ISSN:23491604(Volume 2,No.

2,July2015)Reviewarticle

IndexedinSIS(USA),ASI(Germany),I2OR(India)andSJIF(Morocco)databases
ImpactFactor:3.835(SJIF)

Nanobacteria Present Status and Role in Human Diseases

Maninder kaur , Ravikant, Satish gupte, Prerna aggrawal,
Ashwini manahas, Manju arora
Department of Microbiology, Gian Sagar Medical College & Hospital, Punjab. 140601.
*Corresponding author :
Dr. Satish Gupte
Prof.& Head, Department of Microbiology, GianSagar Medical College and Hospital,
Banur, Punjab
e.mail: drsatishgupte@hotmail.com
Manuscript received: 23.03.15
Manuscript accepted: 15.04.15

Abstract
The status of nanobacteria has been controversial, with some researchers suggesting they are a
new class of living organism and others attributing to them a simpler, abiotic nature. They appear
to be ubiquitous entities found in living and non-living substrates. Research has suggested that
nanobacteria could be the cause of a wide variety of diseases, from kidney stones to
atherosclerosis.
Introduction
Nanobacteria are the smallest cell-walled bacteria, discovered in human and cow blood and
commercial cell culture serum. These tiny particles have also been known by the names of
241

SMU Medical Journal, Volume 2, No. 2, July 2015

nanobes and calcifiying nanoparticles[1]. Nanobacteria have intrigued researchers in many ways
since their discovery 20 years ago, but perhaps the most controversial question they pose is
whether or not they are alive. It is debatable if nanobes are living entities [2,3]. Because of their
unique property of biomineralizaton, they have been related to health issues such as the
formation of kidney stones and arterial plaques[4]. Nanobacteria have also been found in
meteorite chipped from the surface of the Red Planet indicated life had once existed on Mars [5].
Geologic research shows they are from the very dawn of life could have shaped Earths early
terrain. They also have been discovered in sedimentary rocks and in hot-springs [1].

Structure and size

According to initial researchers, nanobes are biological structures that are composed of C, O and
N. According to the 16S rRNA gene sequence, nanobacteria fall within the -2 subgroup of
Proteobacteria, which also includes human pathogens like Brucella and Bartonella species [6].
Although they typically have diameters of 0.20.5 m, they can pass through 0.1-m filters
probably because tiny forms (0.050.2 m) are also observed in transmission electron
microscopy (TEM) [4]. Ultra thin sections of nanobes show the existence of an outer layer or
membrane that may represent a cell wall. This outer layer surrounds an electron dense region
interpreted to be the cytoplasm and a less electron dense central region that may represent a
nuclear area. Nanobes show a positive reaction to three DNA stains [49, 6-diamidino-2
phenylindole (DAPI), Acridine Orange and Feulgen] which strongly suggests that nanobes
contain DNA [7].

Controversy
In contrast, recently carried out study by Martel and Young shows that although nanobacterialike particles have a cellular appearance, but they are not alive [8]. Researchers at a workshop
hosted by the National Academy of Sciences for this specific reason concluded that the minimal
cellular size of life on Earth must exceed 200 nm in diameter in order to contain the cellular
machinery based on DNA replication. But nanobacteria can be as small as 80 nm so, unless
242

SMU Medical Journal, Volume 2, No. 2, July 2015

they contain some novel replicating mechanism, it seems unlikely that they constitute a form of
life.8 Moreover, Cisar et al showed that the putative nanobacterial 16SrDNA sequences could
originate from contaminant organism in fetal bovine serum [9].

Present status of nanobacteria

Recent research supports chemical rather than biological model for nanobacteria, being
composed of the calcium phosphate (apatite) covered with organic substances and have been
called nanobacteria-like particles (NLP). The researchers performed a series of experiments
showing that the tiny particles contain no traces of DNA or RNA, and suggest that their
formation

can

be

explained

by

non-biological

means[8].

Unique properties and Koch postulates

Nanobacteria are poorly disruptable, stainable, fixable, and exceptionally resistant to heat. Their
doubling time is about 3 days. High doses of -irradiation or aminoglycoside antibiotics prevent
their multiplication. Nanobacteria possess unusual properties, making their detection difficult
with standard microbiological methods [9]. Another extraordinary property of nanobacteria is
production of carbonate apatite on their cell walls. Nanobacteria may induce calcification and
stone formation in vivo as well as in vitro studies in media mimicking tissue fluids and
glomerular filtrate [4]. Nanobacteria satisfy Koch postulates which further substantiate the link
between nanobacteria and pathologic calcifications. In a small study, Garcia Cuerpo et al found
that translumbar, percutaneous intrarenal injection of NB (isolated from kidney stones) into rats
resulted in kidney stone formation [10]. Another peculiar feature of these bacteria is formation of
slime like biofilms, which may contribute to the stickness of the calcifying nanoparticle complex
in human tissues [11].

