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We recently isolated and discovered new  Bradyrhizobiaceae  microbes from the cryopreserved culture broth of blood samples from 3 patients with poorly defined illnesses using modified SP4 media and culture conditions coupled with genomic sequencing. Using a similar protocol, we studied a previously cryopreserved culture broth of blood sample from a patient who had succumbed to an acute onset of fulminant pulmonary illness. We report that two phases of microbial growth were observed in the re-initiated culture. Biochemical and genomic characterization revealed microbes isolated from the first phase of growth were new  Afipia  species of  Bradyrhizobiaceae , tentatively named  A. cberi  with a ~ 5 MB chromosome that was different from those of all previously known  Afipia  microbes including the newly discovered  A. septicemium . The microbes isolated from the second phase of growth were prominent sugar assimilators, novel  Phyllobacteriaceae , phylogenetically most closely related to  Mesorhizobium  and tentatively named  M. hominis  with a ~ 5.5 MB chromosome. All  A. cberi  isolates carry a circular ~ 140 KB plasmid. Some  M. hominis  isolates possess a circular ~ 412 KB plasmid that can be lost in prolonged culture or passage. No antibiotics resistant genes could be identified in both of the  A. cberi  and  M. hominis  plasmids. Antibiotic susceptibility studies using broth culture systems revealed isolates of  A. cberi  could be sensitive to some antibiotics, but all isolates of  M. hominis  were resistant to essentially all tested antibiotics. However, the cell-free antibiotics susceptibility test results may not be applicable to clinical treatment against the microbes that are known to be capable of intracellular growth. It remains to be determined if the 2 previously unknown  Rhizobiales  were indeed pathogenic and played a role in the pulmonary disease process in this patient. Specific probes and methods will be developed to re-examine the diseased lungs from patient's autopsy.

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Isolation and Characterization of Two Novel Bacteria Afipia cberi and Mesorhizobium hominis from Blood of a Patient Afflicted with Fatal Pulmonary Illness