Selective Breeding of Oxygen-Tolerant and Oxalate-Degrading Lactic Acid Bacteria by Protoplast Fusion

Biﬁdobacterium lactiswith oxalate-degrading capacity can efficiently reduce the oxalate in vivo, and it can be used to prevent and treat kidney stone diseases. WhileBiﬁdobacterium lactisis poorly oxygen-tolerant, which hinders it from being as microbial ecological agents. To obtain oxygen-tolerant and oxalate-degrading lactic acid bacteria, protoplast fusion technology was used betweenB. lactisandL. acidophilus.Under the optimum conditions of protoplast fusion with PEG 6000 concentration 50%, the fusion time 7 min, the fusion temperature 30°C, the concentration of CaCl20. 02mol/ L and the concentration of MgCl20.5mol/ L, the fusion rate reached 7.6%, and three oxygen-tolerant fusant showing that the level of oxalate degradation were similar withB. lactiswas obtained. The fusants of SZY1-7 and SZY2-1 could tolerance to pH 2.5 and 0.5% (w/v) bile salt.

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Isolation of lactic acid bacteria from swine milk and characterization of potential probiotic strains with antagonistic effects against swine-associated gastrointestinal pathogens

Canadian Journal of Microbiology ◽

10.1139/cjm-2015-0811 ◽

2016 ◽

Vol 62 (6) ◽

pp. 514-524 ◽

Cited By ~ 8

Author(s):

Sandra Rayén Quilodrán-Vega ◽

Julio Villena ◽

José Valdebenito ◽

María José Salas ◽

Cristian Parra ◽

...

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Human Milk ◽

Salmonella Enterica ◽

Enterohemorrhagic Escherichia Coli ◽

Salmonella Infection ◽

Antagonistic Effects ◽

Gastrointestinal Pathogens

Probiotics are usually isolated from the gastrointestinal tract of humans and animals. The search of probiotics in human milk is a recent field of research, as the existence of the human milk microbiome was discovered only about a decade ago. To our knowledge, no reports regarding the potential probiotic effect of bacteria from swine milk have been published. In this work, we isolated several lactic acid bacteria from swine milk and evaluated them for them potential as probiotics. Among the isolated strains, Lactobacillus curvatus TUCO-5E showed antagonistic effects against swine-associated gastrointestinal pathogens. TUCO-5E was able to reduce the growth of enterotoxigenic and enterohemorrhagic Escherichia coli strains as well as pathogenic salmonella. In vitro exclusion and displacement assays in intestinal epithelial cells showed a remarkable antagonistic effect for L. curvatus TUCO-5E against Salmonella sp. strain TUCO-I7 and Salmonella enterica ATCC 13096. Moreover, by using a mouse model of Salmonella infection, we were able to demonstrate that preventative administration of L. curvatus TUCO-5E for 5 consecutive days was capable of decreasing the number of Salmonella enterica serovar Typhimurium in the liver and spleen of treated mice, compared with the controls, and prevented dissemination of the pathogen to the blood stream. Therefore, we have demonstrated here that swine milk is an interesting source of beneficial bacteria. In addition, the results of this work suggest that L. curvatus TUCO-5E is a good candidate to study in vivo the protective effect of probiotics against intestinal infection and damage induced by Salmonella infection in the porcine host.

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In vivo and In vitro Mitigation Effects of Lactic Acid Bacteria Derived from Fresh Vegetables on Aflatoxins

Food Hygiene and Safety Science (Shokuhin Eiseigaku Zasshi) ◽

10.3358/shokueishi.62.148 ◽

2021 ◽

Vol 62 (5) ◽

pp. 148-156

Author(s):

Kenji Oonaka ◽

Naoki Kobayashi ◽

Yosuke Uchiyama ◽

Mioko Honda ◽

Shiro Miyake ◽

...

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Fresh Vegetables

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Capacity of Human Nisin- and Pediocin-Producing Lactic Acid Bacteria To Reduce Intestinal Colonization by Vancomycin-Resistant Enterococci

Applied and Environmental Microbiology ◽

10.1128/aem.02150-07 ◽

2008 ◽

Vol 74 (7) ◽

pp. 1997-2003 ◽

Cited By ~ 85

Author(s):

Mathieu Millette ◽

Gilbert Cornut ◽

Claude Dupont ◽

François Shareck ◽

Denis Archambault ◽

...

