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Rational design of improved pharmabiotics.

Sleator RD, Hill C - J. Biomed. Biotechnol. (2009)

Bottom Line: Herein we review the most recent advances in probiotic research and applications with particular emphasis on the novel concept of patho-biotechnology: the application of pathogen-derived (ex vivo and in vivo) stress survival strategies for the design of more technologically robust and effective probiotic cultures with improved biotechnological and clinical applications.

View Article: PubMed Central - PubMed

Affiliation: Alimentary Pharmabiotic Centre, University College Cork, Ireland. roy.sleator@cit.ie

ABSTRACT
Herein we review the most recent advances in probiotic research and applications with particular emphasis on the novel concept of patho-biotechnology: the application of pathogen-derived (ex vivo and in vivo) stress survival strategies for the design of more technologically robust and effective probiotic cultures with improved biotechnological and clinical applications.

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(a) Survival of stationary phase B. breve in 1% porcine bile. B. breve bilE+  (closed circles) and B. breve bilE−  (open circles). Overnight cultures were inoculated (3%) into GM17 and MRS broth containing 1% porcine bile. Viable cell counts were performed by serial dilution in one-quarter strength Ringer's solution followed by plating onto GM17Cm5 or RCMCm4, respectively. Standard deviations of triplicate results are represented by error bars. (b) Effect of bilE on the gastrointestinal persistence of B. breve bilE+  (closed circles) and B. breve bilE−  (open circles) were used for peroral inoculation of female BALB/c mice (n = 5 ). B. breve counts were determined in stools at 48-hour intervals, adapted from Watson et al. [20].
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fig3: (a) Survival of stationary phase B. breve in 1% porcine bile. B. breve bilE+ (closed circles) and B. breve bilE− (open circles). Overnight cultures were inoculated (3%) into GM17 and MRS broth containing 1% porcine bile. Viable cell counts were performed by serial dilution in one-quarter strength Ringer's solution followed by plating onto GM17Cm5 or RCMCm4, respectively. Standard deviations of triplicate results are represented by error bars. (b) Effect of bilE on the gastrointestinal persistence of B. breve bilE+ (closed circles) and B. breve bilE− (open circles) were used for peroral inoculation of female BALB/c mice (n = 5 ). B. breve counts were determined in stools at 48-hour intervals, adapted from Watson et al. [20].

Mentions: Recent work in our laboratory revealed that BetL significantly improved the tolerance of the probiotic strain Bifidobacterium breve UCC2003 to gastric juice [9]. Interestingly, in support of this observation, Termont et al. [21] also reported similar results for an L. lactis strain expressing the E. coli trehalose synthesis genes, thus suggesting a novel protective role for compatible solutes in the gastric environment. Furthermore, inline with our previous observations with L. salivarius UCC118 [8], a significant osmoprotective effect was observed following the introduction of betL into B. breve, allowing significantly improved growth of the probiotic in conditions similar to those encountered in vivo (1.5% NaCl; equivalent to the osmolarity of the gut). In addition, B. breve strains expressing BetL were recovered at significantly higher levels than the wild type in the faeces, intestines, and caecum of inoculated animals. Finally, in addition to improved gastric transit and intestinal persistence (Figure 2(a)), the addition of BetL improved the clinical efficacy of the probiotic culture; mice fed B. breve UCC2003 (betL+) exhibited significantly lower levels of systemic infection compared to the control strain following oral inoculation with L. monocytogenes (Figure 2(b)). Furthermore, in vitro bile tolerance of B. breve was significantly enhanced by heterologous expression of the L. monocytogenes bile resistance mechanism BilE (Figure 3(a)), a phenotype which most likely explains why the bilE+ strain was recovered at significantly higher levels than the control strain from the faeces and intestines of mice, following oral inoculation (Figure 3(b)). In addition, the bilE+ strain demonstrated increased clinical efficacy; by causing a reduction in L. monocytogenes recovered after oral inoculation (Figure 4).


