria that do these things and they are put to the back of the text book because they are not really simple monomorphic organisms, they are best left alone, best ignored because they are not simple and this is really the view taken by many microbiologists, I am afraid to say.

It becomes very apparent when you start working with micro-organisms that they do change shape. This is E.coli, the famous rabbit of the bacteriological world, the most boring organism, supposedly, on the planet in terms of morphology. All that you are supposed to get normally is a rod - shaped structure like this. This is what you get if you grow this bacterium in high nutrient media: nutrient broth - LB medium, but look what happens when you deny it nutrients: it starts to filament. This is the same organism, this is not a different organism, some of these bacteria are beginning to filament. So this is about 40 microns in length, same bacteria, and what happens is, in fact, we know the genetics of this gene involved, is that this bacterium divides in the normal way but instead of separating, the bacteria continue to divide and form this filament. Now this filament can happen when you starve it, it can happen when you add antibiotics (penicilin does this to E.coli, for example), metals will do this. In fact, if you weren’t taught that E.coli was a rod -shaped organism and you actually worked with the organism you might think that it was, in fact, a filament that occasionally forms these rods. It depends on your perspective, where you are coming from. Quite clearly, this organism here, or this structure, is the same organism, genetically it is the same organism in terms of that it might have different cells or proteins perhaps but the 16S will be the same, the same organism as this. Clearly, if this gets inside your body, if this gets inside your gut, or this is forming a bio-film on your teeth, or anything, this will act completely different to this because it is entrapped by other organisms and it will form a mesh and so on and form a complex bio-film. This structure doesn’t occur in textbooks because this is E.coli as we know it. When you see these, it is really exciting.

Here is another example, often we just see them as smaller versions of this but here we’ve got one which is actually forming a ring which is very interesting. Now these are under scanning EM, scanning electron microscope pictures, and this explains a lot of problems that arose with light microscopy because if you look at this under the light microscope you might think you had got a structure that was filamentous with a kind of ring forming on it: you might think this was an unusual formation and you might call it something really exotic and think it was part of the life-cycle but as we see under the electron microscope it is really the filament bent over itself to form a ring like that. So the electron microscope can certainly sort out what the problem is. There were a lot of papers in the early literature, for example, that say that E.coli will form T-shaped structures and basically, all they are when you see them, are one rod abutting off the side of another one. Its not actually T-shaped, it’s just two rods together. So clearly, there were mistakes in the early literature, we wouldn’t expect anything else.

As a kind of standard microbiologist, I then went to look at the Borrelia information. Went on to Google and just looked under Borrelia pleomorphism and this is what I got. These ideas of these cysts and various anomalies but really there is very little, at least on the Internet, about the complexities that we see under the microscope that Andy Wright is going to be talking about later. We have really kind of sanitised even this into one or two forms which we can say are survival structures like spores. We know that Borrelia has a long generation time and we know that it can act as a persistor, it can hang around in the body for a very long time. It persists even with antibiotic treatment and so on. So this is what I get from even the standard Google search, obviously, if you go into other sites, which we will talk about today, you will get more complex views of what Borrelia is. So this is what a standard microbiologist might get a view of Borrelia.

Staying with Leptospira, you get these granules forming along its length and, again back to Borrelia, these granules again which are supposed to be these survival structures. Nothing about the complex, amazing, structures that you see when you look at live blood. Because of course very few standard microbiologists look at live blood. We either isolate the organisms on to media or we do DNA work on them but very little blood work is done. This, of course, is where amateurs, as it were, if you want to call them that, or people in the medical profession, GPs like Andy, can take a tremendous role just by looking at these things which is something that a more standard microbiologist wouldn’t do.

And again, we have this viewpoint, that coccoid bodies give rise to spirochaetes but, again, who would need to worry about this when you can just find the spirochaete and diagnose it from the spirochaete. So the consensus that you get from the literature, if you don’t go deep into it, is that Borrelia is just a fairly ordinary organism, a fairly interesting little thing, it produces spirochetaes, it’s got these cysts, blobs, spheres and fisticules but it doesn’t really have a complex developmental cycle and all these things are just there to get it through some unfavourable conditions.

