Experts On Nanobacteria Face A New Problem: Should Only Synthetic Blood Be Transfused After Normal Flora Has Been Found In Donors' Blood?
E Kalfin
Citation
E Kalfin. Experts On Nanobacteria Face A New Problem: Should Only Synthetic Blood Be Transfused After Normal Flora Has Been Found In Donors' Blood?. The Internet Journal of Microbiology. 2006 Volume 4 Number 1.
Abstract
Contemporary microbiology is deeply engaged in solving the problem of the so-called “nanobacteria”. Transmission electron microscopy (TEM), scanning electron microscopy (SEM) PCR, 16SrRNA analyses, tissue cultures, serologic samples – all these up-to-date microbiologic methods are engaged to reveal nanobacteria's real nature – are they living microorganisms or not? The most competent journals have already tipped the balance toward the standpoint that nanobacteria are self-reproducing calcificating nanoparticles. With regard to this, it is appropriate to use the same modern microbiologic examination methods to prove the findings of normal flora in donors' blood and the consequent necessity of sterile synthetic blood transfusion.
Introduction
Ten expert research teams involved in examining nanobacteria were informed of the Bulgarian discovery of normal flora in the human blood through the Internet, each having received seven publications on this issue (1, 2, 3, 4, 5, 6, 7). Additionally, colour and electron microscopic photos of the bacteria living as normal flora in the blood of the examined Bulgarian professors in medicine were sent. A fast method for the isolation of normal blood flora for 72 hours and a second, accelerated method for the isolation of normal blood flora for 14 days were enclosed, the assessment of results from which was requested by the author.
The author conformed to the fact that seven of these teams suggested the living nature of nanobacteria (8, 9, 10, 11, 12, 13, 14) while the rest three supported the idea of nanobacteria's non-living nature (15, 16, 17).
The author's project related to the exchange of different expert opinions through the Internet aims at evaluating the intellectual capacity of the scientists working with up-to-date microbiological techniques in a situation when a fundamental discovery in microbiology, such as the observation of normal blood flora in human blood, should be estimated.
Materials and Methods
Fast method for isolation of normal blood flora
0.5ml venous blood or 0.5ml blood for transfusion in patients is inoculated in 4.5ml brain heart infusion with 0.25% sodium citrate and 0.1% vitamin K3 (Sigma Aldrich menadione ampoule of 1000mg) and after 72-hour incubation at 43°C in the sediment of the sample, centrifuged for 5 minutes at 3000rev/min, normal blood flora microorganisms, under Gram stain, are visualized.
On sheep blood agar, the subcultures remain sterile but on human blood agar, after 30-day cultivation at 43°C, they form colonies, almost invisible with naked eye. The colonies grow bigger and can be visualized if 1g/L vitamin K3 is added to the human blood agar.
Accelerated method for isolation of normal blood flora
In case of vitamin K3 unavailability, 0.5ml venous blood or 0.5ml blood for transfusion in patients is inoculated in 4.5ml brain heart infusion with 0.25% sodium citrate and the sample is cultured for 14 days at 43°C. The assessment of results follows the described above method. Electron microscopic examination of normal blood flora microorganisms is of great importance since the newly discovered microorganisms have neither visible nucleus nor visible cell wall of Gram-positive or Gram-negative microorganisms; in return, they exhibit a complex developmental cycle described in previous publications.
Results
Fast method for isolation of normal blood flora
The addition of a huge quantity of vitamin K3 (1g/L) as growth factor destroys the erythrocytes; that is why the normal blood flora microorganisms reproduce themselves outside the erythrocytes, in the nutritional medium – being in a free state they grow bigger and start to resemble yeast.
Figure 1
Normal blood flora colonies on human blood agar with vitamin K3 1 g/L after 30 days incubation at 43º C are soft and visible with naked eye
Figure 2
Accelerated method for isolation of normal blood flora
Normal blood flora microorganisms can be seen as a cluster of several spherical bodies inside the human erythrocyte, situated as in a nest. The microorganisms' size is of 0.3µm-2.6µm. More rarely, the microorganisms are located outside the erythrocytes (figure 3).
Figure 3
Discussion
None of the expert research teams involved in examining nanobacteria answered the question “Is human blood sterile?”
Modern scientists cannot or wish not to answer the question, “Is there normal flora in human blood?” Their silence would be appropriate if it turns out that human blood is sterile otherwise it is unjustifiable. The fact that the classic “retro styled” discovery of the microorganisms does not use DNA analysis (as in tuberculosis bacteria case) but a fast 72-hour method for the isolation of normal blood flora culture suggests no excuse for scientists' silence.
Conclusion
The saga related to nanobacteria comes to its end with the recognition of their discoverer, E. O. Kajander (18) that they are non-living self-reproducing nanoparticles. A new saga related to normal flora presence in human blood is coming forth. 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 scientists. Urgent necessity to use modern techniques in order to prove the presence of normal flora in blood exists.
Correspondence to
Dr Emil Kalfin 2 Grivitza Str Sofia 1202, Bulgaria Telephone and Fax : + 35928315686 Email: dr_emil_kalfin@yahoo.com