Abstract
Intragastric Administration of Lactobacillus plantarum and 2,2'-Dithiodipyridine-Inactivated Simian Immunodeficiency Virus (SIV) Does Not Protect Indian Rhesus Macaques from Intrarectal SIV Challenge or Reduce Virus Replication after Transmission. Diane G Carnathan;Joseph J Mackel;Shelby L Sweat;Chiamaka A Enemuo;Etse H Gebru;Pallavi Dhadvai;Sailaja Gangadhara;Sakeenah Hicks;Thomas H Vanderford;Rama R Amara;José Esparza;Wei Lu;Jean-Marie Andrieu;Guido Silvestri. 2018. J Virol. 92. PMID: 29491157

A major obstacle to development of an effective AIDS vaccine is that along with the intended beneficial responses, the immunization regimen may activate CD4+ T cells that can facilitate acquisition of human immunodeficiency virus (HIV) by serving as target cells for the virus. Lu et al. (W. Lu et al., Cell Rep 2:1736-1746, 2012, https://doi.org/10.1016/j.celrep.2012.11.016) reported that intragastric administration of chemically inactivated simian immunodeficiency virus SIVmac239 and Lactobacillus plantarum (iSIV-L. plantarum) protected 15/16 Chinese-origin rhesus macaques (RMs) from high-dose intrarectal SIVmac239 challenge at 3 months postimmunization. They attributed the observed protection to induction of immune tolerance, mediated by "MHC-Ib/E-restricted CD8+ regulatory T cells that suppressed SIV-harboring CD4+ T cell activation and ex vivo SIV replication in 15/16 animals without inducing SIV-specific antibodies or cytotoxic T." J.-M. Andrieu et al. (Front Immunol 5:297, 2014, https://doi.org/10.3389/fimmu.2014.00297) subsequently reported protection from infection in 23/24 RMs immunized intragastrically or intravaginally with iSIV and Mycobacterium bovis BCG, L. plantarum, or Lactobacillus rhamnosus, which they ascribed to the same tolerogenic mechanism. Using vaccine materials obtained from our coauthors, we conducted an immunization and challenge experiment with 54 Indian RMs and included control groups receiving iSIV only or L. plantarum only as well as unvaccinated animals. Intrarectal challenge with SIVmac239 resulted in rapid infection in all groups of vaccinated RMs as well as unvaccinated controls. iSIV-L. plantarum-vaccinated animals that became SIV infected showed viral loads similar to those observed in animals receiving iSIV only or L. plantarum only or in unvaccinated controls. The protection from SIV transmission conferred by intragastric iSIV-L. plantarum administration reported previously for Chinese-origin RMs was not observed when the same experiment was conducted in a larger cohort of Indian-origin animals.IMPORTANCE Despite an increased understanding of immune responses against HIV, a safe and effective AIDS vaccine is not yet available. One obstacle is that immunization may activate CD4+ T cells that may act as target cells for acquisition of HIV. An alternative strategy may involve induction of a tolerance-inducing response that limits the availability of activated CD4+ T cells, thus limiting the ability of virus to establish infection. In this regard, exciting results were obtained for Chinese-origin rhesus macaques by using a "tolerogenic" vaccine, consisting of intragastric administration of Lactobacillus plantarum and 2,2'-dithiodipyridine-inactivated SIV, which showed highly significant protection from virus transmission. In the present study, we administered iSIV-L. plantarum to Indian-origin rhesus macaques and failed to observe any protective effect on virus acquisition in this experimental setting. This work is important because it contributes to the overall assessment of the clinical potential of a new candidate AIDS vaccine platform based on iSIV-L. plantarum.
Lactobacillus plantarum DSM 2648 is a potential probiotic that enhances intestinal barrier function. Rachel C Anderson;Adrian L Cookson;Warren C McNabb;William J Kelly;Nicole C Roy. 2010. FEMS Microbiol Lett. 309. PMID: 20618863

The aim of this research was to identify bacterial isolates having the potential to improve intestinal barrier function. Lactobacillus plantarum strains and human oral isolates were screened for their ability to enhance tight junction integrity as measured by the transepithelial electrical resistance (TEER) assay. Eight commercially used probiotics were compared to determine which had the greatest positive effect on TEER, and the best-performing probiotic strain, Lactobacillus rhamnosus HN001, was used as a benchmark to evaluate the isolates. One isolate, L. plantarum DSM 2648, was selected for further study because it increased TEER 135% more than L. rhamnosus HN001. The ability of L. plantarum DSM 2648 to tolerate gastrointestinal conditions and adhere to intestinal cells was determined, and L. plantarum DSM 2648 performed better than L. rhamnosus HN001 in all the assays. Lactobacillus plantarum DSM 2648 was able to reduce the negative effect of Escherichia coli [enteropathogenic E. coli (EPEC)] O127:H6 (E2348/69) on TEER and adherence by as much as 98.75% and 80.18%, respectively, during simultaneous or prior coculture compared with EPEC incubation alone. As yet, the precise mechanism associated with the positive effects exerted by L. plantarum DSM 2648 are unknown, and may influence its use to improve human health and wellness.
Ability of Lactobacillus rhamnosus GAF01 to remove AFM1 in vitro and to counteract AFM1 immunotoxicity in vivo. Samir Abbès;Jalila Ben Salah-Abbès;Hakimeh Sharafi;Rania Jebali;Kambiz Akbari Noghabi;Ridha Oueslati. 2012. J Immunotoxicol. 10. PMID: 23030351

Aflatoxin M1 (AFM1) has been detected in many parts of the world both in raw milk and many dairy products, causing great economic losses and human disease. Unfortunately, there are few studies dealing with AFM1 immunotoxicity/interactions with lactic acid bacteria for potential application as a natural preventive agent. The aim of this study was to isolate (from dairy products) food-grade probiotic bacteria able to degrade/bind AFM1 in vitro and evaluate whether the same organism(s) could impart a protective role against AFM1-induced immunotoxicity in exposed Balb/c mice. Bacteria (Lactobacillus plantarum MON03 and L. rhamnosus GAF01) were isolated from Tunisian artisanal butter and then tested for abilities to eliminate AFM1 from phosphate-buffered saline (PBS) and reconstituted milk (containing 0.05, 0.10, and 0.20 µg AFM1/ml) after 0, 6, and 24 h at 37°C. Results showed that the selected bacteria could 'remove' AFM1 both in PBS and skimmed milk. The binding abilities of AFM1 by L. plantarum MON03 and L. rhamnosus GAF01 strains (at 10(8) CFU/ml) in PBS and reconstituted milk ranged, respectively, from 16.1-78.6% and 15.3-95.1%; overall, L. rhamnosus showed a better potential for removal than L. plantarum. 'Removal' appeared to be by simple binding; the bacteria/AFM1 complex was stable and only a very small proportion of mycotoxin was released back into the solution. L. rhamnosus GAF01 had the highest binding capacity and was selected for use in the in vivo study. Those results indicated that use of the organism prevented AFM1-induced effects on total white and red blood cells, and lymphocyte subtypes, after 15 days of host treatment. These studies clearly indicated that L. rhamnosus GAF01 was able to bind AFM1 in vitro and-by mechanisms that might also be related to a binding effect-counteract AFM1-induced immunotoxicity. Moreover, by itself, this bacterium was not toxic and could potentially be used as an additive in dairy products and in biotechnology for mycotoxin detoxification.
Lactobacillus rhamnosus BFE 5264 and Lactobacillus plantarum NR74 Promote Cholesterol Excretion Through the Up-Regulation of ABCG5/8 in Caco-2 Cells. Hong-Sup Yoon;Jae-Hyun Ju;Hannah Kim;Jieun Lee;Hyun-Joon Park;Yosep Ji;Hyeun-Kil Shin;Myoung-Sool Do;Jung-Min Lee;Wilhelm Holzapfel. 2011. Probiotics Antimicrob Proteins. 3. PMID: 26781680

The effect of two putative probiotic strains, Lactobacillus rhamnosus BFE5264 and Lactobacillus plantarum NR74, on the control of cholesterol efflux in enterocytes was assessed by focusing on the promotion of ATP-binding cassette sub-family G members 5 and 8 (ABCG5 and ABCG8). Differentiated Caco-2 enterocytes were treated with live bacteria, heat-killed bacteria, a bacterial cell wall fraction, and metabolites and were subjected to cholesterol uptake assay, mRNA analysis, and protein analyses. Following LXR-transfection by incubation with CHO-K1 cells in DNA-lipofectin added media, the luciferase assay was conducted for LXR analysis. Treatment of Caco-2 cells with L. rhamnosus BFE5264 (isolated from traditional fermented Maasai milk) and L. plantarum NR74 (isolated from Korean kimchi) resulted in the up-regulation of LXR, concomitantly with the elevated expression of ABCG5 and ABCG8. This was associated with the promotion of cholesterol efflux at significantly higher levels compared to the positive control strain L. rhamnosus GG (LGG). The experiment with CHO-K1 cells confirmed up-regulation of LXR-beta by the test strains, and treatment with the live L. rhamnosus BFE5264 and L. plantarum NR74 strains significantly increased cholesterol efflux. Heat-killed cells and cell wall fractions of both LAB strains induced the upregulation of ABCG5/8 through LXR activation. By contrast, LAB metabolites did not show any effect on ABCG5/8 and LXR expression. Data from this study suggest that LAB strains, such as L. rhamnosus BFE5264 and L. plantarum NR74, may promote cholesterol efflux in enterocytes, and thus potentially contribute to the prevention of hypercholesterolemia and atherosclerosis.
The Lactobacillus plantarum strain ACA-DC287 isolated from a Greek cheese demonstrates antagonistic activity in vitro and in vivo against Salmonella enterica serovar Typhimurium. D Fayol-Messaoudi;M-H Coconnier-Polter;V Lievin-Le Moal;F Atassi;C N Berger;A L Servin. 2007. J Appl Microbiol. 103. PMID: 17714399

AIMS: The purpose of this study was to investigate the antibacterial activity of the Xynotyri cheese isolate Lactobacillus plantarum ACA-DC287 using a set of in vitro and in vivo assays. METHODS AND RESULTS: The co-culture of L. plantarum strain ACA-DC287 and Salmonella enterica serovar Typhimurium strain SL1344 results in the killing of the pathogen. The killing activity was produced mainly by non-lactic acid molecule(s) that were present in the cell-free culture supernatant of the L. plantarum strain ACA-DC287. The culture of the L. plantarum strain ACA-DC287 inhibited the penetration of S. typhimurium SL1344 into cultured human enterocyte-like Caco-2/TC7 cells. In conventional mice infected with S. typhimurium SL1344, the intake of L. plantarum strain ACA-DC287 results in a decrease in the levels of Salmonella associated with intestinal tissues or those present in the intestinal contents. In germ-free mice, the L. plantarum strain ACA-DC287 colonized the gastrointestinal tract. CONCLUSIONS: The L. plantarum strain ACA-DC287 strain exerts anti-Salmonella activity similar that of the established probiotic strains Lactobacillus rhamnosus GG, Lactobacillus casei Shirota YIT9029 and Lactobacillus johnsonii La1. SIGNIFICANCE AND IMPACT OF THE STUDY: The observation that a selected cheese Lactobacillus strain exerted antibacterial activity that was similar to those of probiotic Lactobacillus strains, is of interest for the use of this strain as an adjunct strain for the production of health-giving cheeses.
Lactobacillus plantarum CS24.2 prevents Escherichia coli adhesion to HT-29 cells and also down-regulates enteropathogen-induced tumor necrosis factor-α and interleukin-8 expression. Akhilesh S Dhanani;Tamishraha Bagchi. 2013. Microbiol Immunol. 57. PMID: 23586634

