Abstract: Nanobacteria are the smallest cell-walled bacteria, only recently discovered in human and cow blood and in commercial cell culture serum. In this study, we identified with energy-dispersive x-ray microanalysis and chemical analysis that all growth phases of nanobacteria produce biogenic apatite on their cell envelope. Fourier transform IR spectroscopy revealed the mineral as carbonate apatite. Previous models for stone formation have lead to a hypothesis that an elevated pH due to urease and/or alkaline phosphatase activity are important lithogenic factors. Our results indicate that carbonate apatite can be formed without these factors at pH 7.4 at physiological phosphate and calcium concentrations. Due to their specific macromolecules, nanobacteria can produce apatite very efficiency in media mimicking tissue fluids and glomerular filtrate and rapidly mineralizing most of available calcium and phosphate. This can be also monitored by $+85$/Sr incorporation and provides a unique model for in vitro studies on calcification. Recently, bacteria have been implicated in the formation of carbonate (hydroxy)fluorapatite in marine sediments. Apatite grains are found so commonly in sedimentary rocks that apatite is omitted in naming the stone. To prove that apatite and other minerals are formed by bacteria would implicate that the bacteria could be observed and their actions followed in stones. We have started to approach this in two ways. Firstly, by the use of sensitive methods for detecting specific bacterial components, like antigens, muramic acid and nucleic acids, that allow for detecting the presence of bacteria and, secondly, by follow-up of volatile bacterial metabolites observed by continuous monitoring with ion mobility spectrometry, IMCELL, working like an artificial, educatable smelling nose. The latter method might allow for remote real time detection of bacterial metabolism, a signature of life, in rocks via fractures of drillholes with or without injected substrate solutions. Nanobacteria may provide a model for primordial life-forms, such as replicating clay crystallites in a sandstone, where minerals and metal atoms associated to membranes, may play catalytic and structural roles reducing the number of enzymes and structural proteins needed for life. Such simple metabolic pathways may support the 10,000-fold slower growth rate of nanobacteria, as compared to the usual bacteria. They may also explain the endurability of this life-form in extreme environmental conditions. Altogether such properties do suggest that nanobacteria may have evolved from environmental sources, such a shot springs, to take advantage of the steady-state calcium and phosphate supply of the mammalian blood. Based upon our findings of nanobacteria, a novel theory for the early development of life, based on apatite-mediated chemistry on membranes selecting itself for its own catalytical machinery, is presented. !26
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