Search

Aim: This study aimed to evaluate the protective effect of flaxseed mucilage on the co-extrusion microencapsulation of Lactobacillus rhamnosus GG., Methods: Core flow rate, chitosan coating, and flaxseed mucilage concentration were optimised for the microencapsulation of L. rhamnosus . The microbeads were characterised and evaluated on microencapsulation efficiency and cell released after 6 h of sequential digestion., Results: The optimised parameters for the L. rhamnosus microencapsulation were 1.0 mL/min core flow rate, 0.4% (w/v) chitosan coating, and 0.4% (w/v) flaxseed mucilage. The L. rhamnosus microbeads with flaxseed mucilage in core and wall materials had a smooth surface with 781.3 µm diameter, the highest microencapsulation efficiency (98.8% w/w), lowest swelling (5196.7% w/w) and erosion ratio (515.5% w/w), and least cell release (<40% w/w) with 9.31 log 10  CFU mL -1 after sequential digestion., Conclusions: This study showed the protective capacity of flaxseed mucilage towards the L. rhamnosus GG during microencapsulation and gastrointestinal environment.

Probiotics and curcumin can exhibit synergistic biological activities on the basis of a gut-brain axis, but are sensitive to environmental conditions, making it a challenge for their co-utilization. To meet the demand for high efficiency and convenience, both probiotics and curcumin were encapsulated within a propylene glycol alginate-based hydrogel delivery system, which was assembled using an ethanol-induced approach. The composite hydrogel was effective at sustaining the release of curcumin and protecting LGG cells in simulated gastrointestinal tract conditions. Moreover, it could also largely reduce the chemical degradation of curcumin and increase the survival of LGG during light exposure and long-term storage: up to 91.3 % of curcumin and 9.72 log CFU cm -3 remained present throughout 4 weeks of storage. Results in this work demonstrate a low-energy and green approach to assemble a composite hydrogel with remarkable biocompatibility, which is considered as a desired delivery vehicle for co-delivery of probiotics and curcumin., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Adhesion to cell surfaces is an essential and early prerequisite for successful host colonization by bacteria, and in most instances involves the specificities of various adhesins. Among bacterial Gram-positives, some genera and species mediate attachment to host cells by using long non-flagellar appendages called sortase-dependent pili. A case in point is the beneficial Lactobacillus rhamnosus GG gut-adapted strain that produces the so-called SpaCBA pilus, a structure noted for its promiscuous binding to intestinal mucus and collagen. Structurally, SpaCBA pili are heteropolymers of three different pilin-protein subunits, each with its own location and function in the pilus: backbone SpaA for length, basal SpaB for anchoring, and tip SpaC for adhesion. Previously, we solved the SpaA tertiary structure by X-ray crystallography and also reported on the crystallization of SpaB and SpaC. Here, we reveal the full-length high-resolution (1.9 Å) crystal structure of SpaC, a first for a sortase-dependent pilus-bearing commensal. The SpaC structure, unlike the representative four-domain architecture of other Gram-positive tip pilins, espouses an atypically longer five-domain arrangement that includes N-terminal 'binding' and C-terminal 'stalk' regions of two and three domains, respectively. With the prospect of establishing new mechanistic insights, we provide a structural basis for the multi-substrate binding nature of SpaC, as well as a structural model that reconciles its exclusive localization at the SpaCBA pilus tip., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Introduction: Flow cytometry is a powerful and sensitive technique able to characterize single cells within a heterogeneous population. Different fluorescent dyes can be combined and used together to analyze a great variety of parameters simultaneously. In particular, flow-cytometry allows to measure viability and vitality of probiotics measuring their metabolic activity, fermentation capacity, acidification potential or oxygen uptake ability (Hayouni et al., 2008). To now, plate counting is considered the gold standard in microbiological technique for probiotic enumeration. However, this approach is limited to the detection of only those viable cells which are able to proliferate and form colonies on a solid medium but is not able to recognize not cultivable bacteria and nonviable cells., Aim: The aim of the present study was to apply The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) parameters for the validation of new analytical methods in microbiology. ICH requirements, which are commonly employed for the analysis of drugs and chemical analytes, have been here applied to live cells for the comparison between a flow-cytometric assay and the traditional plate count method for the quantification of viable probiotics bacteria., Methods and Results: Combining specific viability dyes such as thiazole orange (TO) and propidium iodide (PI), probiotic counts of Lactobacillus and Bifidobacterium species were carried out using a FACS Verse (BD Biosciences) cytometer. Analyses were conducted in parallel with the traditional plate count, on specific media. Raw data were analyzed using the FACSuite software (BD Biosciences) and then elaborated with the statistical software Neolicy (VWR International). Results indicated that flow cytometry provides very similar results in cell counting if compared to classical microbiology approaches, showing better performances (ICH parameters) than the traditional plate count method., Conclusions: This work demonstrated the analytical ICH validation of probiotic counts in food supplement products using a robust flow cytometric approach able to enumerate and to assess bacteria viability with stronger results in comparison to the traditional plate count., (Copyright © 2020 Elsevier B.V. All rights reserved.)