The aim of the present study was to evaluate the potential of Lactobacillus plantarum CS24.2 to antagonize Escherichia coli adhesion and modulate expression of the responses by HT-29 cells of inflammatory molecules to E. coli adhesion. Experiments were performed under different adhesion conditions and findings compared with the responses of Lactobacillus rhamnosus GG. Tests of competitive adhesion, adhesion inhibition and displacement assays were performed for lactobacilli (L. rhamnosus GG and L. plantarum CS24.2) and E. coli O26:H11 to HT-29 cells. Both the lactobacilli significantly reduced E. coli adhesion to HT-29 cells (P < 0.05). The ability of lactobacilli to modulate tumor necrosis factor-α and interleukin-8 expression was analyzed in HT-29 cells stimulated with E. coli using qRT-PCR. L. plantarum CS24.2 significantly down regulated expression of both the genes induced by E. coli in HT-29 cells at 6 hr as well as 24 hr, which was more significant than the corresponding findings for L. rhamnosus GG. The present findings suggest that L. plantarum CS24.2 inhibits pathogen adhesion to a similar extent as does the established probiotic strain L. rhamnosus GG. It may also attenuate tumor necrosis factor-α and interleukin-8 expression in HT-29 cells stimulated with E. coli.
Probiotics in foods not containing milk or milk constituents, with special reference to Lactobacillus plantarum 299v. G Molin. 2001. Am J Clin Nutr. 73. PMID: 11157345

Lactic acid fermentation is the simplest and safest way of preserving food and has probably always been used by humans. Species such as Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus acidophilus, and Lactobacillus salivarius are common in the human mucosa, from the mouth to the rectum. In food, L. paracasei and L. rhamnosus are usually associated with dairy products whereas L. plantarum is found in fermented foods of plant origin. A probiotic food product containing no milk constituent was launched in Sweden in 1994. The product is a lactic acid fermented oatmeal gruel that is mixed in a fruit drink. It contains approximately 5 x 10(10) colony-forming units of L. plantarum 299v/L. The strain L. plantarum 299v originates from the human intestinal mucosa and has been shown in rats to decrease translocation, improve mucosal status, improve liver status, improve the immunologic status of the mucosa, and reduce mucosal inflammation. In humans, L. plantarum 299v can increase the concentration of carboxylic acids in feces and decrease abdominal bloating in patients with irritable bowel disease. It can also decrease fibrinogen concentrations in blood. Should probiotics be administrated through foods, the probiotic organism must remain vigorous in the food until consumption and the food must remain palatable, ie, the food carrier and the organism must suit each other. L. plantarum 299v not only affects the bacterial flora of the intestinal mucosa but may also regulate the host's immunologic defense. The mechanisms involved need to be clarified.
Anti-tumour immune effect of oral administration of Lactobacillus plantarum to CT26 tumour-bearing mice. Jingtao Hu;Chunfeng Wang;Liping Ye;Wentao Yang;Haibin Huang;Fei Meng;Shaohua Shi;Zhuang Ding. 2015. J Biosci. 40. PMID: 25963256

Colorectal cancer (CRC) is one of the most prevalent forms of cancer that shows a high mortality and increasing incidence. There are numerous successful treatment options for CRC, including surgery, chemotherapy, radiotherapy and immunotherapy; however, their side effects and limitations are considerable. Probiotics may be an effective strategy for preventing and inhibiting tumour growth through stimulation of host innate and adaptive immunity. We investigated and compared potential anti-tumour immune responses induced by two isolated Lactobacillus strains, Lactobacillus plantarum A and Lactobacillus rhamnosus b, by pre-inoculating mice with lactobacilli for 14 days. Subsequently, subcutaneous and orthotopic intestinal tumours were generated in the pre-inoculated mice using CT26 murine adenocarcinoma cells and were assessed for response against the tumour. Our results indicated that oral administration with L. plantarum inhibited CT26 cell growth in BALB/c mice and prolonged the survival time of tumour-bearing mice compared with mice administered L. rhamnosus. L. plantarum produced protective immunity against the challenge with CT26 cells by increasing the effector functions of CD8+ and natural killer (NK) cell infiltration into tumour tissue, up-regulation of IFN-gamma (but not IL-4 or IL-17) production, and promotion of Th1-type CD4+ T differentiation. Consequently, our results suggest that L. plantarum can enhance the anti-tumour immune response and delay tumour formation.
Transcriptomic responses of Caco-2 cells to Lactobacillus rhamnosus GG and Lactobacillus plantarum J26 against oxidative stress. Yichao Hou;Xuesong Li;Xinyu Liu;Yashuo Zhang;Wei Zhang;Chaoxin Man;Yujun Jiang. 2019. J Dairy Sci. 102. PMID: 31255276

Oxidative stress is the basic reason for aging and age-related diseases. In this study, we investigated the protective effect of 2 strains of lactic acid bacteria (LAB), Lactobacillus rhamnosus GG and L. plantarum J26, against oxidative stress in Caco-2 cells, and gave an overview of the mechanisms of lactic acid bacteria antioxidant activity using digital gene expression profiling. The 2 LAB strains provided significant protection against hydrogen peroxide (H2O2)-induced reduction in superoxide dismutase activity and increase in glutathione peroxidase activity in Caco-2 cells. However, inactive bacteria had little effect on alleviating oxidation stress in Caco-2 cells. Eight genes related to oxidative stress-FOSB, TNF, PPP1R15A, NUAK2, ATF3, TNFAIP3, EGR2, and FBN2-were significantly upregulated in H2O2-induced Caco-2 cells compared with untreated Caco-2 cells. After incubation of the H2O2-induced Caco-2 cells with L. rhamnosus GG and L. plantarum J26, 5 genes (TNF, EGR2, NUAK2, FBN2, and TNFAIP3) and 2 genes (NUAK2 and FBN2) were downregulated, respectively. In addition, the Kyoto Encyclopedia of Genes and Genomes indicated that some signaling pathways associated with inflammation, immune response, and apoptosis, such as Janus kinase/signal transducers and activators of transcription (Jak-STAT), mitogen-activated protein kinase (MAPK), nuclear factor-κB, and tumor necrosis factor, were all negatively modulated by the 2 strains, especially L. rhamnosus GG. In this paper, we reveal the mechanism of LAB in relieving oxidative stress and provide a theoretical basis for the rapid screening and evaluation of new LAB resources.
Eruca sativa might influence the growth, survival under simulated gastrointestinal conditions and some biological features of Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus rhamnosus strains. Florinda Fratianni;Selenia Pepe;Federica Cardinale;Tiziana Granese;Autilia Cozzolino;Raffaele Coppola;Filomena Nazzaro. 2014. Int J Mol Sci. 15. PMID: 25275269

The growth and viability of three Lactobacillus strains, Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus rhamnosus, after their passage through simulated gastric and pancreatic juices were studied as a function of their presence in the growth medium of rocket salad (Eruca sativa). The presence of E. sativa affected some of the biological properties of the strains. For example, L. acidophilus and L. plantarum worked more efficiently in the presence of E. sativa, increasing not only the antioxidant activity of the medium, but also their own antioxidant power and antimicrobial activity; L. rhamnosus was not affected in the same manner. Overall, the presence of vegetables might help to boost, in specific cases, some of the characteristics of lactobacilli, including antioxidant and antimicrobial power.
Detection of the Potential Inactivation of Tetrodotoxin by Lactic Acid Bacterial Exopolysaccharide. Nguyen Hoang Khue Tu;Nghe Van Dat;Le Van Canh;Doan Thi Thanh Vinh. 2018. Toxins (Basel). 10. PMID: 30002293

Screening for compounds that can neutralize the toxicity of tetrodotoxin (TTX) or reduce its negative effects is necessary. Our study tested the TTX detoxification capacity of exopolysaccharide (EPS) extracted from lactic acid bacteria. EPS of Leuconostoc mesenteroides N3 isolated from the Vung Tau sea (Vietnam), Lactobacillus plantarum PN05, and Lactobacillus rhamnosus PN04 were used in the study. To more completely evaluate the importance of EPS in detoxification, EPS samples of Leuconostoc mesenteroides N3, Lactobacillus plantarum PN05 and Lactobacillus rhamnosus PN04 were also tested. The majority of EPS of these bacteria contained glucose; this was observed using thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) analysis. As observed with FTIR analysis, only EPS of Lactobacillus plantarum PN05 contained methyl groups. The results indicated that detoxification of TTX in mice could be obtained at an optimal dose of 248 µg EPS from Leuconostoc mesenteroides incubated with 54 µg cuprous oxide for 40 min or 148 µg EPS Lactobacillus rhamnosus incubated with 55 µg cuprous oxide for 40 min, while EPS from Lactobacillus plantarum showed TTX detoxification capacity without cuprous oxide combination. Consequently, EPS from Lactobacillus plantarum PN05 can be used in TTX prevention. This is the first report on the importance of lactic acid bacteria in TTX detoxification.
Reduction in cholesterol absorption in Caco-2 cells through the down-regulation of Niemann-Pick C1-like 1 by the putative probiotic strains Lactobacillus rhamnosus BFE5264 and Lactobacillus plantarum NR74 from fermented foods. Hong-Sup Yoon;Jae-Hyun Ju;Han-Nah Kim;Hyun-Joon Park;Yosep Ji;Ji-Eun Lee;Hyeun-Kil Shin;Myoung-Sool Do;Wilhelm Holzapfel. 2012. Int J Food Sci Nutr. 64. PMID: 22816655

Hypercholesterolaemia is a major risk factor related to atherosclerosis, and it may be influenced by our diet. This study addresses the impact of Lactobacillus rhamnosus BFE5264 (isolated from Maasai fermented milk) and Lactobacillus plantarum NR74 (from Korean kimchi) on the control of cholesterol absorption through down-regulation of Niemann-Pick C1-like 1 (NPC1L1) expression. Caco-2 enterocytes were treated with the live, heat-killed (HK) bacteria, bacterial cell wall extracts and metabolites; mRNA level and protein expression were measured. Caco-2 cells showed lower NPC1L1 expression in the presence of the live test strains than the control, elucidating down-regulation of cholesterol uptake, and were compared well with the positive control, L. rhamnosus GG. This effect was also observed with HK bacteria and cell wall fractions but not with their metabolites. The potential of some Lactobacillus strains associated with traditional fermented foods to suppress cholesterol uptake and promote its efflux in enterocytes has been suggested from these data.
Distinct immunomodulation of bone marrow-derived dendritic cell responses to Lactobacillus plantarum WCFS1 by two different polysaccharides isolated from Lactobacillus rhamnosus LOCK 0900. Sabina Górska;Martin Schwarzer;Wojciech Jachymek;Dagmar Srutkova;Ewa Brzozowska;Hana Kozakova;Andrzej Gamian. 2014. Appl Environ Microbiol. 80. PMID: 25107979

