Inducia

INDUCIA®

The lactic acid bacterium Lactobacillus plantarum INDUCIA® can be used in foodstuff or in dietary supplements

Lactobacillus plantarum INDUCIA® (DSM21379)
 

Origin, identification, deposition, ownership

Lactobacillus plantarum INDUCIAâ is of healthy human origin. It has been discovered and investigated by researchers of BioCC LLC in collaboration with researchers from the University of Tartu.

Strain Lactobacillus plantarum INDUCIAâ was identified on the basis of biochemical activity with API 50CHL System (bioMérieux, France) kit as Lactobacillus plantarum (coincidence with the type strain: excellent, ID %-99.9, T index -0.81).

The molecular identification of the strain INDUCIAâ as Lactobacillus plantarum was confirmed by ITS-PCR (Internal-Transcribed Spacer Polymerase Chain Reaction) and 16S rRNA sequencing

L. plantarum INDUCIAâ is deposited in Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ; Braunschweig, Germany), under the number DSM 21379, in accordance with Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure, by the BioCC LLC on 16.04.2008.

L.plantarum INDUCIAâ is the property and trademark of the BioCC LLC.

Results of clinical trials

Mechanisms of action

Lactobacillus plantarum strain INDUCIA ® is a multipotent microbe of a healthy child origin, the most prominent feature of which is the ability to reduce blood cholesterol level and with strong antioxidative and antimicrobial activity.

Excessive levels of LDL cholesterol lead to the formation of atherosclerotic plaques ultimately followed by myocardial infarction or stroke.

Oxidized LDL cholesterol (oxLDL) is a marker of oxidative stress specific to LDL particles. Elevated Ox-LDL level is associated with development of cardiovascular diseases.

The reduction of LDL and ox-LDL content of cholesterol are important for prevention of atherosclerosis and maintenance of cardiovascular health.

  • INDUCIA ® helps to maintain the cardiovascular system by reducing the level of LDL cholesterol in the blood via hydrolyzing bile salts due to BSH (bile salt hydrolase) activity and by assimilation of dietary cholesterol in the intestine.
  • INDUCIA® reduces the level of oxidised low-density lipoprotein (ox-LDL) and the oxidative stress index of the human body
  • INDUCIA® supports the antioxidative network of the human body due to its strong antioxidative properties and the presence of a complete intracellular glutathione system.

Reactive oxygen species (ROS) are products of normal cellular metabolism and derive from various sources in different cellular compartments. Oxidative stress resulting from an imbalance between ROS generation and antioxidant defence mechanisms is important in the pathogenesis of cardiovascular diseases. The important components of the cellular antioxidative defense system are reduced glutathione and antioxidative enzymes like glutathione peroxidase.

Additionally, consumption of Lactobacillus plantarum INDUCIA®

  • helps to maintain and restore the balance of intestinal microflora.
  • increases the amount of useful lactobacilli in the gut, which ensures the breakdown of more absorbable compounds of nutrients.
  • enhances the human body’s natural resistance to several enteropathogens due to strong antimicrobial activity through production of a variety of antimicrobial compounds (organic acids and hydrogen peroxide, plantaricins)
  • In food products plantarum INDUCIA® reduces the risk of foodborne infections by suppressing potentially pathogenic bacteria due to antimicrobial activity against enteric pathogens like listeria, salmonella and shigella and psychrophilic microbes.
  • INDUCIA® is able to prevent the germination of the spores of Clostridium difficile and the proliferation of vegetative cells.

Clostridium difficile is an anaerobic spore-forming intestinal pathogen. C. difficile infection is one of the most prevalent healthcare-associated infections in hospitals and nursing homes. In order to cause disease, C. difficile spores must germinate and return to vegetative cell growth and secrete toxins. Twenty-five percent of patients infected with C. difficile develop serious health conditions such as pseudomembraneous colitis. Clostridium difficile-associated diarrhoea increases mortality rates, lengthens hospitalization and dramatically increases overall health care costs.

Biosafety and tolerability

Lactobacilli incl. the species Lactobacillus plantarum have been consumed for centuries in the form of fermented foods and are generally recognised as safe (FAO/WHO, 2002). The species L. plantarum is considered by the EFSA to be suitable for the Qualified Presumption of Safety (QPS) approach (EFSA, 2007 and 2012).

At the same time, Lactobacillus plantarum as a species is a normal component of the intestinal microbiota in humans. The human (healthy child) origin of the strain Lactobacillus plantarum INDUCIA ® is a prerequisite for its harmless status and suitability for oral application.

The science-based safety testing of L. plantarum INDUCIA ® has been carried out in test tubes and on laboratory animals.

The antibiotic susceptibility profile of L. plantarum INDUCIA ® was determined according to ISO 10932:2010 with VetMIC Lact-1 and VetMIC Lact-2 plates (SVA National Veterinary Institute, Uppsala, Sweden). The minimum inhibitory concentrations for L. plantarum INDUCIAâ did not exceed the MIC cut-off values proposed for the species Lactobacillus plantarum proposed by the EFSA (EFSA 2012).

The biosafety of L. plantarum INDUCIA ® has been tested in a mouse toxicity model of 30 days of feeding. No translocation of bacteria into blood and no pathological shifts were found by the morphological and histological evaluation of the liver, spleen and gut.