To develop a practical food-grade medium and optimal fermentation process for the cost-effective fermentation of Lactobacillus rhamnosus LS-8, both culture medium and conditions were optimized by combining single-factor experimental design, Plackett-Burman design and Box-Behnken design. The medium was simplified to five ingredients (g/L): whey powder (62.5), maltose syrup (50), corn steep liquor (55), NaCl (1) and lysine (0.05), and the optimal culture conditions were initial pH (6.28), constant fermentation pH (4.7), neutralizing agent (NaOH), aeration rate (0.2 v/v/min) and stirrer speed (200 rpm). After culturing in this optimized medium and conditions, the cell density of L. rhamnosus LS-8 was improved to 4.5 × 10 9  CFU/mL, which was elevated about 9 times higher than that obtained in MRS medium. Moreover, cell growth and substrate consumption kinetic constants were determined by the logistic equation and Luedeking-Piret model, and the R 2 values from the model equation were 0.9900 and 0.9971, respectively, indicating that these models were able to simulate the growth and substrate consumption of L. rhamnosus LS-8 accurately. In addition, a high-efficient production process of L. rhamnosus LS-8 was developed by repeated-batch operation, which was verified by five cycles of fermentation with good stability and repeatability. In conclusion, the efficiency of L. rhamnosus LS-8 fermentation was greatly improved as well as the reduction of the cost using the medium and process developed in the present study.

The valorization of a solid carob waste from the Lebanese industry was investigated by optimizing the production of lactic acid using immobilized Lactobacillus rhamnosus in alginate beads and response surface methodology. The results showed that pH and alginate concentration had a significant effect on the production of lactic acid. The fermentation of non-enriched carob waste juice needed an additional nitrogen source to improve lactic acid production and yield. From extracts with 65 g/L sugars, the optimum conditions were found to be 2% for the concentration of alginate, 4% bacteria cells entrapped in beads, 80 rpm agitation speed and pH 6.4. Lactic acid concentration obtained under these conditions was 22 g/L with a yield of 76.9 g/g consumed sugar and a productivity of 1.22 g/L/h. The use of invertase pretreatment increased lactic acid concentration from 22 to 40 g/L, but reduced yield at 66.6%. Finally, cells immobilized in alginate beads could be used for at least five successive cycles., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The polarization imaging technique is a powerful approach to probe microstructural and optical information of biological structures (e.g., tissue samples). Here, we have studied the polarization properties of different bacterial colonies in order to evaluate the possibility of bacterial detection and discrimination. In this regard, we have taken the backscattering Mueller matrix images of four different bacteria colonies (i.e., Escherichia coli, Lactobacillus rhamnosus, Rhodococcus erythropolis, and Staphylococcus aureus). Although the images have the potential to distinguish qualitatively different bacterial colonies, we explored more accurate and quantitative parameters criteria for discrimination of bacterial samples; more specifically, we have exploited the Mueller matrix polar decomposition (MMPD),frequency distribution histogram (FDH), and central moment analysis method. The outcomes demonstrated a superior capacity of Mueller matrix imaging, MMPD, and FDH in bacterial colonies identification and discrimination. This approach might pave the way for a reliable, efficient, and cheap way of identification of infectious diseases.