The structures of polysaccharides (PS) isolated from Lactobacillus rhamnosus LOCK 0900 and results from stimulation of mouse bone marrow-derived dendritic cells (BM-DC) and human embryonal kidney (HEK293) cells stably transfected with Toll-like receptors (TLR) upon exposure to these antigens were studied. L. rhamnosus LOCK 0900 produces PS that differ greatly in their structure. The polymer L900/2, with a high average molecular mass of 830 kDa, is a branched heteropolysaccharide with a unique repeating unit consisting of seven sugar residues and pyruvic acid, whereas L900/3 has a low average molecular mass of 18 kDa and contains a pentasaccharide repeating unit and phosphorus. Furthermore, we found that both described PS neither induce cytokine production and maturation of mouse BM-DC nor induce signaling through TLR2/TLR4 receptors. However, they differ profoundly in their abilities to modulate the BM-DC immune response to the well-characterized human isolate Lactobacillus plantarum WCFS1. Exposure to L900/2 enhanced interleukin-10 (IL-10) production induced by L. plantarum WCFS1, while in contrast, L900/3 enhanced the production of IL-12p70. We conclude that PS, probably due to their chemical features, are able to modulate the immune responses to third-party antigens. The ability to induce regulatory IL-10 by L900/2 opens up the possibility to use this PS in therapy of inflammatory conditions, such as inflammatory bowel disease, whereas L900/3 might be useful in reverting the antigen-dependent Th2-skewed immune responses in allergies.
Prebiotic and synbiotic effects on rats fed malted barley with selected bacteria strains. Yadong Zhong;Margareta Nyman. 2014. Food Nutr Res. 58. PMID: 25317120

BACKGROUND: Butyric acid, one of the key products formed when β-glucans are degraded by the microbiota in the colon, has been proposed to be important for colonic health. Glutamine bound to the fibre may have similar effects once it has been liberated from the fibre in the colon. Both β-glucans and glutamine are found in high amounts in malted barley. Lactobacillus rhamnosus together with malt has been shown to increase the formation of butyric acid further in rats. OBJECTIVE: To investigate whether Lactobacillus rhamnosus 271, Lactobacillus paracasei 87002, Lactobacillus plantarum HEAL 9 and 19, and Bifidobacterium infantis CURE 21 affect the levels of short-chain fatty acids and glutamine in caecum and portal blood of rats fed barley malt. DESIGN: The experimental diets were fed for 12 days. The daily dose of the probiotic strain was 1×10(9) colony forming units and the intake of fibre 0.82 g/day. RESULTS: The malt mostly contained insoluble fibre polymers (93%), consisting of glucose and xylose (38-41 g/kg) and some arabinose (21 g/kg). The fibre polysaccharides were quite resistant to fermentation in the rats, regardless of whether or not probiotics were added (25-30% were fermented). Caecal and portal levels of acetic acid decreased in the rats after the addition of L. plantarum HEAL 9 and L. rhamnosus 271, and also the levels of butyric acid. Viable counts of Lactobacillus, Bifidobacterium and Enterobacteriaceae were unaffected, while the caecal composition of Lactobacilli was influenced by the type of strain administrated. Portal levels of glutamine were unchanged, but glycine levels increased with L. plantarum HEAL 9 and 19 and phenylalanine with L. rhamnosus 271. CONCLUSIONS: Although the probiotic strains survived and reached the caecum, except B. infantis CURE 21, there were no effects on viable counts or in the fermentation of different fibre components, but the formation of some bacterial metabolites decreased. This may be due to the high proportion of insoluble fibres in the malt.
Screening of Lactic Acid Bacteria for Anti-Fusarium Activity and Optimization of Incubation Conditions. Hui Zhao;Anuradha Vegi;Charlene Wolf-Hall. 2017. J Food Prot. 80. PMID: 28853625

Anti-Fusarium activities of lactic acid bacteria (LAB) Lactobacillus plantarum 299V, L. plantarum NRRL-4496, and Lactobacillus rhamnosus VT1 were determined by a microdilution assay developed in this study against Fusarium graminearum 08/RG/BF/51. A cell-free Lactobacillus culture supernatant (CFLCS) of L. rhamnosus VT1 had the highest anti-Fusarium activity. Response surface methodology was used to optimize the incubation conditions for production of CFLCS. A Box-Behnken factorial design was used to investigate the effects of incubation time, shaking speed, and incubation temperature on the inhibition rate of CFLCS. A model equation was generated to predict the inhibition rate of CFLCS under various incubation conditions. A low probability value (0.0012) and associated F value of 25.10 suggested that the model was highly significant. A high R2 value (0.978) indicated a very satisfactory model performance. Response surface methodology analysis suggested that an incubation temperature at 34°C, a shaking speed at 170 rpm, and an incubation time of 55 h were the best combination for production of CFLCS from L. rhamnosus VT1. Under these incubation conditions, a 10% L. rhamnosus VT1 CFLCS solution was predicted to inhibit the growth of F. graminearum by 75.6% in vitro and inhibited 83.7% of the growth in the validation experiment. Thus, the CFLCS of L. rhamnosus VT1 was an effective anti-Fusarium mixture.
Fermentation by Multiple Bacterial Strains Improves the Production of Bioactive Compounds and Antioxidant Activity of Goji Juice. Yuxuan Liu;Huan Cheng;Huiyan Liu;Ruoshuang Ma;Jiangtao Ma;Haitian Fang. 2019. Molecules. 24. PMID: 31569407

Microorganisms can be used for enhancing flavors or metabolizing functional compounds. The fermented-food-derived bacterial strains comprising Bacillus velezensis, Bacillus licheniformis, and Lactobacillus reuteri mixed with Lactobacillus rhamnosus and Lactobacillus plantarum were used to ferment goji berry (Lycium barbarum L.) juice in this study. The fermentation abilities and antioxidant capacities of different mixtures of multiple strains in goji juice were compared. The results showed that the lactic acid contents increased 9.24-16.69 times from 25.30 ± 0.71 mg/100 mL in goji juice fermented using the SLV (Lactobacillus rhamnosus, Lactobacillus reuteri, and Bacillus velezensis), SZP (Lactobacillus rhamnosus, Lactobacillus plantarum, and Bacillus licheniformis), and SZVP (Lactobacillus rhamnosus, Lactobacillus plantarum, Bacillus velezensis, and Bacillus licheniformis) mixtures, and the protein contents increased 1.31-2.11 times from 39.23 ± 0.67 mg/100 mL. In addition, their contents of volatile compounds increased with positive effects on aroma in the fermented juices. Conversion of the free and bound forms of phenolic acids and flavonoids in juice was influenced by fermentation, and the antioxidant capacity improved significantly. Fermentation enhanced the contents of lactic acid, proteins, volatile compounds, and phenols. The antioxidant capacity was strongly correlated with the phenolic composition.
Characterization of a mobile clpL gene from Lactobacillus rhamnosus. Aki Suokko;Kirsi Savijoki;Erja Malinen;Airi Palva;Pekka Varmanen. 2005. Appl Environ Microbiol. 71. PMID: 15812039

Two genes encoding ClpL ATPase proteins were identified in a probiotic Lactobacillus rhamnosus strain, E-97800. Sequence analyses revealed that the genes, designated clpL1 and clpL2, share 80% identity. The clpL2 gene showed the highest degree of identity (98.5%) to a clpL gene from Lactobacillus plantarum WCFSI, while it was not detected in three other L. rhamnosus strains studied. According to Northern analyses, the expression of clpL1 and the clpL2 were induced during heat shock by > 20- and 3-fold, respectively. The functional promoter regions were determined by primer extension analyses, and the clpL1 promoter was found to be overlapped by an inverted repeat structure identical to the conserved CIRCE element, indicating that clpL1 belongs to the HrcA regulon in L. rhamnosus. No consensus binding sites for HrcA or CtsR could be identified in the clpL2 promoter region. Interestingly, the clpL2 gene was found to be surrounded by truncated transposase genes and flanked by inverted repeat structures nearly identical to the terminal repeats of the ISLpl1 from L. plantarum HN38. Furthermore, clpL2 was shown to be mobilized during prolonged cultivation at elevated temperature. The presence of a gene almost identical to clpL2 in L. plantarum and its absence in other L. rhamnosus strains suggest that the L. rhamnosus E-97800 has acquired the clpL2 gene via horizontal transfer. No change in the stress tolerance of the ClpL2-deficient derivative of E-97800 compared to the parental strain was observed.
Acid production and growth by oral Lactobacillus species in vitro. Supatcharin Piwat;Rawee Teanpaisan;Gunnar Dahlén;Songchai Thitasomakul;Charles William Ian Douglas. 2012. J Investig Clin Dent. 3. PMID: 22298522

AIM: To analyze the acid-producing and growth abilities of different oral Lactobacillus species. METHODS: Thirty-nine oral clinical strains and type strains of Lactobacillus, representing nine species, including Lactobacillus casei/paracasei, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus mucosae, Lactobacillus oris, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus salivarius, and Lactobacillus vaginalis were studied. Anaerobically-grown bacterial cells were inoculated overnight in de Man, Rogosa, and Sharpe broth containing 2% glucose (pH 7.0). Acid production and growth were measured at 0, 1.5, 3, 5, 7, and 24 h. RESULTS: Lactobacillus salivarius, Lactobacillus rhamnosus, Lactobacillus casei/paracasei, and Lactobacillus plantarum grew rapidly and reached an optical density higher than other species. They also produced more acid than the others. Lactobacillus vaginalis showed the lowest rate of growth and acid production. These findings demonstrated that the different species of Lactobacillus showed different abilities to generate acid, allowing the species to be categorized into three groups: strongly, moderately, and weakly acidogenic. CONCLUSION: There was variation in acid production and growth between the Lactobacillus species. The strongest acid producers were Lactobacillus salivarius, Lactobacillus rhamnosus, Lactobacillus plantarum, and Lactobacillus casei/paracasei, respectively. It seems possible that these species might play a more important role in caries development than the others.
The probiotic Lactobacillus rhamnosus BFE5264 and Lactobacillus plantarum NR74 promote cholesterol efflux and suppress inflammation in THP-1 cells. Hong-sup Yoon;Jae-hyun Ju;Ji-eun Lee;Hyun-joon Park;Jung-min Lee;Hyeun-kil Shin;Wilhelm Holzapfel;Kun-young Park;Myoung-Sool Do. 2012. J Sci Food Agric. 93. PMID: 22806829

BACKGROUND: The balance between the rate of cholesterol uptake/accumulation and the rate of cholesterol efflux is reflected in the amount of lipid accumulation in macrophages. Based upon the fact that liver X receptors (LXRs) play a role in cholesterol efflux, we studied the effects of probiotics on cholesterol efflux and anti-inflammatory action in macrophages. We confirmed changes in LXR expression by treatment of LXR-transfected CHO-K1 cells with lactic acid bacteria (LAB), and co-cultured THP-1 cells with LAB to investigate changes in cholesterol efflux and inflammation. RESULTS: The experiment with CHO-K1 cells showed upregulation of LXR-β by LAB. Treatment of THP-1 cells with LAB promoted LXR expression in THP-1, which eventually led to significant upregulation of ABCA1 and ABCG1 expression. The treatment with live LAB also significantly promoted cholesterol efflux. LAB suppressed expression of interleukin (IL)-1β and tumor necrosis factor (TNF)-α, which resulted from activation of LXR. CONCLUSION: Our study shows that Lactobacillus rhamnosus BFE5264 and Lactobacillus plantarum NR74 activated LXR and induced cholesterol efflux by promoting expression of ABCA1 and ABCG1. Both strains also suppressed proinflammatory cytokines including IL-1β and TNF-α. This study could account for the observation that LAB may block foam cell formation by cholesterol efflux and immune modulation.
Nutritionally enhanced fermented sausages as a vehicle for potential probiotic lactobacilli delivery. Raquel Rubio;Anna Jofré;Teresa Aymerich;Maria Dolors Guàrdia;Margarita Garriga. 2013. Meat Sci. 96. PMID: 24211552