Now I have been working for the last ten years on the possibility that pleomorphic bacteria can cause cancer. If you think the Borrelia story is complex and controversial, try and get in to the idea that bacteria cause cancer. There is, in fact, a tremendous literature on this. It goes way back to the 1880s, the so- called cancer germ, the idea that bacteria cause cancer. And if we go to the most famous paper on this in the 1920s, we find, in fact, that Glover, isolated a bacterium, grew it up, which went through a life-cycle. It produced all kinds of weird and wonderful shapes, bacilli formed spores and then ballooned out, it produced these mycelial-like things, it produced these very fine filaments and all the way back from the coccoid. And at some point, it formed an invisible phase and also a filterable phase, it could go through a filter. I will talk about this in more detail in a minute. Now, whether this is a real cycle in the sense that each of these things must happen, I have my doubts, but it may well be that they can happen across...it doesn’t necessarily go from there to there, it may go from there to there and so on an so forth. So perhaps a cycle is a misnomer but this is how he has drawn it.

Of course, this has been generally regarded as contamination. Why wouldn’t it be? This person, Glover, who was an extremely good microbiologist, apparently was working with a tube full of about 20 organisms, presumably left the top off the tube for about 24 days and not realised that there were things called contaminants. Because to get that kind of contamination is almost impossible anyway, so that really ruins the idea of contamination. So if you could do some DNA work on that you might hopefully find that these structures were the same bacterium.

People have taken these cancer germs and claimed to have cured cancer in animals and in humans even with vaccines from cancer germs but again the cancer establishment does not accept in any shape or form, or minor shape or form, that bacteria cause cancer.

I wrote this paper. Somebody was talking earlier about getting into the standard literature and it is very difficult. This is an historical paper. I got all the literature together talking about highly pleomorphic staphylococci. Now staphylococci aren’t supposed to be pleomorphic, they are coccoid shaped organisms that don’t change much. There is a lot of literature saying that these highly pleomorphic Staph are found in cancer patients and tumours and I couldn’t get this published in the establishment literature so I went to a journal called Medical Hypotheses which is a superb journal for people like myself. They will take weird, weird, papers on the basis that these are hypotheses. I really recommend anybody who is interested in getting stuff published to go there. It is a highly regarded journal but it is very open to unusual thoughts.

Now, here is a most amazing thing. Helicobacter pylori, the organism which causes stomach ulcers and we know that stomach ulcers before 1984 were caused by ‘stress’ before Marshall and Warren came along and found that this bacterium in the gut causes stomach ulcers. This bacterium is also linked to stomach cancers. So if you talk about bacteria causing cancer, this is the only organism the cancer experts will mildly accept as a possibility: H pylori. So if you look under ‘cancer bacteria’ you will get very little, only my own papers, but if you go ‘H. pylori cancer’, you will get enough information to keep you going for years. So for some reason H. pylori has suddenly stopped being a bacterium and has become H. pylori but it is a bacteria and it is a very common bacteria, it is a very easy, simple bacteria, there is nothing particularly exciting about it except that it does form pleomorphic forms. There are these coccoid forms, this is normally a spiral organism but it forms coccoid forms. Now how does the establishment get over the fact that this organism which is very important is pleomorphic? Well, they say that the coccoid form forms later on, and is degenerate. This is an old idea about pleomorphism that anything which moves away from the norm is degenerate, what we call an ‘involution form’, it’s not real, it’s just something that happens. Even though you can keep producing it over and over again, it’s involution form. And many people say the coccoid form is dead, it’s just a dead organism: ‘This parrot is dead’, but in reality this parrot is by no means dead, it is alive and it will be very important, probably, in causing inflammation infecting the gut possibly forming stomach cancer.