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

In Vivo Study ◽

Control Group ◽

Intestinal Colonization ◽

Strong Activity ◽

Vancomycin Resistant Enterococci ◽

Significant Difference ◽

Vancomycin Resistant

ABSTRACT This study demonstrated the capacity of bacteriocin-producing lactic acid bacteria (LAB) to reduce intestinal colonization by vancomycin-resistant enterococci (VRE) in a mouse model. Lactococcus lactis MM19 and Pediococcus acidilactici MM33 are bacteriocin producers isolated from human feces. The bacteriocin secreted by P. acidilactici is identical to pediocin PA-1/AcH, while PCR analysis demonstrated that L. lactis harbors the nisin Z gene. LAB were acid and bile tolerant when assayed under simulated gastrointestinal conditions. A well diffusion assay using supernatants from LAB demonstrated strong activity against a clinical isolate of VRE. A first in vivo study was done using C57BL/6 mice that received daily intragastric doses of L. lactis MM19, P. acidilactici MM33, P. acidilactici MM33A (a pediocin mutant that had lost its ability to produce pediocin), or phosphate-buffered saline (PBS) for 18 days. This study showed that L. lactis and P. acidilactici MM33A increased the concentrations of total LAB and anaerobes while P. acidilactici MM33 decreased the Enterobacteriaceae populations. A second in vivo study was done using VRE-colonized mice that received the same inocula as those in the previous study for 16 days. In L. lactis-fed mice, fecal VRE levels 1.73 and 2.50 log10 CFU/g lower than those in the PBS group were observed at 1 and 3 days postinfection. In the P. acidilactici MM33-fed mice, no reduction was observed at 1 day postinfection but a reduction of 1.85 log10 CFU/g was measured at 3 days postinfection. Levels of VRE in both groups of mice treated with bacteriocin-producing LAB were undetectable at 6 days postinfection. No significant difference in mice fed the pediocin-negative strain compared to the control group was observed. This is the first demonstration that human L. lactis and P. acidilactici nisin- and pediocin-producing strains can reduce VRE intestinal colonization.

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In vitro and in vivo evaluation of lactic acid bacteria of aquatic origin as probiotics for turbot (Scophthalmus maximus L.) farming

Fish & Shellfish Immunology ◽

10.1016/j.fsi.2014.10.007 ◽

2014 ◽

Vol 41 (2) ◽

pp. 570-580 ◽

Cited By ~ 39

Author(s):

Estefanía Muñoz-Atienza ◽

Carlos Araújo ◽

Susana Magadán ◽

Pablo E. Hernández ◽

Carmen Herranz ◽

...

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Scophthalmus Maximus ◽

In Vivo Evaluation ◽

Turbot Scophthalmus Maximus

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Effect of Lactic Acid Bacteria on the Pharmacokinetics and Metabolism of Ginsenosides in Mice

Pharmaceutics ◽

10.3390/pharmaceutics13091496 ◽

2021 ◽

Vol 13 (9) ◽

pp. 1496

Author(s):

Ji-Hyeon Jeon ◽

Jaehyeok Lee ◽

Jin-Hyang Park ◽

Chul-Haeng Lee ◽

Min-Koo Choi ◽

...

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Formation Rate ◽

Plasma Concentrations ◽

Control Level ◽

In Vitro Fermentation ◽

Fecal Recovery ◽

Fermentation Test

This study aims to investigate the effect of lactic acid bacteria (LAB) on in vitro and in vivo metabolism and the pharmacokinetics of ginsenosides in mice. When the in vitro fermentation test of RGE with LAB was carried out, protopanaxadiol (PPD) and protopanaxadiol (PPD), which are final metabolites of ginsenosides but not contained in RGE, were greatly increased. Compound K (CK), ginsenoside Rh1 (GRh1), and GRg3 also increased by about 30%. Other ginsenosides with a sugar number of more than 2 showed a gradual decrease by fermentation with LAB for 7 days, suggesting the involvement of LAB in the deglycosylation of ginsenosides. Incubation of single ginsenoside with LAB produced GRg3, CK, and PPD with the highest formation rate and GRd, GRh2, and GF with the lower rate among PPD-type ginsenosides. Among PPT-type ginsenosides, GRh1 and PPT had the highest formation rate. The amoxicillin pretreatment (20 mg/kg/day, twice a day for 3 days) resulted in a significant decrease in the fecal recovery of CK, PPD, and PPT through the blockade of deglycosylation of ginsenosides after single oral administrations of RGE (2 g/kg) in mice. The plasma concentrations of CK, PPD, and PPT were not detectable without change in GRb1, GRb2, and GRc in this group. LAB supplementation (1 billion CFU/2 g/kg/day for 1 week) after the amoxicillin treatment in mice restored the ginsenoside metabolism and the plasma concentrations of ginsenosides to the control level. In conclusion, the alterations in the gut microbiota environment could change the ginsenoside metabolism and plasma concentrations of ginsenosides. Therefore, the supplementation of LAB with oral administrations of RGE would help increase plasma concentrations of deglycosylated ginsenosides such as CK, PPD, and PPT.