Lactobacillus rhamnosus DSM 14870 and Lactobacillus gasseri DSM 14869 were previously isolated from the vaginal epithelial cells (VEC) of healthy women and selected for the development of the vaginal EcoVag ® probiotic capsules. EcoVag ® was subsequently shown to provide long-term cure and reduce relapse of bacterial vaginosis (BV) as an adjunct to antibiotic therapy. To identify genes potentially involved in probiotic activity, we performed genome sequencing and characterization of the two strains. The complete genome analysis of both strains revealed the presence of genes encoding functions related to adhesion, exopolysaccharide (EPS) biosynthesis, antimicrobial activity, and CRISPR adaptive immunity but absence of antibiotic resistance genes. Interesting features of L. rhamnosus DSM 14870 genome include the presence of the spaCBA-srtC gene encoding spaCBA pili and interruption of the gene cluster encoding long galactose-rich EPS by integrases. Unique to L. gasseri DSM 14869 genome was the presence of a gene encoding a putative (1456 amino acid) new adhesin containing two rib/alpha-like repeats. L. rhamnosus DSM 14870 and L. gasseri DSM 14869 showed acidification of the culture medium (to pH 3.8) and a strong adhesion capability to the Caco-2 cell line and VEC. L. gasseri DSM 14869 could produce a thick (40nm) EPS layer and hydrogen peroxide. L. rhamnosus DSM 14870 was shown to produce SpaCBA pili and a 20nm EPS layer, and could inhibit the growth of Gardnerella vaginalis, a bacterium commonly associated with BV. The genome sequences provide a basis for further elucidation of the molecular basis for their probiotic functions., (Copyright © 2017 Elsevier GmbH. All rights reserved.)

The goals of this study were to evaluate the precision and accuracy of flow cytometry (FC) methodologies in the evaluation of populations of probiotic bacteria (Lactobacillus rhamnosus R0011) in two commercial dried forms, and ascertain the challenges in enumerating them in a chocolate matrix. FC analyses of total (FC(T)) and viable (FC(V)) counts in liquid or dried cultures were almost two times more precise (reproducible) than traditional direct microscopic counts (DCM) or colony forming units (CFU). With FC, it was possible to ascertain low levels of dead cells (FC(D)) in fresh cultures, which is not possible with traditional CFU and DMC methodologies. There was no interference of chocolate solids on FC counts of probiotics when inoculation was above 10(7) bacteria per g. Addition of probiotics in chocolate at 40 °C resulted in a 37% loss in viable cells. Blending of the probiotic powder into chocolate was not uniform which raised a concern that the precision of viable counts could suffer. FCT data can serve to identify the correct inoculation level of a sample, and viable counts (FCV or CFU) can subsequently be better interpreted., (Crown Copyright © 2014. Published by Elsevier Ltd. All rights reserved.)

The aim of this research was to enhance the survivability of Lactobacillus rhamnosus NRRL 442 against heat exposure via a combination of immobilization and microencapsulation processes using sugarcane bagasse (SB) and sodium alginate (NaA), respectively. The microcapsules were synthesized using different alginate concentration of 1, 2 and 3% and NaA:SB ratio of 1:0, 1:1 and 1:1.5. This beneficial step of probiotic immobilization before microencapsulation significantly enhanced microencapsulation efficiency and cell survivability after heat exposure of 90°C for 30s. Interestingly, the microcapsule of SB-immobilized probiotic could obtain protection from heat using microencapsulation of NaA concentration as low as 1%. SEM images illustrated the incorporation of immobilized L. rhamnosus within alginate matrices and its changes after heat exposure. FTIR spectra confirmed the change in functional bonding in the presence of sugarcane bagasse, probiotic and alginate. The results demonstrated a great potential in the synthesis of heat resistant microcapsules for probiotic., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

We previously described potential probiotic Lactobacillus rhamnosus strains, isolated from fermented mare milk produced in Sumbawa Island, Indonesia, which showed high adhesion to porcine colonic mucin (PCM) and extracellular matrix (ECM) proteins. Recently, mucus-binding factor (MBF) was found in the GG strain of L. rhamnosus as a mucin-binding protein. In this study, we assessed the ability of recombinant MBF protein from the FSMM22 strain, one of the isolates of L. rhamnosus from fermented Sumbawa mare milk, to adhere to PCM and ECM proteins by overlay dot blot and Biacore assays. MBF bound to PCM, laminin, collagen IV, and fibronectin with submicromolar dissociation constants. Adhesion of the FSMM22 mbf mutant strain to PCM and ECM proteins was significantly less than that of the wild-type strain. Collectively, these results suggested that MBF contribute to L. rhamnosus host colonization via mucin and ECM protein binding.