So, here we have another bacterium which we think is linked with cancer which is pleomorphic. You can’t read all of this kind of stuff without wanting to get involved in it so I have been trying to look at tumours in humans and animals for the last ten years and I have been spectacularly unsuccessful not in terms of microbiology but in terms of getting tumours. One of the problems is getting supplies of human tumours. You would think that it would be relatively simple. Go to a doctor, go to a hospital, ask for a few bottles of tumours and you can cut them in half and play around until you are blue in the face. And this is what I though might happen when I was a naïve, younger man. In fact, of course, it is extremely difficult. There are these ethics committees and so on. And I have actually been on ethics committees and said and talked to people when they say “why do you want to do this work, Dr Wainwright?” (they are usually very condescending) and I say I am interested in the idea that bacteria may cause cancer. They say, well, we are not going to let you do this because bacteria don’t cause cancer. Why would we waste tumours on you when we know that bacteria don’t cause cancer? Why waste tumours on you? In fact, these tumours have more civil rights than the people that are killed and I am not joking – it gets to the point where if you squash a tumour, they might send the police around.

I even get people writing to me, women usually with breast tumours, saying I will donate my tumours but no, it is not allowed, they have got to be looked after. But anyway, fortunately, I had a Vet who for a few weeks and months, would supply me with dog tumours and then he realised that perhaps it may cause problems with his career. Supplying me with just dog tumours is a problem, apparently, I don’t know why, but that is what he thought. But while he gave me some dog tumours, I worked on them.

This is what we found in the mammary tumour of a dog and, I won’t go into the isolation procedure which is quite complex, here we get an organism which again shows pleomorphism. This is on medium, this is a rod shaped, filamentous, here is rod shaped forming filaments, here it is forming coccoid bacilli, more coccoid shapes but here is the interesting one. This thing here, if you were looking down the microscope for the first time at this, you might think this was a fungus. It looks filamentous, it’s got these filaments which appear to produce spores or something like that .

I was a micologist, a fungus expert, for 30 years, and if I was just looking at that blind, as it were, I would think it was a fungus. And this may explain why many people in the past have associated pleomorphic bacteria with fungi either as contaminants or they have even said that fungi change into bacteria, which is not the case, but you can see where they are coming from, where they are making a mistake. This is, in fact, a beautiful fractal organism, a fractal part of the organism and when you look at this under an electron-microscope, what do you find? Well, it is a bacterium, it is in fact, a rod -shaped bacteria. It has bacillus ??? 16S on both forms, on all forms, you find that it is Bacillus Licheniformis. All those beautiful, what look like fungal branching and so on is, in fact, bacterial rods coming together to form these fractals and forming rings and all kinds of things which are very, very interesting to look at.

Now, I hadn’t heard of B. Licheniformis and although I did a literature search on cancer and bacteria I hadn’t come across it but, in fact, if you go back to the 1950s, a woman in Birmingham called Phyllis Peace, did a lot of work on this Bacillus Licheniformis as a cancer agent also as causing numerous disease in humans. This bacterium can go into the erythrocytes and can be a persister, it can hang around and cause all kinds of diseases, arthritis possibly, and all kinds of unusual symptoms. So it is very much like Borrelia but it is a kind of bacillus, nothing unusual, it’s quite straightforward. So that gave me a link.

At least I had found the bacterium that other people had found, and this was from a tumour, and this may well be what is referred to as the historical cancer germ and this is what people have been looking at for the last 50 years. Now, interestingly, this Bacillus Licheniformis is actually prescribed to some people as a pro-biotic and I got an e-mail from a woman in the States who was treated with Bacillus Licheniformis and she came down with the symptoms of ME, or apparently the symptoms of ME. Then I got together with Andy Wright isolating organisms in the blood and we have isolated, in one instance, Bacillus Licheniformis from an ME patient.

