Pseudomonas fluorescens-like bacteria from the stomach:A microbiological and molecular study

AIM:To characterize oxidase-and urease-producing bacterial isolates,grown aerobically,that originated from antral biopsies of patients suffering from acid peptic diseases.METHODS:A total of 258 antral biopsy specimens were subjected to isolation of bacteria followed by tests for oxidase and urease production,acid tolerance and aerobic growth.The selected isolates were further characterized by molecular techniques viz.amplifications for 16S rRNA using universal eubacterial and HSP60 gene specific primers.The amplicons were subjected to restriction analysis and partial sequencing.A phylogenetic tree was generated using unweighted pair group method with arithmetic mean(UPGMA) from evolutionary distance computed with bootstrap test of phylogeny.Assessment of acidity tolerance of bacteria isolated from antrum was performed using hydrochloric acid from 10-7 mol/L to 10-1 mol/L.RESULTS:Of the 258 antral biopsy specimens collected from patients,179(69.4%) were positive for urease production by rapid urease test and 31%(80/258) yielded typical Helicobacter pylori(H.pylori) after 5-7 d of incubation under a microaerophilic environment.A total of 240(93%) antral biopsies yielded homogeneous semi-translucent and small colonies after overnight incubation.The partial 16S rRNA sequences revealed that the isolates had 99% similarity with Pseudomonas species.A phylogenetic tree on the basis of 16S rRNA sequences denoted that JQ927226 and JQ927227 were likely to be related to Pseudomonas fluorescens(P.fluorescens).On the basis ofHSP60 sequences applied to the UPGMA phylogenetic tree,it was observed that isolated strains in an aerobic environment were likely to be P.fluorescens,and HSP60 sequences had more discriminatory potential rather than 16S rRNA sequences.Interestingly,this bacterium was acid tolerant for hours at low pH.Further,a total of 250(96.9%) genomic DNA samples of 258 biopsy specimens and DNA from 240 bacterial isolates were positive for the 613 bp amplicons by targeting P.fluorescens-specific conserved putative outer membrane protein gene sequences.CONCLUSION:This study indicates that bacterial isolates from antral biopsies grown aerobically were P.fluorescens,and thus acid-tolerant bacteria other than H.pylori can also colonize the stomach and may be implicated in pathogenesis/protection.

Thermoresistant, Ethanol-Resistant and Acid-Resistant Properties of Acetic Acid Bacteria Isolated from Fermented Mango Alcohol

Vinegar production is seriously affected by the sensitivity of acetic acid bacteria (AAB) to high temperature, high ethanol concentrations, and high acetic acid concentrations. The aim of this study was to investigate the thermo-ethanol-acid tolerance characters of five AAB strains (VMA1, VMA5, VMA7, VMAM, VMAO) previously isolated from fermented mango alcohol and belonging to Gluconoacetobacter genera. As result, the five AAB strains exhibited good growth and acid production at temperatures up to 45°C;they could tolerate and produce acetic acid at ethanol concentrations up to 20% (v/v). In addition, the studied strains showed growth at acetic acid concentrations up to 4.5% (w/v). Strains VMA7 and VMAO showed the highest resistance properties: they demonstrated acid production at 50°C and VMAO could even grow at 60°C;they tolerated and produced acetic acid at 25% (v/v) ethanol concentration;they showed resistance to acetic acid concentrations up to 6% (w/v). Considering all these properties, the use of these strains would seriously contribute to improving the quality of the vinegar produced and help to reduce the cooling water feeds in vinegar production especially in hot countries in the context of global warming.