The suitability of three potential probiotic lactobacilli strains (Lactobacillus casei CTC1677, L. casei CTC1678 and Lactobacillus rhamnosus CTC1679), previously isolated from infants' faeces and characterized, and three commercial probiotic strains (Lactobacillus plantarum 299v, L. rhamnosus GG and L. casei Shirota) was assessed during the manufacture of low-acid fermented sausages (fuets) with reduced Na(+) and fat content. The inoculated strains were successfully monitored by RAPD-PCR during the process. L. rhamnosus CTC1679 was the only strain able to grow and dominate (levels ca. 10(8)CFU/g) the endogenous lactic acid bacteria population in two independent trials, throughout the ripening process. Thus, fuet containing L. rhamnosus CTC1679 as a starter culture could be a suitable vehicle for putative probiotic bacteria delivery. All the final products recorded a satisfactory overall sensory quality without any noticeable off-flavour, and with the characteristic sensory properties of low-acid fermented sausages.
The effect of five probiotic lactobacilli strains on the growth and biofilm formation of Streptococcus mutans. X Lin;X Chen;Y Chen;W Jiang;H Chen. 2014. Oral Dis. 21. PMID: 24806217

OBJECTIVE: To compare the effects of five probiotic lactobacilli strains on the growth and biofilm formation of Streptococcus mutans (MS). MATERIALS AND METHODS: Five probiotic lactobacilli bacteria (LB), Lactobacillus casei Shirota, Lactobacillus casei LC01, Lactobacillus plantarum ST-III, Lactobacillus paracasei Lpc-37, and Lactobacillus rhamnosus HN001, were used as test strains effecting on the Streptococci strain S. mutans UA159 in this study. The effect of LB strains and their supernatants on the viability of the MS was evaluated. Then, the effect of LB strains on the growth of MS biofilm formation was observed by fluorescence microscope. RESULTS: All of the LB strains inhibited the growth of MS at concentrations of 1 × 10(8) and 3 × 10(8) CFU ml(-1) (P < 0.05). Untreated (without pH adjustment and ultrafiltration) LB supernatants from all of the LB strains inhibited the growth of MS (P < 0.05) as well. After pH adjustment and ultrafiltration (treated), only supernatants from L. casei Shirota and L. rhamnosus HN001 inhibited the growth of MS (P < 0.05). MS biofilm formation was also inhibited by all untreated supernatants and by the treated supernatants of L. casei Shirota and L. rhamnosus HN001 (P < 0.05). CONCLUSION: All five probiotic lactobacilli strains inhibited the growth and biofilm formation of MS, likely through the production of an acid environment, bacteriocin-like poly peptides, or both, and the effects on MS were dependent on the LB strains used.
Active Lactobacillus rhamnosus LA68 or Lactobacillus plantarum WCFS1 administration positively influences liver fatty acid composition in mice on a HFD regime. Nevena Ivanovic;Rajna Minic;Ivana Djuricic;Sanja Radojevic Skodric;Irena Zivkovic;Sladjana Sobajic;Brizita Djordjevic. 2016. Food Funct. 7. PMID: 27231730

Western life style, and high calorie diet in particular is causing major health problems such as insulin resistance, hepatic steatosis and heart disease in the modern age. High fat diet (HFD) induces similar changes in mice, such as increased body weight, hypercholesterolemia and accumulation of triglycerides in the liver. These changes can be ameliorated by the administration of some Lactobacillus species. The focus of this study was to analyze the fatty acid content of liver, heart and brain tissues of mice fed HFD and administered with either Lactobacillus plantarum WCFS1 or Lactobacillus rhamnosus LA68, and to analyze the fatty acid content of these organs after a two months washout period. The fatty acid composition of mouse liver tissue changed significantly due to probiotic administration during a 12 weeks HFD regime and active Lactobacillus administration had a slightly reversing effect toward the standard mouse diet group, but after the washout period these changes disappeared. The fatty acid composition of the heart and brain tissues was significantly changed in the HFD regime but probiotic administration had no significant influence on the fatty acid profile of these two organs. Upon the 8 weeks washout period the only remaining beneficial effect was the significantly lower mouse weight in the supplemented groups compared to the HFD group.
Mg2+ improves the thermotolerance of probiotic Lactobacillus rhamnosus GG, Lactobacillus casei Zhang and Lactobacillus plantarum P-8. Y Yang;S Huang;J Wang;G Jan;R Jeantet;X D Chen. 2017. Lett Appl Microbiol. 64. PMID: 28100014

Food-related carbohydrates and proteins are often used as thermoprotectants for probiotic lactobacilli during industrial production and processing. However, the effect of inorganic salts is rarely reported. Magnesium is the second-most abundant cation in bacteria, and commonly found in various foods. Mg2+ homeostasis is important in Salmonella and has been reported to play a critical role in their thermotolerance. However, the role of Mg2+ in thermotolerance of other bacteria, in particular probiotic bacteria, still remains a hypothesis. In this study, the effect of Mg2+ on thermotolerance of probiotic lactobacilli was investigated in three well-documented probiotic strains, Lactobacillus rhamnosus GG, Lactobacillus casei Zhang and Lactobacillus plantarum P-8, in comparison with Zn2+ and Na+ . Concentrations of Mg2+ between 10 and 50 mmol l-1 were found to increase the bacterial survival upon heat challenge. Remarkably, Mg2+ addition at 20 mmol l-1 led to a 100-fold higher survival of L. rhamnosus GG upon heat challenge. This preliminary study also showed that Mg2+ shortened the heat-induced extended lag time of bacteria, which indicated the improvement in bacterial recovery from thermal injury. SIGNIFICANCE AND IMPACT OF THE STUDY: In order to improve the productivity and stability of live probiotics, extensive investigations have been carried out to improve thermotolerance of probiotics. However, most of these studies focused on the effects of carbohydrates, proteins or amino acids. The roles of inorganic salts in various food materials, which have rarely been reported, should be considered when incorporating probiotics into these foods. In this study, Mg2+ was found to play a significant role in the thermotolerance of probiotic lactobacilli. A novel strategy may be available in the near future by employing magnesium salts as protective agents of probiotics during manufacturing process.
Fermented Dessert with Whey, Ingredients from the Peel of Jabuticaba (Myrciaria cauliflora) and an Indigenous Culture of Lactobacillus plantarum: Composition, Microbial Viability, Antioxidant Capacity and Sensory Features. Maria Carmélia Almeida Neta;Anna Paula Rocha de Queiroga;Raphael Lucas Jacinto Almeida;Anderson Caetano Soares;Jade Marinho Gonçalves;Suenia Soares Fernandes;Marina Cínthia de Sousa;Karina Maria Olbrich Dos Santos;Flávia Carolina Alonso Buriti;Eliane Rolim Florentino. 2018. Nutrients. 10. PMID: 30200532

The use of agro-industrial wastes in combination with indigenous lactic acid bacteria is an interesting option to confer functional potential to food products. The microbial viability, chemical composition, antioxidant capacity, texture and sensory acceptability of a fermented dairy dessert containing the indigenous culture Lactobacillus plantarum CNPC003, whey and ingredients obtained from the jabuticaba (Myrciaria cauliflora) peel were compared with formulations without lactobacilli (control) or containing a commercial probiotic culture (Lactobacillus rhamnosus LR32). L. plantarum presented viability higher than 7 log CFU g-1 in the dessert, as did the commercial probiotic, for 21 days at 4 ± 1 °C. Total phenolic contents (45⁻60 mg gallic acid equivalents, GAE, 100 g-1) were comparable to those of other studies evaluating dairy products containing plant sources. The formulations were low in fat, presenting as acceptable for overall consumption, with attractive color and appreciable texture. Considering the total antioxidant capacity, 200⁻250 g of dessert would be necessary to capture 1 g of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals. The dessert with Lactobacillus plantarum CNPC003 is seen as a viable alternative for the use of whey and jabuticaba peel, as well as a potential functional food due to the concentration of lactobacilli reached, besides the presence of antioxidant phenolic compounds.
Short communication: In vitro and in vivo probiotic potential of Lactobacillus plantarum B7 and Lactobacillus rhamnosus D1 isolated from Minas artisanal cheese. G L C Valente;L B Acurcio;L P V Freitas;J R Nicoli;A M Silva;M R Souza;C F A M Penna. 2019. J Dairy Sci. 102. PMID: 31128873

Some Lactobacillus strains may contribute to the health of the host when administered in adequate concentrations, demonstrating their probiotic potential. In contrast, Listeria monocytogenes is a foodborne pathogen that can cause enteropathy, meningoencephalitis, abortion, and septicemia. The aim of this survey was to evaluate the in vitro and in vivo probiotic potential of Lactobacillus plantarum B7 and Lactobacillus rhamnosus D1, isolated from Minas artisanal cheese of the Serra da Canastra (Minas Gerais, Brazil), against Lis. monocytogenes. We submitted B7 and D1 to in vitro testing (antibiogram, tolerance to bile salts and artificial gastric fluid, and spot-on-lawn) and in vivo testing (relative weight gain in mice). Both Lactobacillus strains demonstrated in vitro inhibitory activity against Lis. monocytogenes, as well as sensitivity to antimicrobials and resistance to gastric acids and bile salts. In the in vivo assays, mice treated with D1 gained more weight than mice in the other groups. These results indicate that D1 could have higher probiotic potential than B7 because improvements in feed conversion may help animals fight infection.
Characterization of Lactobacillus rhamnosus MP01 and Lactobacillus plantarum MP02 and Assessment of Their Potential for the Prevention of Gastrointestinal Infections in an Experimental Canine Model. Leónides Fernández;Raquel Martínez;Manuela Pérez;Rebeca Arroyo;Juan M Rodríguez. 2019. Front Microbiol. 10. PMID: 31178838

The aim of the present study was to evaluate the probiotic potential of Lactobacillus rhamnosus MP01 and Lactobacillus plantarum MP02, two strains isolated from canine milk. These two isolates were characterized in vitro for their survival to conditions similar to those found in the canine gastrointestinal tract, production of antimicrobial compounds, adherence to intestinal mucin, degradation of mucin, and antibiotic sensitivity. Globally, both strains exhibited a high in vitro probiotic potential. Finally, their potential for the prevention of gastrointestinal infections was evaluated in an experimental canine model using 1-month-old puppies. A group of 12 German shepherd puppies, 6 males and 6 females, received L. rhamnosus MP01 daily for 2 months and a second group of 12 puppies, 6 males and 6 females, of the same breed received L. plantarum MP02 during the same period of time. The same experimental approach was carried with Yorkshire puppies. Additionally, the trial included 12 dogs of each breed in the placebo groups. The results demonstrated that administration of the strains resulted in a significant preventive effect of gastrointestinal infections in such populations.
Selection of lactic acid bacteria to promote an efficient silage fermentation capable of inhibiting the activity of Aspergillus parasiticus and Fusarium gramineraum and mycotoxin production. C A Dogi;A Fochesato;R Armando;B Pribull;M M S de Souza;I da Silva Coelho;D Araújo de Melo;A Dalcero;L Cavaglieri. 2013. J Appl Microbiol. 114. PMID: 23437822

