Characterization of bacterial isolates from the microbiota of mothers' breast milk and their infants - PubMed (original) (raw)

Characterization of bacterial isolates from the microbiota of mothers' breast milk and their infants

Kimberly Kozak et al. Gut Microbes. 2015.

Abstract

This investigation assessed the potential of isolating novel probiotics from mothers and their infants. A subset of 21 isolates among 126 unique bacteria from breast milk and infant stools from 15 mother-infant pairs were examined for simulated GI transit survival, adherence to Caco-2 cells, bacteriocin production, and lack of antibiotic resistance. Of the 21 selected isolates a Lactobacillus crispatus isolate and 3 Lactobacillus gasseri isolates demonstrated good profiles of in vitro GI transit tolerance and Caco-2 cell adherence. Bacteriocin production was observed only by L. gasseri and Enterococcus faecalis isolates. Antibiotic resistance was widespread, although not universal, among isolates from infants. Highly similar isolates (≥ 97% similarity by barcode match) of Bifidobacterium longum subsp. infantis (1 match), Lactobacillus fermentum (2 matches), Lactobacillus gasseri (6 matches), and Enterococcus faecalis (1 match) were isolated from 5 infant-mother pairs. Antibiotic resistance profiles between these isolate matches were similar, except in one case where the L. gasseri isolate from the infant exhibited resistance to erythromycin and tetracycline, not observed in matching mother isolate. In a second case, L. gasseri isolates differed in resistance to ampicillin, chloramphenicol and vancomycin between the mother and infant. In this study, gram positive bacteria isolated from mothers' breast milk as well as their infants exhibited diversity in GI transit survival and acid inhibition of pathogens, but demonstrated limited ability to produce bacteriocins. Mothers and their infants offer the potential for identification of probiotics; however, even in the early stages of development, healthy infants contain isolates with antibiotic resistance.

Keywords: antibiotic resistance; bacterial; breast milk; fecal; infant; microbiota; probiotics.

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Figures

Figure 1.

Figure 1.

Example of bacterial barcodes and isolate matching for Lactobacillus isolates from infant-mother pairs. Isolate IDs labeled with 126 are from infant stool and those labeled 124 are from the mother's breast milk.

Figure 2.

Figure 2.

Caco-2 adherence of selected isolates of L. crispatus and L. rhamnosus (Panel A) and L. gasseri (Panel B) from infant stool and breast milk from infant-mother pairs. Reference strains: L. rhamnosus NCK 431 and L. gasseri NCK 99.

Figure 3.

Figure 3.

Bacteriocin screening. Only those isolates, pathogens and indicator combinations for which a zone of inhibition was observed are shown. All other combinations of isolate and pathogens did not result in a zone of inhibition in this spot-overlay detection method. Panels A & B: L. gasseri 291R3 produces a protease (Proteinase K (K) and Pepsin (P)) sensitive bacteriocin against the indicator L. delbeueckii 235_,_ as indicated in Panel A. However, 291 R3's zone of inhibition against E. coli 0157:H7 is NOT sensitive to proteases. (Panel B). Further investigation was required to rule out acid production or an unusual bacteriocin structure. Panel C: Treatments: N- 10 N NaOH (3 μL) E- catalase (3 μL 10 mg/mL) Placing catalase in the zones had NO EFFECT; this ruled out hydrogen peroxide antimicrobial activity. But, placing 3 μL of 10 N NaOH inside the Zones of Inhibition (ZOI), diminished the antimicrobial activity by neutralizing the acid. Therefore, we conclude that the antimicrobial ZOIs against E. coli 0157:H7 are due to acid production. Panel D: The only isolate that produces a bacteriocin against a pathogen is E. faecalis 141 R4. This isolate produces a bacteriocin against Vancomycin Resistant Enterococcus (VRE) and is sensitive to Proteinase K, but not Pepsin (P).

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