Introduction

According to Prof. Nickolson and associates [₁] human being may be considered as one super-organism consisting of only 10% human cells and 90% bacterial cells, viruses, and fungi. This standpoint highly corresponds to our thesis that nature has created normal flora in human blood on the analogy of oral cavity, skin, gastrointestinal tract, and etc.

The goal of this issue is to demonstrate and visualize normal chlamydia-like flora in human blood on colour microphotos and electron microscopic photos because we found no evident information in available literature for the existence of normal microflora in human blood [₂,₃,₄].

Visualization of normal human blood flora consisting of chlamydia-like microorganisms opens one problem associated with hemotransfusion. Physicians must tackle it in the best way in order to prevent further transfusion of donor's micro-organic flora to the recipient's blood.

Material and Methods

Methods and materials for isolation and typification of ELM are enlisted in literature enclosed.

Electron microscopy

Results

Light microscopy

Figure 1 shows the unfamiliar till now microorganisms resembling chlamidiae, included inside the red blood cells after Gram's stain. The sample has been cultured for 30 days at 37°C. In one erythrocyte, a cluster of several EB of presumable chlamydiae can be seen like in a nest.

Figure 1

Figure 1: CLM inside the human erythrocites under Oil Immersion

Figure 2 shows the EB of presumable chlamydiae outside the erythrocytes because of their destruction with distilled water before culturing. The culture is estimated after 72 hours only because a huge quantity of vitamin K₃ as growth factor 1 g/L was added [₅].

Figure 2

Figure 2: CLM outside the human erythrocites under Oil immersion

Electron microscopy

On Figure 3 and Figure 4, two cells of Mycobacterium tuberculosis ₆ are shown for comparison between the classic microorganism consisting of a multi-layer cellular membrane and a nucleus, and cells from the new microorganisms which have neither a multi-layer cellular membrane wall nor cellular nuclei. Only one fact makes an impression: the new microorganisms are overgrown with tiny and fine pili which are better visualized in younger cells (Figure 5 & 6).

Figure 3

Figure 3: M. tuberculosis - Electron microscopy x 99 000

Figure 4

Figure 4: M. Tuberculosis – Electron microscopy x 46 000

Figure 5

Figure 5: CLM – Electron microscopy x 59 800

Figure 6

Figure 6: CLM – Electron microscopy x 28 800

Reproduction of presumable chlamydiae can be performed either by budding or by simple division of a daughter cell from a mother cell (Figure 7) or by clustering and binding of several divided cells (Figure 8).

Figure 7

Figure 7: CLM budding – Electron microscopy x 34 500

Figure 8

Figure 8: CLM budding – Electron microscopy x 61 000

Figure 9

Figure 9: CLM – Electron microscopy x 41 100

Figure 10

Figure 10: CLM mimicry – Electron microscopy x 9 600

The so-called chlamydiae can be seen inside the erythrocytes as cells with a nucleus-like formation in the central zone surrounded by a light zone with a wall. When found outside the erythrocyte, these cells throw out their nucleus (Figure 9) and remain without a nucleus like erythrocytes themselves. This phenomenon was assumed as mimicry of the new microorganisms, and therefore at the beginning they were called erythrocyte-like microorganisms ELM .

Due to their mimicry, CLM can be seen dispersed among the red blood cells (Figure 10). Discrimination between the two kinds of cells is performed by estimating their dimensions: the new microorganisms are as big as 0.3µm to 2.6µm while the erythrocytes are several times bigger, from 3.5µm to 7.5µm. Much more convincing is the fact that the walls of the new microorganisms are uneven because they are overgrown with multiple tiny and fine pili while the walls of the erythrocytes have no pili and therefore their surface is even.

The presumable chlamydiae have a life cycle. This is a very important argument for the confirmation of the thesis for the existence of chlamydia-like microorganisms. These microorganisms invade and get inside the erythrocyte by passing directly through its wall in different forms due to the fact that the red blood cells have no pili. It is important to mention the fact that, while passing through the erythrocyte wall and being in close contact to it, these microorganisms enlarge their dimensions to a great extent (Figures 11, 12 & 13).

Figure 11

Figure 11: CLM – Electron microscopy x 9 600

Figure 12

Figure 12: CLM – Electron microscopy x 14 300

Figure 13

Figure 13: CLM – Electron microscopy x 27 600

Figure 14

Figure 14: CLM – Electron microscopy x 41 000

Figure 14 demonstrates different forms from the chlamydia-like microorganism's life cycle inside one erythrocyte.

Discussion

All demonstrated microorganisms are isolated from human blood. They cannot be cultivated on microbiologic media which has undergone heat sterilization. Their reproduction becomes possible only if native human blood containing vital erythrocytes is added to the microbiologic media. This is the secret which answers to the question why these CLM are not described in literature till now.

Because of their reproduction only in the presence of vital cells, the new microorganisms are considered intracellular parasites. The availability of a complex life cycle inside and outside the invaded erythrocytes, the formation of cellular forms similar to EB as well as the asymptomatic and long lasting stay in vital human cells are all the landmarks on which a hypothesis can be built that in human erythrocytes live new species called chlamydia-like microorganisms.

Only DNA analysis which is a privilege of scientists in rich countries might answer the question whether a new species (chlamydia-like microorganisms) is found – a fact already predicted in literature [₇].

Conclusions

The discovery of phagocytosis postulated the dogma for the sterility of human blood in healthy individuals.

The discovery of chlamydia-like microorganisms unknown till nowadays which live as normal flora in one part of the erythrocytes would be followed by the proclamation of a new dogma postulating that the blood of every healthy human being is never sterile because a normal flora of chlamydia-like microorganisms lives there.

Disputes between chlamydiologists concerning taxonomic affiliations of the new microorganisms continue for years because of the isolation of one or several more strains. But for the first time chlamydiologists would have enough number of strains to investigate them because everyone would be able to isolate the new microorganisms from its own blood which has already been done by some professors and associates [₈].

It is obvious that millions of patients with surgical interventions and their physicians could by no means wait for years the elucidation of the problem by the chlamydiologists. They cannot wait the elucidation of the unknown microorganisms in order to decide what should be done with donor's blood in which normal flora lives.

The most convenient answer states that, while having in our medical arsenal biotechnological methods for the production of mature erythrocytes [₉], it is not necessary to sterilize donor's blood using irradiation. According to our data, chlamydia-like microorganisms infect erythrocytes produced by stem cells after their influx to the blood pool. That is why the in vitro produced erythrocytes are sterile and suitable for transfusion to the patients without contaminating their blood with the normal flora of the donor

Acknowledgments

Correspondence to

Dr Emil Kalfin 2 Grivitza Str Sofia 1202, Bulgaria Telephone and Fax: + 35928315686 Email: dr_emil_kalfin@yahoo.com