AIMS: To select lactic acid bacteria with potential silage inoculant properties. The bio-control activity against mycotoxicogenic fungi and the presence of antibiotics resistance gene were also evaluated. METHODS AND RESULTS: Lactobacillus rhamnosus RC007 and Lactobacillus plantarum RC009 were selected on the basis of growth rate and efficacy in reducing the pH of maize extract medium; therefore, they were evaluated for their bio-control ability against Fusarium graminearum and Aspergillus parasiticus. Studies on lag phase, growth rate and aflatoxin B1 (AFB1) and zearalenone (ZEA) production were carried out in vitro under different regimes of aw (0·95 and 0·99); pH (4 and 6); temperature (25 and 37°C); and oxygen availability (normal and reduced). Lactobacillus rhamnosus RC007 was able to completely inhibit the F. graminearum growth at all assayed conditions, while Lact. plantarum RC009 only did it at pH 4. Both Lactobacillus strains were able to significantly reduce the A. parasiticus growth rate mainly at 0·99 aw . A decrease in ZEA production was observed as result of Lactobacillus strains -F. graminearum interaction; however, the A. parasiticus- Lact. plantarum interaction resulted in an increased AFB1 production. Lactobacillus rhamnosus RC007 proved to have no genes for resistance to the tested antibiotics. CONCLUSIONS: The ability of Lact. rhamnosus RC007 to rapidly drop the pH and to inhibit fungal growth and mycotoxin production and the absence of antibiotic resistance genes shows the potential of its application as inoculant and bio-control agent in animal feed. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated the importance of selecting bacteria for silage inoculants not only for the improvement of silage fermentation but also for their effects on mycotoxicogenic fungi and the resulting mycotoxin production due to the risk that they may involve.
The expression of adhesin EF-Tu in response to mucin and its role in Lactobacillus adhesion and competitive inhibition of enteropathogens to mucin. A S Dhanani;T Bagchi. 2013. J Appl Microbiol. 115. PMID: 23663754

AIMS: To analyse the expression of EF-Tu in Lactobacillus strains with response to mucin exposure and its role in interfering with adhesion of enteropathogens to mucin. METHODS AND RESULTS: The Lactobacillus strains were analysed for their ability to adhere to immobilized mucin in microtiter plates. Lactobacillus delbrueckii M and Lactobacillus plantarum CS24.2 showed statistically significant adhesion to mucin, which was similar to Lactobacillus rhamnosus GG, the best binding probiotic strain. Lactobacillus rhamnosus GG, Lact. delbrueckii M, Lact. plantarum CS23 and Lact. plantarum CS24.2 were able to effectively antagonize the adhesion of Escherichia coli and Salmonella enterica serovar Typhi to mucin. In the presence of Lactobacillus adhesin - EF-Tu, the adhesion of Lact. delbrueckii M and the strains of Lact. plantarum to mucin was significantly inhibited. Similarly, EF-Tu also reduced the adhesion of enteropathogens to mucin. Furthermore, the relative fold change in gene expression analysis showed significant up-regulation of EF-Tu gene in the strains of Lact. plantarum and Lact. delbrueckii M when exposed to mucin for 3 h. CONCLUSIONS: The study shows the significant role of EF-Tu in lactobacilli adhesion and enteropathogens inhibition. SIGNIFICANCE AND IMPACT OF THE STUDY: The study suggests EF-Tu as an important factor linked to the Lactobacillus adhesion as well as enteropathogen inhibition. Lactobacillus plantarum CS23 and Lact. plantarum CS24.2 can be used as potential probiotic strains.
Benzoic Acid Production with Respect to Starter Culture and Incubation Temperature during Yogurt Fermentation using Response Surface Methodology. Hyung-Seok Yu;Na-Kyoung Lee;Hye-Lin Jeon;Su Jin Eom;Mi-Young Yoo;Sang-Dong Lim;Hyun-Dong Paik. 2016. Korean J Food Sci Anim Resour. 36. PMID: 27433115

Benzoic acid is occasionally used as a raw material supplement in food products and is sometimes generated during the fermentation process. In this study, the production of naturally occurring yogurt preservatives was investigated for various starter cultures and incubation temperatures, and considered food regulations. Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus plantarum, Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium bifidum, Bifidobacterium infantis, and Bifidobacterium breve were used as yogurt starter cultures in commercial starters. Among these strains, L. rhamnosus and L. paracasei showed the highest production of benzoic acid. Therefore, the use of L. rhamnosus, L. paracasei, S. thermophilus, and different incubation temperatures were examined to optimize benzoic acid production. Response surface methodology (RSM) based on a central composite design was performed for various incubation temperatures (35-44℃) and starter culture inoculum ratios (0-0.04%) in a commercial range of dairy fermentation processes. The optimum conditions were 0.04% L. rhamnosus, 0.01% L. paracasei, 0.02% S. thermophilus, and 38.12℃, and the predicted and estimated concentrations of benzoic acid were 13.31 and 13.94 mg/kg, respectively. These conditions maximized naturally occurring benzoic acid production during the yogurt fermentation process, and the observed production levels satisfied regulatory guidelines for benzoic acid in dairy products.
In vitro adherence of Lactobacillus strains isolated from the vaginas of healthy Iranian women. Elham Mousavi;Manoochehr Makvandi;Ali Teimoori;Angila Ataei;Shokouh Ghafari;Mahin Najafian;Ziba Ourang;Alireza Samarbaf-Zadeh. 2016. J Chin Med Assoc. 79. PMID: 27562422

BACKGROUND: The lactobacilli are a part of the bacterial flora of the human vagina. Detection of normal Lactobacillus species in the vaginas of healthy women in different geographical locations, and evaluation of their specific properties, can aid in the selection of the best species for preventing sexually transmitted diseases in the future. This study was performed to isolate and identify the Lactobacillus species in the vaginas of healthy women and to evaluate the adherence of these lactobacilli to Vero and HeLa cell lines. METHODS: The study included 100 women. Bacteria were isolated from healthy women and purified. Phenotypic and biochemical tests were performed to identify the lactobacilli. The Lactobacillus species were detected by molecular methods using polymerase chain reaction amplification of the full length of the 16S rDNA of the isolated bacteria. Several isolates of each species were then selected to study their adherence to Vero and HeLa cell lines. RESULTS: Among the 50 samples taken from healthy women meeting the inclusion criteria, Lactobacillus species were identified in 33 (66%) samples. Of these lactobacilli, 14 isolates were Lactobacillus crispatus, six (18.2%) were Lactobacillus gasseri, nine (27%) were Lactobacillus rhamnosus, and the rest were either Lactobacillus salivarius (6%) or Lactobacillus plantarum (6%). L. rhamnosus showed the greatest adhesion to the cells when compared to the other tested species. All the lactobacilli isolated in this study showed a smaller capacity for cell adherence when compared with control species. CONCLUSION: L. crispatus, L. rhamnosus, and L. gasseri were the dominant Lactobacillus species in the vaginas of healthy women in Iran. L. rhamnosus attached more readily to the cells than did the other species; therefore, this isolate is a good candidate for further studies on the potential health benefits and application of lactobacilli as probiotics.
Use of Dairy and Plant-Derived Lactobacilli as Starters for Cherry Juice Fermentation. Annalisa Ricci;Martina Cirlini;Antonietta Maoloni;Daniele Del Rio;Luca Calani;Valentina Bernini;Gianni Galaverna;Erasmo Neviani;Camilla Lazzi. 2019. Nutrients. 11. PMID: 30678152

BACKGROUND: Lactic acid bacteria (LAB) exhibit a great biodiversity that can be exploited for different purposes, such as to enhance flavours or metabolize phenolic compounds. In the present study, the use of dairy and plant-derived LAB strains to perform cherry juice fermentation is reported. METHODS: The growth ability of Lactobacillus plantarum, Lactobacillus casei, Lactobacillus paracasei and Lactobacillus rhamnosus was studied in cherry juice. Profiling of sugars, organic acids and volatile compounds was performed by GC-MS (Gas Chromatography-Mass Spectrometry), while the phenolic fraction was characterized using UHPLC (Ultra High Performance Liquid Chromatography) equipped with a linear ion trap-mass spectrometer. RESULTS: Sucrose significantly decreased in all fermented samples as well as malic acid, converted to lactic acid by malolactic fermentation. The total amount of volatile compounds increased. Specifically, propyl acetate, an ester with fruit notes, reached the highest concentration in L. rhamnosus and L. paracasei (dairy strains) fermented juices. Phenolics were extensively metabolized: caffeic acid was converted into dihydrocaffeic acid, p-coumaric acid into 4-ethylphenol and phenyllactic acid was produced. CONCLUSION: Lactic acid fermentation confer fruit notes to the juice and enhance phenyllactic acids, especially employing dairy strains (L. rhamnosus and L. paracasei). The level of dihydrocaffeic acid, a compound with putative biological activity was also increased (in particular with L. plantarum).
The aggregation of human platelets by Lactobacillus species. D W Harty;M Patrikakis;E B Hume;H J Oakey;K W Knox. 1993. J Gen Microbiol. 139. PMID: 8126421

The ability to aggregate human platelets was examined for five Lactobacillus rhamnosus strains and five Lactobacillus paracasei subsp. paracasei strains isolated from patients with infective endocarditis (IE), 25 laboratory isolates from the same two species, and 14 strains from five other oral species, namely Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus oris, Lactobacillus plantarum and Lactobacillus salivarius. Amongst the L. rhamnosus strains, platelets were aggregated by all five IE strains and 8/16 laboratory strains. For the L. paracasei subsp. paracasei strains, the respective numbers were 2/5 and 2/9. Aggregation also occurred with 11/14 strains of the other five species; each species was represented. The optimal ratio of bacteria to platelets for aggregation was approximately 1:1, and there was considerable variation in the lag phase that preceded aggregation, depending on the source of the platelets. Overall, the lag phase varied between 0.25 +/- 0.1 and 20.4 +/- 3.2 min and the percentage aggregation ranged between 70 +/- 2.6 and 104 +/- 13.5%. Confirmation that aggregation was being observed came from studies with five strains on the inhibitory effects of EDTA, dipyridamole, apyrase, imipramine, acetylsalicylic acid and quinacrine. Inhibition of aggregation by L. rhamnosus strains by the peptide arginine-glycine-aspartic acid-serine (RGDS) further indicated a role for fibronectin and/or fibrinogen. Pronase treatment of cells for 1 h and extraction of bacterial surface components with 0.1 M-Tris/HCl (pH 8.5) at 37 degrees C for 1 h stopped aggregation in 8/9 IE strains. Extracted surface proteins (200 micrograms) completely inhibited platelet aggregation by 8/9 of the homologous strains.(ABSTRACT TRUNCATED AT 250 WORDS)
Effects of lactobacilli on yeast-catalyzed ethanol fermentations. N V Narendranath;S H Hynes;K C Thomas;W M Ingledew. 1997. Appl Environ Microbiol. 63. PMID: 9361399