Microbial cells sense and respond to their environment using their surface constituents. Therefore, understanding the assembly and biophysical properties of cell surface molecules is an important research topic. With its ability to observe living microbial cells at nanometer resolution and to manipulate single-cell surface molecules, atomic force microscopy (AFM) has emerged as a powerful tool in microbiology. Here, we survey major breakthroughs made in cell surface microbiology using AFM techniques, emphasizing the most recent structural and functional insights., (Copyright © 2014 Dufrêne.)

In this study, lactic acid and biomass production on liquid distillery stillage from bioethanol production with Lactobacillus rhamnosus ATCC 7469 was studied. The cells were immobilized onto zeolite, a microporous aluminosilicate mineral and the lactic acid production with free and immobilized cells was compared. The immobilization allowed simple cell separation from the fermentation media and their reuse in repeated batch cycles. A number of viable cells of over 10(10) CFU g(-1) of zeolite was achieved at the end of fourth fermentation cycle. A maximal process productivity of 1.69 g L(-1), maximal lactic acid concentration of 42.19 g L(-1) and average yield coefficient of 0.96 g g(-1) were achieved in repeated batch fermentation on the liquid stillage without mineral or nitrogen supplementation., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The aim of this study was to demonstrate the metabolic and proteomic adaptation of Lactobacillus rhamnosus strains, which were isolated at different stages of Parmigiano Reggiano cheese ripening. Compared to de Man, Rogosa, and Sharpe (MRS) broth, cultivation under cheese-like conditions (cheese broth, CB) increased the number of free amino acids used as carbon sources. Compared with growth on MRS or pasteurized and microfiltrated milk, all strains cultivated in CB showed a low synthesis of d,l-lactic acid and elevated levels of acetic acid. The proteomic maps of the five representative strains, showing different metabolic traits, were comparatively determined after growth on MRS and CB media. The amount of intracellular and cell-associated proteins was affected by culture conditions and diversity between strains, depending on their time of isolation. Protein spots showing decreased (62 spots) or increased (59 spot) amounts during growth on CB were identified using MALDI-TOF-MS/MS or LC-nano-ESI-MS/MS. Compared with cultivation on MRS broth, the L. rhamnosus strains cultivated under cheese-like conditions had modified amounts of some proteins responsible for protein biosynthesis, nucleotide, and carbohydrate metabolisms, the glycolysis pathway, proteolytic activity, cell wall, and exopolysaccharide biosynthesis, cell regulation, amino acid, and citrate metabolism, oxidation/reduction processes, and stress responses., (© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

In living cells, sophisticated functional interfaces are generated through the self-assembly of bioactive building blocks. Prominent examples of such biofunctional surfaces are bacterial nanostructures referred to as pili. Although these proteinaceous filaments exhibit remarkable structure and functions, their potential to design bioinspired self-assembled systems has been overlooked. Here, we used atomic force microscopy (AFM) to explore the supramolecular organization and self-assembly of pili from the Gram-positive probiotic bacterium Lactobacillus rhamnosus GG (LGG). High-resolution AFM imaging of cell preparations adsorbed on mica revealed pili not only all around the cells, but also in the form of remarkable star-like structures assembled on the mica surface. Next, we showed that two-step centrifugation is a simple procedure to separate large amounts of pili, even though through their synthesis they are covalently anchored to the cell wall. We also found that the centrifuged pili assemble as long bundles. We suggest that these bundles originate from a complex interplay of mechanical effects (centrifugal force) and biomolecular interactions involving the SpaC cell adhesion pilin subunit (lectin-glycan bonds, hydrophobic bonds). Supporting this view, we found that pili isolated from an LGG mutant lacking hydrophilic exopolysaccharides show an increased tendency to form tight bundles. These experiments demonstrate that AFM is a powerful platform for visualizing individual pili on bacterial surfaces and for unravelling their two-dimensional assembly on solid surfaces. Our data suggest that bacterial pili may provide a generic approach in nanobiotechnology for elaborating functional supramolecular interfaces assembled from bioactive building blocks., (© 2011 American Chemical Society)