Detection methods for nanobacteria

Methods to diagnose NB in biologicals, cells, tissues, blood and urine include immunodetection
with NB-specific monoclonal antibodies, electron microscopy and culture techniques. Because
NB pass through 0.22 m pore size filters, which exclude most common microbes, filtration is
243

SMU Medical Journal, Volume 2, No. 2, July 2015

often used to clean up fluid specimens before culture for NB [12]. Replication can be measured
by particle counting and optical density at 650 nm. It has been also shown that growth of the NB
could be detected by specific methods, such as ELISA, turbidity and SDS-PAGE [1].

NB and Connections to Calcification Related Diseases

NB appears to show a correlation with diverse calcification-related health problems as kidney
stone formation, arterial heart disease, Alzheimers disease, polycystic kidney disease, gall
stones and gallbladder inflammation, prostatitis and cancer. Others have suggested that NB may
contribute to the development of peripheral neutropathy in HIV positive patients and periodontal
problems and even osteoporosis [13].

Role of NB in renal stone formation and mechanism of calcification by nanobacteria

Kidney stones can be composed of a variety of minerals amalgamated with proteins. Regardless
of their mineralogy, two factors are fundamental in kidney stone development: supersaturation
with respect to the forming mineral phase and crystal nucleation. Research has concentrated in
understanding the metabolic and environmental factors which produce an abnormal urinary
composition causing supersaturation [14]. Many studies have indicated nanobacteria as the
nucleating agents of kidney stones, based on the presence of apatite in the core of most kidney
stones, on the widespread occurrence of nanobacteria in kidney stones and on the in vitro
formation of kidney stone-like apatite in the presence of nanobacteria [4,1].

Newer hypothesis in renal stone formation

Now the role of nanobacteria as the nucleating agents is controversial. Aloisi et al suggest that
such particles are non-living and may be formed during the normal living activities of bona-fide
bacteria which inhabit the kidneys [15]. This hypothesis is based on previous observations that
bacteria immersed in a supersaturated fluid produce organic globules which calcify when
released to the surrounding fluid, forming CNP-like particles. These CNP-like particles shared
characteristics with nanobacteria such as small size, an organic nucleus, a calcified mineral shell
244

SMU Medical Journal, Volume 2, No. 2, July 2015

rich in phosphate (detected with Electron Energy Loss Spectroscopy) and the ability to form
colonies [16].Indirect evidence that CNP found in kidneys could be produced by the process
described by Aloisi et al comes from the claim that CNP isolated from kidney stones grow
better in the presence of other bacteria [17].

Role of nanobacteria in vascular calcifications.

Identification of NB-like particles in the lesions produced by vascular calcifications, suggests the
hypothesis that these agents contribute to the pathogenesis of atherosclerosis, especially vascular
calcifications. Specific therapies targeting these particles has demonstrated reduced plaque
formation, regression of plaques, and improved lipid profiles [18]. Epidemiological studies have
implicated antibodies made by the body against nanobacteria to be a strong independent risk
factor for coronary artery calcification, which in turn is an excellent predictor of future coronary
events and death [11].

Role of nanobacteria in calcification in various cancers

Studies have concluded that nanobacteria infection of malignant ovarian tissue contributes to
mechanisms leading to the formation of calcified deposits known as psammoma bodies
[19].They are also known to mediate microcalcifications in breast cancer [11].

Antinanobacterial therapy
Growth could be prevented with tetracycline, high doses of aminoglycoside antibiotics, EDTA,
cytosine arabinoside, 5-FU and gamma-irradiation [1]. Nanobac TX (combination of EDTA and
tetracycline) is safely recommended in the treatment of heart disease [20]. Nanobac OTC
supplement is a proprietary blend of various substances, used in treatment of systemic diseases.
For oral diseases antinanobacterial mouthwashes and toothpastes containing biphosphonates are
recommended [21].

245

SMU Medical Journal, Volume 2, No. 2, July 2015

Conclusion
In the light of such mixed reports, the idea that nanobacteria are living organisms remains highly
controversial, in part because of their very small size and because their nucleic acid is not
sequenced yet. Thus, the focus has momentarily moved on to the calcifying properties of
nanobacteria, rather than on their living or non-living nature .