Normal-gravity (22 to 24 degrees Plato) wheat mashes were inoculated with five industrially important strains of lactobacilli at approximately 10(5), approximately 10(6), approximately 10(7), approximately 10(8), and approximately 10(9) CFU/ml in order to study the effects of the lactobacilli on yeast growth and ethanol productivity. Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus #3, Lactobacillus rhamnosus, and Lactobacillus fermentum were used. Controls with yeast cells but no bacterial inoculation and additional treatments with bacteria alone inoculated at approximately 10(7) CFU/ml of mash were included. Decreased ethanol yields were due to the diversion of carbohydrates for bacterial growth and the production of lactic acid. As higher numbers of the bacteria were produced (depending on the strain), 1 to 1.5% (wt/vol) lactic acid resulted in the case of homofermentative organisms. L. fermentum, a heterofermentative organism, produced only 0.5% (wt/vol) lactic acid. When L. plantarum, L. rhamnosus, and L. fermentum were inoculated at approximately 10(6) CFU/ml, an approximately 2% decrease in the final ethanol concentration was observed. Smaller initial numbers (only 10(5) CFU/ml) of L. paracasei or Lactobacillus #3 were sufficient to cause more than 2% decreases in the final ethanol concentrations measured compared to the control. Such effects after an inoculation of only 10(5) CFU/ml may have been due to the higher tolerance to ethanol of the latter two bacteria, to the more rapid adaptation (shorter lag phase) of these two industrial organisms to fermentation conditions, and/or to their more rapid growth and metabolism. When up to 10(9) CFU of bacteria/ml was present in mash, approximately 3.8 to 7.6% reductions in ethanol concentration occurred depending on the strain. Production of lactic acid and a suspected competition with yeast cells for essential growth factors in the fermenting medium were the major reasons for reductions in yeast growth and final ethanol yield when lactic acid bacteria were present.
The evaluation of mixtures of yeast and potato extracts in growth media for biomass production of lactic cultures. H Gaudreau;N Renard;C P Champagne;D Van Horn. 2002. Can J Microbiol. 48. PMID: 12224561

The effectiveness of yeast extracts (YE) and potato extracts (PE) to promote growth of seven lactic cultures was evaluated by automated spectrophotometry (AS). Two aspects of the growth curve were analysed: (1) maximum biomass obtained (using ODmax) and (2) highest specific growth rate mu(max)) Eleven lots from the same PE-manufacturing process were examined for lot-to-lot variability. The ODmax values of three of the seven strains were significantly affected by lot source, but mu(max) was not significantly affected. The growth of bacteria was systematically lower in base medium containing 100% PE than in base medium containing 100% YE for both ODmax or mu(max) data, which could be related to the lower content in nitrogen-based compounds in PE. In AS assays, highest OD values for Lactobacillus casei EQ28, Lactobacillus rhamnosus R-011, Lactobacillus plantarum EQ12, and Streptococcus thermophilus R-083 were obtained with a mixture of PE and YE. Fermentations (2 L) were also carried out to determine the accuracy of AS to predict biomass levels obtained under fermentation trials. In these fermentations, replacement of 50% YE with PE was shown to enable good growth of S. thermophilus. With L. rhamnosus R-011, a high correlation (R2 = 0.95) was found between ODmax data obtained in the AS assays and that of the 2-L bioreactor when the same growth medium was used for both series of fermentations. However, AS was not as efficient when industrial media were used for the bioreactor assays. The relationship was still good for ODmax between AS data and that of the bioreactor data with L. rhamnosus R-011 in industrial LBS medium (R2 = 0.87), but was very poor with the S. thermophilus R-083 on Rosell #43 industrial medium (R2 = 0.33). Since PE cost 40% less than YE, there are strong economic advantages in considering such a partial replacement of YE by PE.
Identification and characterization of the novel LysM domain-containing surface protein Sep from Lactobacillus fermentum BR11 and its use as a peptide fusion partner in Lactobacillus and Lactococcus. Mark S Turner;Louise M Hafner;Terry Walsh;Philip M Giffard. 2004. Appl Environ Microbiol. 70. PMID: 15184172

Examination of supernatant fractions from broth cultures of Lactobacillus fermentum BR11 revealed the presence of a number of proteins, including a 27-kDa protein termed Sep. The amino-terminal sequence of Sep was determined, and the gene encoding it was cloned and sequenced. Sep is a 205-amino-acid protein and contains a 30-amino-acid secretion signal and has overall homology (between 39 and 92% identity) with similarly sized proteins of Lactobacillus reuteri, Enterococcus faecium, Streptococcus pneumoniae, Streptococcus agalactiae, and Lactobacillus plantarum. The carboxy-terminal 81 amino acids of Sep also have strong homology (86% identity) to the carboxy termini of the aggregation-promoting factor (APF) surface proteins of Lactobacillus gasseri and Lactobacillus johnsonii. The mature amino terminus of Sep contains a putative peptidoglycan-binding LysM domain, thereby making it distinct from APF proteins. We have identified a common motif within LysM domains that is shared with carbohydrate binding YG motifs which are found in streptococcal glucan-binding proteins and glucosyltransferases. Sep was investigated as a heterologous peptide expression vector in L. fermentum, Lactobacillus rhamnosus GG and Lactococcus lactis MG1363. Modified Sep containing an amino-terminal six-histidine epitope was found associated with the cells but was largely present in the supernatant in the L. fermentum, L. rhamnosus, and L. lactis hosts. Sep as well as the previously described surface protein BspA were used to express and secrete in L. fermentum or L. rhamnosus a fragment of human E-cadherin, which contains the receptor region for Listeria monocytogenes. This study demonstrates that Sep has potential for heterologous protein expression and export in lactic acid bacteria.
Antibiotic susceptibility profiles of new probiotic Lactobacillus and Bifidobacterium strains. J S Zhou;C J Pillidge;P K Gopal;H S Gill. 2005. Int J Food Microbiol. 98. PMID: 15681048

The antimicrobial susceptibilities and presence of plasmids in four new probiotic lactic acid bacteria (LAB) strains, Lactobacillus rhamnosus HN001 (DR20) HN067, Lactobacillus acidophilus HN017 and Bifidobacterium lactis HN019 (DR10), were determined. Resistance to 18 commonly used antibiotics was assessed by disk diffusion. The three Lactobacillus strains had similar antibiotic susceptibility profiles to those of Lactobacillus plantarum strain HN045 and two commercial probiotic Lactobacillus strains, GG and LA-1. The B. lactis strain HN019 had a similar profile to three commercial probiotic B. lactis strains (Bb12, HN049 and HN098). All 10 strains were sensitive to the Gram-positive spectrum antibiotics erythromycin and novobiocin, the broad-spectrum antibiotics rifampicin, spectinomycin, tetracycline and chloramphenicol and the beta-lactam antibiotics penicillin, ampicillin and cephalothin. By contrast, most strains were resistant to the Gram-negative spectrum antibiotics fusidic acid, nalidixic acid and polymyxin B and the aminoglycosides neomycin, gentamicin, kanamycin and streptomycin. All three L. rhamnosus strains (HN001, HN067 and GG) were resistant to vancomycin and several strains were also resistant to cloxacillin. Of the four new probiotic strains, only L. rhamnosus HN001 contained plasmids; however, a plasmid-free derivative of HN001 had the same antibiotic susceptibility profile as the parent strain.
Antimicrobial susceptibility of non-enterococcal intrinsic glycopeptide-resistant Gram-positive organisms. Carlos Vay;Roxana Cittadini;Claudia Barberis;Carlos Hernán Rodríguez;Herminia Perez Martínez;Fabiana Genero;Angela Famiglietti. 2006. Diagn Microbiol Infect Dis. 57. PMID: 17049799

Non-enterococcal Gram-positive bacteria that are intrinsically vancomycin-resistant have been infrequently isolated in association with serious infections. However, well-documented infections have lately been reported with increasing frequency. Because these organisms may be pathogens, we tested the MICs of 19 antimicrobial agents by the agar dilution method for predicting susceptibility. The activity of these antimicrobial agents was assessed against 28 strains (Lactobacillus rhamnosus, 6; Lactobacillus acidophilus, 1; Lactobacillus casei, 1; Lactobacillus fermentum, 2; Lactobacillus brevis, 1; Lactobacillus plantarum, 1; Weissella confusa, 2; Leuconostoc mesenteroides, 7; Leuconostoc lactis, 4; Pediococcus acidilactici, 2; Pediococcus pentosaceus, 1), isolated from clinical specimens in an Argentinian university hospital from 1997 to 2003. The MICs of penicillin for 67% of the Lactobacillus strains and 100% of the Leuconostoc spp. and Pediococcus spp. strains tested were in the 0.25-2 microg/mL range. Erythromycin was the most active antimicrobial overall. Multiresistance was observed in 2 strains (Lactobacillus rhamnosus, 1; Lactobacillus plantarum, 1).
Bifidobacterium longum PL03, Lactobacillus rhamnosus KL53A, and Lactobacillus plantarum PL02 in the prevention of antibiotic-associated diarrhea in children: a randomized controlled pilot trial. Henryk Szymański;Małgorzata Armańska;Kinga Kowalska-Duplaga;Hania Szajewska. 2008. Digestion. 78. PMID: 18701826

AIM: To determine the efficacy of a combination of Bifidobacterium longum PL03, Lactobacillus rhamnosus KL53A and Lactobacillus plantarum PL02 for the prevention of antibiotic-associated diarrhea in children. METHODS: Seventy-eight children (age: 5 months to 16 years) with otitis media, and/or respiratory tract infections, and/or urinary tract infections were enrolled in a double-blind randomized control trial in which they received standard antibiotic treatment plus a food supplement containing 10(8) colony-forming units of B. longum, L. rhamnosus and L. plantarum (n = 40) or a placebo (n = 38) orally twice daily for the duration of antibiotic treatment. RESULTS: Patients receiving probiotics had a similar rate of diarrhea (> or =3 loose or watery stools/day for > or =48 h occurring during or up to 2 weeks after the antibiotic therapy) as those receiving placebo (relative risk 0.5, 95% CI 0.06-3.5). The mean number of stools per day was significantly lower in the experimental group (mean difference -0.3 stool/day, 95% CI -0.5 to -0.07). No adverse events were reported. CONCLUSION: The administration of the 3 probiotics did not significantly alter the rate of diarrhea, although it reduced the frequency of stools per day. As the overall frequency of diarrhea was surprisingly low, these results should be interpreted with caution.
The innovative potential of Lactobacillus rhamnosus LR06, Lactobacillus pentosus LPS01, Lactobacillus plantarum LP01, and Lactobacillus delbrueckii Subsp. delbrueckii LDD01 to restore the "gastric barrier effect" in patients chronically treated with PPI: a pilot study. Mario Del Piano;Andrea Anderloni;Marco Balzarini;Marco Ballarè;Stefania Carmagnola;Franco Montino;Marco Orsello;Michela Pagliarulo;Roberto Tari;Liliana Soattini;Filomena Sforza;Luca Mogna;Giovanni Mogna. 2012. J Clin Gastroenterol. 46 Suppl. PMID: 22955351