Lactobacillus rhamnosus GG, a probiotic with good survival capacity in the human gut, has well-documented adhesion properties and health effects. Recently, spaCBA-encoded pili that bind to human intestinal mucus were identified on its cell surface. Here, we report on the phenotypic analysis of a spaCBA pilus knockout mutant in comparison with the wild type and other adhesin mutants. The SpaCBA pilus of L. rhamnosus GG showed to be key for efficient adherence to the Caco-2 intestinal epithelial cell (IEC) line and biofilm formation. Moreover, the spaCBA mutant induces an elevated level of interleukin-8 (IL-8) mRNA in Caco-2 cells compared to the wild type, possibly involving an interaction of lipoteichoic acid with Toll-like receptor 2. In contrast, an L. rhamnosus GG mutant without exopolysaccharides but with an increased exposure of pili leads to the reduced expression of IL-8. Using Transwells to partition bacteria from Caco-2 cells, IL-8 induction is blocked completely regardless of whether wild-type or mutant L. rhamnosus GG cells are used. Taken together, our data suggest that L. rhamnosus GG SpaCBA pili, while promoting strong adhesive interactions with IECs, have a functional role in balancing IL-8 mRNA expression induced by surface molecules such as lipoteichoic acid.

Aims: Response surface methodology (RSM) was used to optimize a protective medium for enhancing the viability of Lactobacillus rhamnosus E/N cells during lyophilization., Methods and Results: Spirulina, sucrose and lactulose were selected, on the basis of a Plackett-Burman factorial design, as important protectants having the following protective effects on cell viability: 102.025, 36.885 and -34.42, respectively. A full-factorial central composite design was applied to determine optimal levels of three used agents., Conclusion: The optimal protective medium composition was determined to be: Spirulina 1.304% (w/v), lactulose 5.48% (w/v), and sucrose 13.04% (w/v) (Polish Patent P-393189). The predictive value of cell viability in this medium was 89.619%, and experimental viability obtained during freeze-drying was 87.5%., Significance and Impact of the Study: In this study, Spirulina was used for the first time as the protective agent in freeze-drying medium, significantly increasing lactobacilli viability and giving synbiotic character of the final product., (© 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.)

The aim of this study was to demonstrate that flow cytometry (FACS) could potentially be employed for rapid viability assessment of probiotic bacteria immobilized or encapsulated in complex matrices. Lactobacillus rhamnosus GG was immobilized within six different protein environments using whey protein isolate (WPI) and yoghurt matrices and encapsulated within protein micro-beads, all of which ranged in structural complexity. Following a series of environmental-stress trials, survival of the strain was examined using FACS compared to traditional plate count techniques. Cell extraction and digestive pre-treatments were designed to release cells and reduce the protein background, respectively, which represent compositional obstacles for efficient FACS analysis. Physico-chemical properties of protein-probiotic components revealed the mechanism necessary for efficient cell delivery during FACS analysis. This assay required 40 min sample preparation and distinct functional populations were discriminated based on fluorescent properties of thiazole orange (TO) and propidium iodide (PI). This assay yielded 45-50 samples/h, a detection range of 10(2)-10(10)cfu/ml of homogenate and generated correlation coefficients (r) of 0.95, 0.92 and 0.93 in relation to standard plate counts during heat, acid and storage trials, respectively. In conclusion, this methodology provides impetus for dynamic progression of FACS for rapid viability assessment of live bacteria immobilized/encapsulated within complex protein systems., ((c) 2010 Elsevier B.V. All rights reserved.)