Now, if you look here very carefully, you will see that there are some small blobs here which are much smaller. They are what we might call nano-bacteria, very small bacteria. Now this brings us into the next phase of all this mumbo-jumbo and that is what we call filterability. We find that many of these bacteria somehow go through a membrane filter, which is just a couple of bottles on top of each other, made of plastic, sterilised, you suck here, put suction here, it pulls the air through a filter, a membrane filter, and you can have these filters in different sizes. Most micro-biologists use .4 micron, we use .2 or .1 micron, so they are very small holes in the filter and what we find is that some bacteria can go through. If you put media in the bottom here, you find that when you suck it or even sometimes if you don’t suck it, if you let it go through on gravity, something goes through there and contaminates the bottom and it is the same bacterium as the one you put through the top. It is not contamination, you can do 16S on this and prove that this is the same bacteria that goes through.

Now we have done a lot of work trying to find out what the form that goes through is. It could be an L-form, it could be an organism with no cell wall and squeezes through or it could be a very small form, these so-called nano-bacteria forms which may be formed as part of the life cycle, and we can do this with Staph, so maybe Staph has a small form that we have missed. So point 2 is very small (0.2 microns). It is smaller than the original organism by far.

So this brings us to this thing of nano-bacteria, sometimes spelled with two n’s. This traditionally, in my mind, just means very small bacteria but, unfortunately, I have used the word highjacked, it has been highjacked by the Finnish groups, in particular, to talk about small bacteria that mineralise, that produce minerals and a lot of people say that these are not bacteria they are, in fact, just crystals that are reproducing. So, when that comes along, people then say that small bacteria don’t exist. So, I think we’ve got a problem here in terminology. What we should say is that mineralising nano-bacteria don’t exist perhaps but certainly small bacteria exist. If you starve bacteria they go very small, and you can call them nano-bacteria if you wish.

We have even found these, and this is a total digression, but we have even found these in the stratosphere. We have been looking at the micro-biology of the stratosphere 41 kilometers up, 25 miles, and we find that on particles of space dust or stratosphere dust you get very small bacteria. This is one micron, this is about .5 micron, they get smaller than this. We know that they are bacteria because they produce these little fine things called fimbrii which are like hairs, otherwise, you might think this was just a blob of dust. Here it is showing signs of reproducing, dividing. This is a very small bacteria, they occur everywhere, they are not strictly restricted to the conditions.

Another thing which is amazing about bacteria, which most people kind of get surprised about when you tell them, is that they are intracellular. They can get inside the cells, they can even get inside the nucleus. Now bacterium inside the nucleus with access to all of that DNA can do a lot of damage in terms of changing the genetic structure with bacterium one assumes and they are not restricted to the outside of the cell. This is pretty obvious to anyone who looks at blood under a microscope that this is the case.

Now, something was mentioned about microbiologists having problems with diagnostic microbiology in relation to Borrelia, ME, Lyme Disease and all this and I think this is a cultural problem we have, a major cultural problem that we have, in that all modern bacteriologists and modern microbiologists are actually trained mainly as genetic- or molecular- microbiologists. Now, there is nothing wrong, we use molecular techniques, they are extremely useful but one of the problems is that the time you have to take to teach people how to run gels, or do 16S, or whatever, PCR, means that you have less time to teach them basics and you don’t teach them the basics of microbiology that we used to teach. This is quite obvious, we just don’t have the time or the effort and a lot of the basics are being missed even microscopy is just not part of our curriculum for microbiologists any more. They never see organisms, they are often working with organisms. I often think that many of the PhD theses that may be published relating to say E.coli may have been done on Staph, or something, an extreme example. But the student very rarely looks at the organism and even checks if he or she is using the same organism.

This is a big cultural problem and this is going to get worse because people of my generation, the 50s, late 40s, who were brought up on classical and then kind of converted to genome work are OK in the sense that we have the classical background behind us but for the young people coming up it is going to get worse because in the next 20 years these people will not have any basic micro-biology to go back to. They will certainly not look through microscopes and look at blood in the way that people used to. So it is going to get worse.

So, the conclusion then: Bacteria are not simple organisms that we believe them to be in the textbooks, in our lectures in first year or even third year lectures. Pleomorphism is neglected, well we know that, pleomorphism is neglected and is extremely important in relation to disease causation and many of these life cycles, if they exist, need confirming.