BACKGROUND: Gastroesophageal reflux disease is a very widespread condition. In Europe, it is estimated that about 175 million people suffer from this disease and have to chronically take drugs to increase gastric pH. The proton pump inhibitors (PPIs) such as omeprazole, lansoprazole, and esomeprazole are the most widely used drug typology in this regard. However, the inhibition of normal gastric acid secretion has important side effects, the most important being bacterial overgrowth in the stomach and duodenum with a concentration of >10⁵ viable cells/mL. As a major consequence of this, many harmful or even pathogenic bacteria contained in some foods could survive the gastric transit and colonize either the stomach itself, the duodenum, or the gut, where they could establish acute and even chronic infections with unavoidable consequences for the host's health. In other words, the "gastric barrier effect" is strongly reduced or even disrupted. To date, there are no real strategies to deal with this widespread, although still relatively little known, problem. The aim of this study was to confirm the gastric bacterial overgrowth in long-term PPI consumers and to assess the efficacy of some probiotic bacteria, belonging to both genera Lactobacillus and Bifidobacterium, in the reduction of gastric and duodenal bacterial overgrowth, therefore partially restoring the gastric barrier effect against foodborne pathogenic bacteria. METHODS: For this purpose, probiotics with a strong demonstrated inhibitory activity on gram-negative bacteria, such as Escherichia coli, were tested in a human intervention trial involving a total of 30 subjects treated with PPIs for either 3 to 12 consecutive months (short-term) or >12 consecutive months (long-term). An additional 10 subjects not taking PPIs were enrolled and used as a control group representing the general population. Four selected probiotics Probiotical SpA (Novara, Italy), namely Lactobacillus rhamnosus LR06 (DSM 21981), Lactobacillus pentosus LPS01 (DSM 21980), Lactobacillus plantarum LP01 (LMG P-21021), and Lactobacillus delbrueckii subsp. delbrueckii LDD01 (DSM 22106) were administered for 10 days to 10 subjects treated with PPIs for >12 months (group B). In the 60 mg formulation, N-acetylcysteine was included as well in light of its well-known mechanical effects on bacterial biofilms. Gastroscopies were performed at the beginning of the study (d0) in all the groups (A, B, C, and D) and after 10 days (d10) in group B only; that is, at the end of probiotics intake. The total viable cells and total Lactobacillus were quantified in gastric juice and duodenal brushing material from all subjects. The results were compared among all the groups and with the control subjects (group D) to confirm the bacterial overgrowth. A comparison was made also between d0 and d10 in group B to quantify the efficacy of the 4 probiotics administered for 10 days. Fecal samples were collected from all groups at d0, including subjects not treated with PPIs, and in group B only at d10. Specific bacterial classes, namely enterococci, total coliforms, E. coli, molds, and yeasts were quantified in all fecal specimens. RESULTS: The results collected confirmed the strong bacterial overgrowth in the stomach and duodenum of people treated with PPIs compared with subjects with a normal intragastric acidity. It is also worth noting that the bacterial cell counts in subjects who underwent a long-term treatment with a PPI were greater than the results from subjects taking these drugs for 3 to 12 months. The intake of 4 specific probiotic strains with a marked antagonistic activity towards 5 E. coli bacteria, including the enterohaemorrhagic O157:H7 strain, and an effective amount of N-acetylcysteine (NAC) was able to significantly reduce bacterial overgrowth in long-term PPI-treated subjects. Total lactobacilli represented the major percentage of bacterial counts, thus demonstrating the ability of such bacteria to colonize the stomach and the duodenum, at least temporarily, and to consequently restore the gastric barrier effect. A significant decrease in fecal enterococci, total coliforms, E. coli, molds, and yeasts in subjects treated with PPIs was recorded at the end of probiotics supplementation (d10) compared with baseline (d0) in group B. This is a further confirmation of the barrier effect also exerted at the stomach level. CONCLUSIONS: PPIs are the most widely sold and used drugs in the world. However, the chronic use of these pharmacological molecules exposes the subject to the risk of foodborne infections as most pathogens are able to survive the gastric transit in a condition of significantly decreased acidity.
Strain-dependent augmentation of tight-junction barrier function in human primary epidermal keratinocytes by Lactobacillus and Bifidobacterium lysates. Reshma Sultana;Andrew J McBain;Catherine A O'Neill. 2013. Appl Environ Microbiol. 79. PMID: 23770906

In this study, we investigated whether probiotic lysates can modify the tight-junction function of human primary keratinocytes. The keratinocytes were grown on cell culture inserts and treated with lysates from Bifidobacterium longum, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus fermentum, or Lactobacillus rhamnosus GG. With the exception of L. fermentum (which decreased cell viability), all strains markedly enhanced tight-junction barrier function within 24 h, as assessed by measurements of transepithelial electrical resistance (TEER). However, B. longum and L. rhamnosus GG were the most efficacious, producing dose-dependent increases in resistance that were maintained for 4 days. These increases in TEER correlated with elevated expression of tight-junction protein components. Neutralization of Toll-like receptor 2 abolished both the increase in TEER and expression of tight-junction proteins induced by B. longum, but not L. rhamnosus GG. These data suggest that some bacterial strains increase tight-junction function via modulation of protein components but the different pathways involved may vary depending on the bacterial strain.
Lactobacillus plantarum LB95 impairs the virulence potential of Gram-positive and Gram-negative food-borne pathogens in HT-29 and Vero cell cultures. Virna Dutra;Ana Carla Silva;Paula Cabrita;Cidália Peres;Xavier Malcata;Luisa Brito. 2015. J Med Microbiol. 65. PMID: 26506821

Listeria monocytogenes, Salmonella enterica and verocytotoxigenic Escherichia coli (VTEC) are amongst the most important agents responsible for food outbreaks occurring worldwide. In this work, two Lactobacillus spp. strains (LABs), Lactobacillus plantarum (LB95) and Lactobacillus paraplantarum (LB13), previously isolated from spontaneously fermenting olive brines, and two reference probiotic strains, Lactobacillus casei Shirota and Lactobacillus rhamnosus GG, were investigated for their ability to attenuate the virulence of the aforementioned pathogens using animal cell culture assays. In competitive exclusion assays, the relative percentages of adhesion and invasion of S. enterica subsp. enterica serovar Enteritidis were significantly reduced when the human HT-29 cell line was previously exposed to LB95. The relative percentage of invasion by Listeria monocytogenes was significantly reduced when HT-29 cells were previously exposed to LB95. In the cytotoxicity assays, the cell-free supernatant of the co-culture (CFSC)of VTEC with LB95 accounted for the lowest value obtained amongst the co-cultures of VTEC with LABs, and was significantly lower than the value obtained with the co-culture of VTEC with the two probiotic reference strains. The cytotoxicity of CFSC of VTEC with both LB95 and LB13 exhibited values not significantly different from the cell-free supernatant of the nonpathogenic E. coli B strain. Our results suggested that LB95 may be able to attenuate the virulence of Gram-positive and Gram-negative food-borne pathogens; together with other reported features of these strains, our data reveal their possible use in probiotic foods due to their interesting potential in preventing enteric infections in humans.
Study on the Influence of Tea Extract on Probiotics in Skim Milk: From Probiotics Propagation to Metabolite. Sha Li;Guangyu Gong;Chengjie Ma;Zhenmin Liu;Jie Cai. 2016. J Food Sci. 81. PMID: 27384493

In this study, the influence of tea extract (TE) on the growth of probiotics in skim milk was examined. Lactobacillus plantarum ST-III, Bifidobacterium bifidum Bb02, Lactobacillus acidophilus NCFM, and Lactobacillus rhamnosus GG were used in this study. The introduction of TE in milk significantly stimulated the propagation and acidification of L. rhamnosus GG and L. acidophilus NCFM. The antioxidant capacities and the total free amino acid contents of all fermented milk products were enhanced by the addition of TE; however, there were different antioxidant properties and free amino acid contents of fermented milk samples fermented by different bacteria. With a 9% (w/w) level, the fermentation with L. rhamnosus GG and L. acidophilus NCFM showed larger numbers of viable cells and faster acidifying rates, as well as excellent antioxidant capacity and abundant free amino acids.  The stimulative effects of TE on probiotics can be considered for industrial purposes and has practical implications for commercial applications.
Survival of Probiotics in Hypromellose Capsules with Rice or Potato Maltodextrin Excipient. Jinru Chen;Allison Bechman;Yaa Asantewaa Kafui Klu;Robert D Phillips. 2016. J Food Sci. 81. PMID: 27681165

There is currently no authorized or established therapeutic level/dose of probiotics for proposed health benefits; however, a daily probiotic consumption of 108 to 1010 CFU has been recommended. This study determined the survival of 5 individual probiotic strains, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, and Bifidobacterium lactis, along with a mixture of the 5 strains in hypromellose capsules with rice or potato maltodextrin at 4, 25, and 37 °C for 12 mo. Samples were collected monthly and plated on deMan-Rogosa Sharpe agar with 0.05% l-cysteine hydrochloride. Results showed that samples stored at 4 °C had an average count of 108 to 1011 CFU/g of probiotic cells during the 12 mo period, whereas at 25 °C, L. rhamnosus and L. paracasei had an average counts below 108 CFU/g during the storage period. L. rhamnosus was the most vulnerable strain used in this study, having the least viable counts at all 3 storage temperatures. Probiotics stored in rice maltodextrin, on average, had higher probiotic counts compared to those stored in potato maltodextrin. Study suggests that to provide consumers with 108 to 1010 CFU/d of probiotic cells, robust bacterial strains, suitable carriers, and a storage temperature of 4 °C are required.
Efficacy and safety of probiotic-supplemented triple therapy for eradication of Helicobacter pylori in children: a systematic review and network meta-analysis. Jue-Rong Feng;Fan Wang;Xiao Qiu;Lynne V McFarland;Peng-Fei Chen;Rui Zhou;Jing Liu;Qiu Zhao;Jin Li. 2017. Eur J Clin Pharmacol. 73. PMID: 28681177

AIM: The aim of this study was to identify the best probiotic supplementation in triple therapy for pediatric population with Helicobacter pylori infection. METHODS: Eligible trials were identified by comprehensive searches. Relative risks with 95% confidence intervals and relative ranks with P scores were assessed. RESULTS: Twenty-nine trials (3122 participants) involving 17 probiotic regimens were identified. Compared with placebo, probiotic-supplemented triple therapy significantly increased H. pylori eradication rates (relative ratio (RR) 1.19, 95% CI 1.13-1.25) and reduced the incidence of total side effects (RR 0.49, 95% CI 0.38-0.65). Furthermore, to supplemented triple therapy, Lactobacillus casei was identified the best for H. pylori eradication rates (P score = 0.84), and multi-strain of Lactobacillus acidophilus and Lactobacillus rhamnosus for total side effects (P score = 0.93). As for the subtypes of side effects, multi-strain of Bifidobacterium infantis, Bifidobacterium longum, L. acidophilus, L. casei, Lactobacillus plantarum, Lactobacillus reuteri, L. rhamnosus, Lactobacillus salivarius, Lactobacillus sporogenes, and Streptococcus thermophilus was the best to reduce the incidence of diarrhea; multi-strain of Bacillus mesentericus, Clostridium butyricum, and Streptococcus faecalis for loss of appetite; multi-strain of B. longum, Lactobacillus bulgaricus, and S. thermophilus for constipation; multi-strain of Bifidobacterium bifidum, B. infantis, L. acidophilus, L. bulgaricus, L. casei, L. reuteri, and Streptococcus for taste disturbance; Saccharomyces boulardii for bloating; and multi-strain of Bifidobacterium breve, B. infantis, L. acidophilus, L. bulgaricus, L. casei, L. rhamnosus, and S. thermophilus for nausea/vomiting. CONCLUSIONS: Probiotics are recommended to supplement triple therapy in pediatrics, and the effectiveness of triple therapy is associated with specific probiotic supplementation.
Milk fermented by Lactobacillus species from Brazilian artisanal cheese protect germ-free-mice against Salmonella Typhimurium infection. L B Acurcio;S H C Sandes;R W Bastos;F M Sant'anna;S H S P Pedroso;D C Reis;Á C Nunes;G D Cassali;M R Souza;J R Nicoli. 2017. Benef Microbes. 8. PMID: 28789560