References
1. Kajander, E.O.; Ciftcioglu, N. (1998). Nanobacteria: An alternative mechanism for
pathogenic intra- and extracellular calcification and stone formation. Proc. Natl. Acad.
Sci. USA, 95(14), 8274-8279.
2. Mckay D N, Mathew G, Kajander E (2006). "Nanobacteria: Fact Or Fiction?
Characteristics, Detection, And Medical Importance Of Novel Self-Replicating,
Calcifying Nanoparticles". J Investig Med 54 (7): 38594
3. Hopkin M.Nanobacteria theory takes a hit.
http://www.nature.com/news/2008/080417/full/news.2008.762
4. Ciftcioglu, N.; Bjorklund, M.; Kuorikoski, K.; Kajander, E.O. (1999).Nanobacteria: an
infectious cause for kidney stone formation. Kidney Int., 56(5), 1893-1898
5. McKay, David S.; et al. (1996). "Search for Past Life on Mars: Possible Relic Biogenic
Activity in Martian Meteorite ALH84001". Science 273 (5277): 924930
6. Ciftcioglu, N., Kuronen, I., kerman, K., Hiltunen, E., Laukkanen, J. & Kajander. E.
O. (1997) in Vaccines 97, eds. Brown, F., Burton, D., Doherty, P., Mekalanos, J. &
Norrby, E. (Cold Spring Harbor Lab. Press, Cold Spring Harbor, NY), pp. 99103.
7. Uwins P J, Webb RI, and Taylor AP.(1998) Novel nano-organisms from Australian
sandstones American Mineralogist, Volume 83, pages 15411550,
8. Martel J, Young JD; Young (April 2008). "Purported nanobacteria in human blood as
calcium carbonate nanoparticles". Proc. Natl. Acad. Sci. U.S.A. 105(14): 554954.

246

SMU Medical Journal, Volume 2, No. 2, July 2015

9. Kajander E O, Tahvanainen E, Kuronen I, iftioglu N (1994) Zentralbl Bakteriol

.Suppl126:147-149
10. Garca-Cuerpo E, Kajander EO, iftiolu N, et al.(2000) Nanobacteria: Un modelo
de neo-litogenesis experimental. Arch. Esp. de Urol., 53:291-303.
11. Miller VM, Rodgers G, Charlesworth JA, Kirkland B, Severson SR, Rasmussen TE,
et al. (2004) Evidence of nanobacterial-like structures in calcified human arteries and
cardiac valves. Am J Physiol Heart Circ Physiol 287:H1115e24.
12. iftioglu N, Kajander EO.(1998) Interaction of Nanobacteria with cultured
mammalian cells. Pathophysiology 4:259-270.
13. An overview on clinical implications of nanobacteria.Kumar CA, Bagga MB, Mohan
V and Raghav N. (2011) J of Indian Academy of oral medicine and radiology,July
23(3)S354-359
14. Moe OW. (2006) Kidney stones: pathophysiology and medical management. Lancet
367:333e44
15 Aloisi G. (2008) An alternative origin for nanobacteria in kidney stones.Bioscience
Hypotheses 1, 138e141
16 Aloisi G, Gloter A, Kruger M, Wallmann K, Guyot F, Zuddas P (2006) Nucleation
of calcium carbonate on bacterial nanoglobules. Geology 34(12):1017e20.
17 Kajander EO. (2006) Nanobacteria-propagating calcifying nanoparticles.Lett Appl
Microbiol. 42:549e52.
18 Puskas LG, Tiszlavicz L, Razga Z, et al. (2005) Detection of nanobacteria-like
particles in human atherosclerotic plaques. Acta Biol Hung 56: 233- 245.
19. Sedivy R, Battistutti WB. (2003) Nanobacteria promote crystallizationof
psammoma bodies in ovarian cancer. AMPIS 111:951e4.
247

SMU Medical Journal, Volume 2, No. 2, July 2015

20. Maniscalco BS, Taylor KA. (2004) Calcification in coronary artery disease can
be reversed by EDTA-tetracycline long-term chemotherapy.
Pathophysiology 11:95-101.
21.Blaizot A, Vergnes JN, Nuwwareh S. (2009) Peri odontal diseases and
cardiovascular events:Meta analysis of observational studies.Int Dent J
59:197-209

Authors Column

SMU Medical Journal, Volume 2, No. 2 , July, 2015, PP. 241 248.
SMU Medical Journal