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A SH3_5 Cell Anchoring Domain for Non-recombinant Surface Display on Lactic Acid Bacteria

Frontiers in Bioengineering and Biotechnology ◽

10.3389/fbioe.2020.614498 ◽

2021 ◽

Vol 8 ◽

Author(s):

Pei Kun Richie Tay ◽

Pei Yu Lim ◽

Dave Siak-Wei Ow

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Surface Display ◽

Bioactive Molecules ◽

Gastric Digestion ◽

Functional Protein ◽

Fluorescent Reporter ◽

Surface Conditions ◽

Bacterial Surface

Lactic acid bacteria (LAB) are a group of gut commensals increasingly recognized for their potential to deliver bioactive molecules in vivo. The delivery of therapeutic proteins, in particular, can be achieved by anchoring them to the bacterial surface, and various anchoring domains have been described for this application. Here, we investigated a new cell anchoring domain (CAD4a) isolated from a Lactobacillus protein, containing repeats of a SH3_5 motif that binds non-covalently to peptidoglycan in the LAB cell wall. Using a fluorescent reporter, we showed that C-terminal CAD4a bound Lactobacillus fermentum selectively out of a panel of LAB strains, and cell anchoring was uniform across the cell surface. Conditions affecting CAD4a anchoring were studied, including temperature, pH, salt concentration, and bacterial growth phase. Quantitative analysis showed that CAD4a allowed display of 105 molecules of monomeric protein per cell. We demonstrated the surface display of a functional protein with superoxide dismutase (SOD), an antioxidant enzyme potentially useful for treating gut inflammation. SOD displayed on cells could be protected from gastric digestion using a polymer matrix. Taken together, our results show the feasibility of using CAD4a as a novel cell anchor for protein surface display on LAB.

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Oral Administration of Lactococcus lactis Producing Interferon Type II, Enhances the Immune Response Against Bacterial Pathogens in Rainbow Trout

Frontiers in Immunology ◽

10.3389/fimmu.2021.696803 ◽

2021 ◽

Vol 12 ◽

Author(s):

Alvaro Santibañez ◽

Diego Paine ◽

Mick Parra ◽

Carlos Muñoz ◽

Natalia Valdes ◽

...

Keyword(s):

Immune Response ◽

Lactic Acid ◽

Rainbow Trout ◽

Lactic Acid Bacteria ◽

Atlantic Salmon ◽

Oral Administration ◽

Bacterial Infection ◽

Type Ii ◽

Recombinant Interferon

Lactic acid bacteria are a powerful vehicle for releasing of cytokines and immunostimulant peptides at the gastrointestinal level after oral administration. However, its therapeutic application against pathogens that affect rainbow trout and Atlantic salmon has been little explored. Type II interferon in Atlantic salmon activates the antiviral response, protecting against viral infection, but its role against bacterial infection has not been tested in vivo. In this work, through the design of a recombinant lactic acid bacterium capable of producing Interferon gamma from Atlantic salmon, we explore its role against bacterial infection and the ability to stimulate systemic immune response after oral administration of the recombinant probiotic. Recombinant interferon was active in vitro, mainly stimulating IL-6 expression in SHK-1 cells. In vivo, oral administration of the recombinant probiotic produced an increase in IL-6, IFNγ and IL-12 in the spleen and kidney, in addition to stimulating the activity of lysozyme in serum. The challenge trials indicated that the administration of the IFNγ-producing probiotic doubled the survival in fish infected with F. psychrophilum. In conclusion, our results showed that the oral administration of lactic acid bacteria producing IFNγ managed to stimulate the immune response at a systemic level, conferring protection against pathogens, showing a biotechnological potential for its application in aquaculture.

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