Rational design of improved pharmabiotics.

Sleator RD, Hill C - J. Biomed. Biotechnol. (2009)

(a) Survival of stationary phase B. breve in 1% porcine bile. B. breve bilE+  (closed circles) and B. breve bilE−  (open circles). Overnight cultures were inoculated (3%) into GM17 and MRS broth containing 1% porcine bile. Viable cell counts were performed by serial dilution in one-quarter strength Ringer's solution followed by plating onto GM17Cm5 or RCMCm4, respectively. Standard deviations of triplicate results are represented by error bars. (b) Effect of bilE on the gastrointestinal persistence of B. breve bilE+  (closed circles) and B. breve bilE−  (open circles) were used for peroral inoculation of female BALB/c mice (n = 5 ). B. breve counts were determined in stools at 48-hour intervals, adapted from Watson et al. [20].
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2742647&req=5

fig3: (a) Survival of stationary phase B. breve in 1% porcine bile. B. breve bilE+ (closed circles) and B. breve bilE− (open circles). Overnight cultures were inoculated (3%) into GM17 and MRS broth containing 1% porcine bile. Viable cell counts were performed by serial dilution in one-quarter strength Ringer's solution followed by plating onto GM17Cm5 or RCMCm4, respectively. Standard deviations of triplicate results are represented by error bars. (b) Effect of bilE on the gastrointestinal persistence of B. breve bilE+ (closed circles) and B. breve bilE− (open circles) were used for peroral inoculation of female BALB/c mice (n = 5 ). B. breve counts were determined in stools at 48-hour intervals, adapted from Watson et al. [20].
Mentions: Recent work in our laboratory revealed that BetL significantly improved the tolerance of the probiotic strain Bifidobacterium breve UCC2003 to gastric juice [9]. Interestingly, in support of this observation, Termont et al. [21] also reported similar results for an L. lactis strain expressing the E. coli trehalose synthesis genes, thus suggesting a novel protective role for compatible solutes in the gastric environment. Furthermore, inline with our previous observations with L. salivarius UCC118 [8], a significant osmoprotective effect was observed following the introduction of betL into B. breve, allowing significantly improved growth of the probiotic in conditions similar to those encountered in vivo (1.5% NaCl; equivalent to the osmolarity of the gut). In addition, B. breve strains expressing BetL were recovered at significantly higher levels than the wild type in the faeces, intestines, and caecum of inoculated animals. Finally, in addition to improved gastric transit and intestinal persistence (Figure 2(a)), the addition of BetL improved the clinical efficacy of the probiotic culture; mice fed B. breve UCC2003 (betL+) exhibited significantly lower levels of systemic infection compared to the control strain following oral inoculation with L. monocytogenes (Figure 2(b)). Furthermore, in vitro bile tolerance of B. breve was significantly enhanced by heterologous expression of the L. monocytogenes bile resistance mechanism BilE (Figure 3(a)), a phenotype which most likely explains why the bilE+ strain was recovered at significantly higher levels than the control strain from the faeces and intestines of mice, following oral inoculation (Figure 3(b)). In addition, the bilE+ strain demonstrated increased clinical efficacy; by causing a reduction in L. monocytogenes recovered after oral inoculation (Figure 4).

Bottom Line: Herein we review the most recent advances in probiotic research and applications with particular emphasis on the novel concept of patho-biotechnology: the application of pathogen-derived (ex vivo and in vivo) stress survival strategies for the design of more technologically robust and effective probiotic cultures with improved biotechnological and clinical applications.

View Article: PubMed Central - PubMed

Affiliation: Alimentary Pharmabiotic Centre, University College Cork, Ireland. roy.sleator@cit.ie

ABSTRACT
Herein we review the most recent advances in probiotic research and applications with particular emphasis on the novel concept of patho-biotechnology: the application of pathogen-derived (ex vivo and in vivo) stress survival strategies for the design of more technologically robust and effective probiotic cultures with improved biotechnological and clinical applications.

Show MeSH