Ingestion of milks fermented by Lactobacillus strains showing probiotic properties is an important tool to maintain gastrointestinal health. In this study, Lactobacillus rhamnosus D1 and Lactobacillus plantarum B7, isolated from Brazilian artisanal cheese, were used as starters for the functional fermented milks to assess their probiotic properties in a gnotobiotic animal model. Male germ-free Swiss mice received a single oral dose of milk fermented by each sample, and were challenged with Salmonella Typhimurium five days afterwards. Milk fermented by both Lactobacillus strains maintained counts above 108 cfu/ml during cold storage. Lactobacillus strains colonised the gut of the germ-free-mice, maintaining their antagonistic effect. This colonisation led to a protective effect against Salmonella challenge, as demonstrated by reduced pathogen translocation and histological lesions, when compared to control group, especially for Lactobacillus rhamnosus D1. Additionally, mRNA expression of inflammatory (interferon gamma, interleukin (IL)-6, tumour necrosis factor alpha) and anti-inflammatory (transforming growth factor β1) cytokines was augmented in animals previously colonised and then challenged, when compared to other experimental groups. Lactobacillus plantarum B7 colonisation also promoted higher expression of IL-17, showing a proper maturation of colonised germ-free-mice immune system. IL-5 was stimulated by both strains' colonisation and not by S. Typhimurium challenge.
Dietary Fibers and Protective Lactobacilli Drive Burrata Cheese Microbiome. Fabio Minervini;Amalia Conte;Matteo Alessandro Del Nobile;Marco Gobbetti;Maria De Angelis. 2017. Appl Environ Microbiol. 83. PMID: 28842539

This study was aimed at improving the functional attributes and shelf life of burrata cheese by using protective lactobacilli (Lactobacillus plantarum LPAL and Lactobacillus rhamnosus LRB), fructooligosaccharides, and inulin. Six burrata cheeses were made using (i) the traditional protocol (control), (ii) the addition of 0.5% fructooligosaccharides and inulin (DF cheese), (iii) protective lactobacilli in milk alone (PL cheese), (iv) protective lactobacilli in milk and governing liquid (2PL cheese), (v) protective lactobacilli in milk and dietary fibers (DF_PL cheese), and (vi) protective lactobacilli in milk and governing liquid and dietary fibers (DF_2PL cheese). As expected, DF, DF_PL, and DF_2PL cheeses showed 1.5% of total fibers. Burrata cheeses produced by adding protective lactobacilli only in milk (PL and DF_PL cheeses) showed the lowest acidification during cheese making and storage. Lactic and acetic acids and ethanol were found at the lowest concentrations in these samples. Analyses of cultivable microbiota and the microbiome showed that protective lactobacilli reduced the house microbiota components (e.g., Streptococcus thermophilus, Lactococcus lactis, and Leuconostoc lactis) during cheese making and storage. Protective lactobacilli slowed the growth of staphylococci, coliforms, and Pseudomonas spp., especially in early storage. According to the different microbiome assemblies, burrata samples differed in peptide profiles and the levels of free amino acids. As shown by a sensory analysis, the addition of protective lactobacilli in milk improved the flavor and increased the shelf life of burrata cheese. In comparison to cheeses made using protective cultures only in milk, the shelf lives of those containing cultures also in the governing liquid were not further prolonged and they received lower acceptability scores by the panelists.IMPORTANCE This study provides more in-depth knowledge of the microbiome of burrata cheese and the set-up for a novel biotechnology using prebiotic dietary fibers and protective probiotic Lactobacillus plantarum LPAL and Lactobacillus rhamnosus LRB in milk. The biotechnology proposed in this study should be considered a useful tool to improve the functional value of burrata cheese. The use of protective lactobacilli in milk enhanced the flavor formation and shelf life of burrata cheese.
Effect of synbiotic supplementation on children with atopic dermatitis: an observational prospective study. M Dolores Ibáñez;Pablo Rodríguez Del Río;Diego González-Segura Alsina;Vicenç Villegas Iglesias. 2018. Eur J Pediatr. 177. PMID: 30259127

The objective of this observational single-cohort prospective study was to assess the effect of synbiotic supplementation for 8 weeks in children with atopic dermatitis (AD). The synbiotic product contained Lactobacillus casei, Bifidobacterium lactis, Lactobacillus rhamnosus, Lactobacillus plantarum, fructooligosaccharide, galactooligosaccharide, and biotin. Patients were examined at baseline and at 8 weeks. Effectiveness of treatment was assessed with the Scoring Atopic Dermatitis (SCORAD) index. A total of 320 children (mean age 5.1 years, range 0-12 years) were included. The mean (SD) SCORAD index decreased from 45.5 (15.5) at baseline to 19.4 (14.6) at the end of treatment (P < 0.001), VAS score for pruritus decreased from 5.7 (2.2) to 2.3 (2.2) (P < 0.001), and VAS score for sleep decreased from 3.1 (2.5) to 1.1 (1.8) (P < 0.001). Percentage of children with moderate-severe disease decreased from 92.4% at baseline to 28.1% at week 8. In the multiple linear regression analysis, higher baseline SCORAD index (OR 0.51; 95% CI 0.41-0.61) and higher adherence (OR 7.29; 95% CI 1.85-12.73) were significantly associated with greater decrease in SCORAD index.Conclusion: Supplementation with the multistrain synbiotic product may improve AD in children. What is known: • Pediatric atopic dermatitis (AD) is a common, troublesome condition with limited treatment options, which has been shown to be associated with dysbiosis in the intestinal microflora. • Results of controlled clinical trials (RCTs) on the effect of probiotics in children with AD have been disparate, although overall, the data favor probiotics over placebo, with multistrain supplements associated with better improvements in AD. What is new: • The results of this observational, prospective, open-label, single-cohort study on 320 children with AD younger than 12 years old suggest that supplementation with multistrain synbiotics (Lactobacillus casei, Bifidobacterium lactis, Lactobacillus rhamnosus, Lactobacillus plantarum, fructooligosaccharide, galactooligosaccharide, and biotin) helps to improve AD symptoms in children. • More than 80% of children experienced improvement in AD symptoms, as measured by Severity Scoring of Atopic Dermatitis (SCORAD) index and assessed by parents and physicians. The main predictive factors for improvement was adherence to synbiotic treatment and high baseline SCORE index; the change in SCORAD did not depend on age, gender, presence of concomitant treatment, duration, and type of AD (persistent vs with flares), other concomitant allergies or history of parental allergy.
Lactobacillus plantarum ZS2058 and Lactobacillus rhamnosus GG Use Different Mechanisms to Prevent Salmonella Infection in vivo. Junsheng Liu;Zhennan Gu;Fanfen Song;Hao Zhang;Jianxin Zhao;Wei Chen. 2019. Front Microbiol. 10. PMID: 30842764

Pathogen-induced infectious diseases pose great threats to public health. Accordingly, many studies have investigated effective strategies targeting pathogenic infections. We previously reported the preventive effects of Lactobacillus plantarum ZS2058 (ZS2058) and L. rhamnosus GG (LGG) against Salmonella spp. in a murine model. Here, we compared the mechanisms underlying the preventive effects of these Lactobacillus strains in vivo. Notably, reduced C-reactive protein levels were observed with both ZS2058 and LGG, which suggests abrogated anti-infection and inflammatory responses. ZS2058 more efficiently reduced the pathogenicity of Salmonella by increasing the level of propionic acid in feces and production of mucin 2 in the mouse colon and activity through the interleukin (IL)-23/IL-22 and IL-23/IL-17 pathways. Meanwhile, LGG more strongly alleviated gut inflammation, as indicated by changes in the levels of tissue necrosis factor (TNF)-α, IL-10 and myeloperoxidase (MPO) in infected mice. Moreover, both ZS2058 and LGG restored the levels of interferon (INF)-γ, a cytokine suppressed by Salmonella, albeit through different pathways. Our results demonstrate that ZS2058 and LGG prevent Salmonella infection via different mechanisms.
Effect of cold and frozen temperatures on artisanal goat cheese containing probiotic lactic acid bacteria isolates (Lactobacillus plantarum TW14 and Lactobacillus rhamnosus TW2). Triana Setyawardani;Juni Sumarmono;Kusuma Widayaka. 2019. Vet World. 12. PMID: 31089311

Aim: The research was conducted to determine the effect of temperature and storage duration on the physicochemical, lipolytic, microbiological, and proteolytic characteristics of goat cheese made using Lactobacillus plantarum TW14 and Lactobacillus rhamnosus TW2 bacteria. Materials and Methods: The cheese was stored at 4°C and -20°C for 0, 15, 30, 45, and 60 days. Observations were made on its physicochemical, lipolysis, and microbiological characteristics. The proteolysis pattern was measured with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Results: The protein, fat, ash and total solids levels of cold-stored cheese were higher than the frozen-stored one. The frozen-stored cheese's free fatty acids (FFA) and acid degree value (ADV) levels are lower than those of the cold-stored cheese as indicated by the partial lipolysis event. The total yeast in the frozen-stored cheese is lower than that in the frozen-stored cheese. Finally, the electrophoresis profile indicates that proteolysis of the frozen-stored cheese is formed since there have been detected αs1-casein, αs2-casein, β-casein, and κ-casein in the casein breakdown during the 60-day storage. Conclusion: The physicochemical characteristics of cold-stored cheese are better than the cheese stored at frozen temperature. However, frozen-stored cheese produces lower FFA and ADV than cold-stored cheese and lipolysis occurs only partially.
Sugar fermentation in probiotic bacteria--an in vitro study. M Hedberg;P Hasslöf;I Sjöström;S Twetman;C Stecksén-Blicks. 2008. Oral Microbiol Immunol. 23. PMID: 18954354

INTRODUCTION: Food supplemented with probiotic bacteria is a rapidly growing sector of the market. The aim of the present study was to evaluate and compare the acid production of selected probiotic strains available in commercial products. METHODS: Six Lactobacillus strains (Lactobacillus plantarum 299v and 931; Lactobacillus rhamnosus GG and LB21; Lactobacillus paracasei subsp. paracasei F19, and Lactobacillus reuteri PTA 5289) were cultivated at 37 degrees C in an anaerobic atmosphere on Man, Rogosa, Shape (MRS) agar for 48 h or MRS broth for 16 h. After centrifugation, the cells were washed and resuspended in sterile phosphate-buffered saline and immediately subjected to a fermentation assay with 12 different carbohydrates (nine sugars and three sugar alcohols) in microtiter plates with a pH indicator. The plates were examined for color changes after 24, 48, and 72 h of incubation under aerobic and anaerobic conditions. Three scores were used: negative (pH > 6.8); weak (pH 5.2-6.8), and positive (pH < 5.2). The strains were characterized with the API 50 CH system to confirm their identity. RESULTS: L. plantarum fermented all the sugars except for melibiose, raffinose, and xylitol. Both L. rhamnosus strains were generally less active although L. rhamnosus GG was slightly more active than strain LB21 in the 5% CO(2) setting. The latter strain exhibited negative reactions for sucrose, maltose, arabinose, and sorbitol under anaerobic conditions. The assays with L. paracasei and L. reuteri had negative or weak reactions for all tested sugars under both aerobic and anaerobic conditions. CONCLUSION: The metabolic capacity to form acid from dietary sugars differed significantly between the various probiotic strains.