Isolation, Molecular Identification and Characterization of Bifidobacteria Sp from Breastfed Babies

Abstract

Background: Seventy samples were collected from the feces of healthy newborn babies aged (2-4) weeks and from the arrivals to vaccination centers feed by breast milk and did not take an antibiotic in Wasit province, Iraq. The newborn babies were assessed clinically by the doctors in the hospital, placed directly in the sterile liquid MRS medium in test tubes and transferred to the laboratory within (1-2) hours to complete the isolation processes.

Methods: Bifidobacterium diagnosis was confirmed based on the morphological and cultural characteristics, including size of colonies, color, then studying the characteristics of cells under microscope after staining them with gram stain, additionally biochemical diagnosis of isolates of lactic acid bacteria included catalase test, production of ammonia from arginine hydrolysis, sugars fermentation test, grow in MRS broth, growth at different temperatures, growth at different concentrations of NaCl, resistance test of bacteria to bile salts, determine an ability to lower pH.

Results: that molecular study was done according to bacterial DNA extraction protocol of bacteria using specific primers. The intact DNA bands were confirmed through gel electrophoresis, proving that these isolates are members of the Bifidobacterium genus. The percentage of Bifidobacterium isolates were positive in 6 (12%) and negative in 44 (88%) in collected samples.

Conclusion:

Bifidobacterium is a major genera of bacteria that make up the gastrointestinal tract microbiota in babies, the prevalence of this bacteria is still unknown in babies as the using of antibiotics during pregnancy may affects their number.