The application of flow cytometry to the characterisation of a probiotic strain Lactobacillus reuteri DPC16 and the evaluation of sugar preservatives for its lyophilization

被引:14
作者
Chen, Song [1 ]
Ferguson, Lynnette R. [2 ]
Shu, Quan [3 ]
Garg, Sanjay [1 ]
机构
[1] Univ Auckland, Auckland Mail Ctr, FMHS, Sch Pharm, Auckland 1142, New Zealand
[2] Univ Auckland, Sch Med Sci, FMHS, Auckland 1142, New Zealand
[3] Bioact Res New Zealand, Auckland, New Zealand
关键词
Probiotics; Flow cytometry; Fluorescent probes; Viability assay; Lyophilization; Lactobacillus reuteri; FRUCTOOLIGOSACCHARIDE UTILIZATION; INDUCED DAMAGE; VIABILITY; SURVIVAL; BACTERIA; OPERON;
D O I
10.1016/j.lwt.2011.05.006
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
In this study, we applied flow cytometric (FCM) analyses to characterize the resistance of a probiotic strain Lactobacillus reuteri DPC16 (DPC16) against diverse stresses. Two fluorescent probes, propidium iodide (PI) and carboxylfluorescein diacetate (cFDA), were combined to the FCM method to reveal multiple cellular statuses of DPC16. The FCM results confirmed that the DPC16 strain had probiotic potential in respect of acid tolerance and bile acid resistance, whereby more than 60% of DPC16 bacteria remained intact after 1 h exposure to pH 2.0, and over half of DPC16 bacteria were unaffected by the presence of bile salts at a concentration of 0.2 g/100 mL for 1 h even without nutrient supply. In addition, the comparison among a number of lyo-preservatives for the DPC16 strain confirmed that lactose was able to maintain over 60% viable DPC16 bacteria after the lyophilization and subsequent storage period, outperforming all the other selected sugars. To conclude, the superior stress resistance of the novel DPC16 strain was confirmed by the FCM analyses in this study. The FCM technique also proved to be readily incorporated into probiotics research, and capable of providing insightful information. (C) 2011 Elsevier Ltd. All rights reserved.
引用
收藏
页码:1873 / 1879
页数:7
相关论文
共 25 条
[1]   Preservation of biological materials under desiccation [J].
Aguilera, JM ;
Karel, M .
CRITICAL REVIEWS IN FOOD SCIENCE AND NUTRITION, 1997, 37 (03) :287-309
[2]   Assessment of high pressure induced damage on Lactobacillus rhamnosus GG by flow cytometry [J].
Ananta, E ;
Heinz, V ;
Knorr, D .
FOOD MICROBIOLOGY, 2004, 21 (05) :567-577
[3]   Functional and comparative genomic analyses of an operon involved in fructooligosaccharide utilization by Lactobacillus acidophilus [J].
Barrangou, R ;
Altermann, E ;
Hutkins, R ;
Cano, R ;
Klaenhammer, TR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (15) :8957-8962
[4]   Multiparametric flow cytometry and cell sorting for the assessment of viable, injured, and dead bifidobacterium cells during bile salt stress [J].
Ben Amor, K ;
Breeuwer, P ;
Verbaarschot, P ;
Rombouts, FM ;
Akkermans, ADL ;
De Vos, WM ;
Abee, T .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2002, 68 (11) :5209-5216
[5]   Intracellular glasses and seed survival in the dry state [J].
Buitink, Julia ;
Leprince, Olivier .
COMPTES RENDUS BIOLOGIES, 2008, 331 (10) :788-795
[6]   Development of a flow cytometric method to analyze subpopulations of bacteria in probiotic products and dairy starters [J].
Bunthof, CJ ;
Abee, T .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2002, 68 (06) :2934-2942
[7]   Flow cytometric assessment of viability of lactic acid bacteria [J].
Bunthof, CJ ;
Bloemen, K ;
Breeuwer, P ;
Rombouts, FM ;
Abee, T .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2001, 67 (05) :2326-2335
[8]   Relevant factors for the preparation of freeze-dried lactic acid bacteria [J].
Carvalho, AS ;
Silva, J ;
Ho, P ;
Teixeira, P ;
Malcata, FX ;
Gibbs, P .
INTERNATIONAL DAIRY JOURNAL, 2004, 14 (10) :835-847
[9]   The role of vitrification in anhydrobiosis [J].
Crowe, JH ;
Carpenter, JF ;
Crowe, LM .
ANNUAL REVIEW OF PHYSIOLOGY, 1998, 60 :73-103
[10]  
CURRY B, 2002, GEN CHARACTERISTICS