This study investigated cell immobilization of Lactobacillus rhamnosus GG in three separate protein products: native, denatured and hydrolysed whey protein isolate (WPI). Treatments were assessed for their ability to enhance probiotic survival during storage, heat stress and ex vivo gastric incubation. Spatial distribution of probiotic cells within immobilized treatments was evaluated by atomic force and confocal scanning laser microscopy, while cell viability was enumerated by plate count and flow cytometry (FACS). Microscopic analysis of denatured treatments revealed an oasis of immobilized cells, phase-separated from the surrounding protein matrix; an environmental characteristic analogous to hydrolysed networks. Cell immobilization in hydrolysed and denatured WPI enhanced survival by 6.1+/-0.1 and 5.8+/-0.1 log10 cycles, respectively, following 14 day storage at 37 degrees C and both treatments generated thermal protection at 57 degrees C (7.3+/-0.1 and 6.5+/-0.1 log(10) cfu/ml). Furthermore, denatured WPI enhanced probiotic protection (8.9+/-0.2 log(10) cfu/ml) following 3h gastric incubation at 37 degrees C. In conclusion, hydrolysed or denatured WPI were the most suitable matrices for cell immobilization, while native protein provided the weakest safeguard against thermal and acid stress, thus making it possible to envision whey protein gel networks as protective substrates for cell immobilization applications., (Copyright 2009 Elsevier B.V. All rights reserved.)

The inactivation of Lb. rhamnosus by pulsed electric field treatment (PEF) was studied in different fractions of raw milk and Ringer solution in order to evaluate the protective effect of nutrient rich media in comparison to aqueous buffer solutions. Apart from monitoring of culturability, analysis of the physiological fitness of Lb. rhamnosus was conducted aiming to identify sublethally damaged cells. Therefore, flow cytometry and a selective medium plating technique were used and compared to each other. The goal of the study was to apply three different parameters describing the physiological fitness of the model organism Lb. rhamnosus after PEF treatment such as culturability, membrane permeability and metabolic activity depending on treatment media and parameters. A concentration dependent protective effect of the milk protein fraction could be shown and allocated to micellar casein as the major milk protein. Increasing the concentration of whey proteins up to 2% showed a similar impact on limiting the PEF inactivation of Lb. rhamnosus. The evaluation of physiological fitness of cells was based on a determination of structural and functional characteristics by rapid cellular staining using carboxyfluorescein diacetate and propidium iodide. This approach showed good accordance to the conventional selective medium plating technique for the enumeration of sublethally-injured bacteria but flow cytometry provided additional information for the characterisation of this fraction. The extent of occurrence of dead, sublethal and vital fractions of cells was found dependent on the PEF treatment parameters such as electrical field strength and energy input as well as the different milk fractions used as treatment media.

The effects of gram-positive bacterial strains (Lactobacillus acidophilus and Lactobacillus rhamnosus) and their subcellular components on the survival of hemopoietic clonogenic cells were evaluated by the formation of endogenous splenic colonies. The effects of these preparations on NO production were studied by the spin-trap paramagnetic resonance spectroscopy. Bacterial preparations from gram-positive bacteria stimulated survival of hemopoietic clonogenic cells, but did not induce NO production in contrast to E. coli LPS.

Objective: We developed a molecular method to mark and differentiate Lactobacillus rhamnosus strains and to analyze genetic diversity among isolates from different sources., Methods: Ten strains of Lactobacillus spp. were isolated from 56 feces samples of elderly people above 90 years of age in regions of Hetian, Xinjiang and Bama, Guangxi, China. We used API 50CHL test chip for strain identification. These isolates belonged to Lactobacillus rhamnosus with 98.6%-99.9% satisfactory. Ten L. rhamnosus isolates and one reference strain ATCC7496 were analyzed by random amplified polymorphic DNA (RAPD) technique to differentiate L. rhamnosus strains at molecular level. We screened 5 random primers named P14, OPG28, OPG25, P7 and P4 and developed optimum RAPD amplifying system., Results: The clear and stable DNA fingerprints of each strain were obtained; the amplicon size ranged from 100 to 2000 bp, and the band patterns showed polymorphism among different L. rhamnosus strains. Genetic similarity coefficients based on RAPD patterns varied from 0.581 to 0.935, which suggested genetic diversity and different genetic relationship among these strains. Eleven strains of Lr could be clustered into 5 groups (group A to E) at the level of 80% similarity. Seven L. rhamnosus strains isolated from Hetian, Xinjiang, were grouped in group B and C, 3 isolates from Bama, Guangxi, were grouped in group D and E., Conclusion: RAPD method is feasible for molecular mark and differentiation of L. rhamnosus strains and high level of genetic diversity and different relationship are found among L. rhamnosus isolates.