The safety and tolerability of L. plantarum INDUCIA ® has been proved in two age groups of a general healthy adult population (age 20…60 y). In human intervention trials, no discomfort, abdominal pain or other negative symptoms, nor increased inflammation markers (hs-CRP, WBC count; IgG, IgM and IgE levels) were reported. Consumption of Lactobacillus plantarum INDUCIA ® had no negative impact on kidney and liver function nor affected according parameters (ASAT, ALAT albumine, blood serum creatinine) over the course of 3 weeks and 8 weeks of daily consumption of L. plantarum INDUCIA® in a dose of 2 to 5 x 109 viable microbial cells.

The strain L. plantarum INDUCIA® was elaborated as a probiotic by BioCC LLC in cooperation with the University of Tartu Departments of Microbiology and Biochemistry according to FAO /WHO (2002) guidelines.

Faecal recovery/survival in GIT

One of the selection criterions for the probiotics is the ability to survive in the GI tract in order to be effective in therapeutic actions and carry on normal metabolic activity after consumption. The high faecal recovery of a probiotic strain after consumption reflects the viability and good survival of a particular strain in the GI tract.

L. plantarum INDUCIA® is able to survive the human gastrointestinal passage. RT–PCR analysis with strain-specific primers confirmed the appearance of L. plantarum INDUCIA® in the faeces of 40% of adult volunteers after 4 weeks and in 52% after 8 weeks of daily consumption of L. plantarum INDUCIA® with functional food in a daily dose of at least 2 x 109 CFU.

Results of clinical trials

Dyslipidemia remains one of the most firmly established risk factors for coronary heart disease, stroke and diabetes. Excessive levels of LDL cholesterol lead to the formation of atherosclerotic plaques ultimately followed by myocardial infarction or stroke. Oxidized LDL cholesterol (oxLDL) is a marker of oxidative stress specific to LDL particles.

Oxidative stress occurs when abnormally high levels of reactive oxygen species are generated, resulting in DNA, protein and lipid damage. Excessive oxidative stress is important in the pathophysiology of a variety of human diseases incl. cardiovascular diseases.

The health effects of L. plantarum INDUCIA® have been tested on over 500 people.

According to the human intervention trials (ISRCTN79645828 and ISRCTN26344255) the composition comprising L. plantarum INDUCIA® has a cardio-protective effect through reduction of LDL cholesterol in blood and simultaneously protects the human body from oxidative damage and enhances anti-oxidative activity by reducing the oxidative stress markers oxLDL, OSI and TPX.

 

Antagonistic activity of L. plantarum INDUCIA® against C difficile

In vitro and animal studies

Antagonistic activity of L. plantarum INDUCIA® against vegetative cells of C difficile.  

Clostridium difficile infection (CDI) is one of the most prevalent healthcare-associated infections in hospitals and nursing homes.

We tested a) the susceptibility of C. difficile strains of different origin and the intestinal probiotic Lactobacillus plantarum INDUCIAâ (DSM 21379) on various antimicrobial preparations incl. metronidazole and vancomycin; b) the susceptibility of C. difficile strains on the antagonistic effects of the probiotic L. plantarum INDUCIAâ, prebiotic xylitol (Xyl) and their combination as a synbiotic (Syn) product; c) the suppression of germination of C. difficile spores in vitro and in vivo in an animal model of C. difficile infection with INDUCIAâ, Xyl and Syn treatment.

The clinical isolates (n = 12) of C. difficile, VPI strain 10463 (ATCC 43255) and epidemic strain (M 13042) were susceptible and contrarily L. plantarum INDUCIAâ resistant to vancomycin, metronidazole and ciprofloxacin.

The intact cells of INDUCIAâ, natural and neutralized cell free supernatant inhibited in vitro the growth of tested C. difficile reference strain VPI and clinical isolates of C. difficile after co-cultivation. Incubation of INDUCIAâ in the media with 5% Xyl fully stopped germination of spores of C. difficile VPI strain after 48 hours.

In an infection model the 48 hamsters were administered ampicillin (30 mg/kg) and 10-30 spores of C. difficile VPI strain. They also received five days before and after the challenge a pretreatment with a synbiotic (single daily dose of L. plantarum INDUCIAâ 1 ml of 1010 CFU/ml and 20% xylitol in 1 ml by orogastric gavage). The survival rate of hamsters was increased to 78% compared to 13% (p = 0.003) survival rate of specimens who received no treatment. When administered Xyl the survival rate of hamsters reached 56% vs.13% (p = 0.06). In both Syn (6/9, p = 0.003) and Xyl (3/9, p = 0.042) groups the number of specimens not colonized with C. difficile significantly increased.

In conclusion, the combination of xylitol with L. plantarum INDUCIAâ suppresses the germination of spores and outgrowth into vegetative toxin-producing cells of C. difficile and reduces the colonization of the gut with the pathogen.

For details see: Rätsep et al., 2017 A combination of the probiotic and prebiotic product can prevent the germination of Clostridium difficile spores and infection. Anaerobe 47: 94-103

 

Technological properties

L. plantarum INDUCIAâ tolerates technological processing well and copes well with different stressors. The strain withstands the manufacturing process of food, is able to multiply at relatively low (+12oC) and high (+45oC) temperatures and in a carbohydrate-scarce medium. This property makes the strain usable as a probiotic adjunct starter in functional food.

BioCC has developed concepts for the creation INDUCIAâcontaining food product(s) and dietary supplement(s).

Patents

Estonian patent EE05341

Granted: 16.08.2010

Russian patent RU2486234

Granted: 27.06.2013

Korean patent KR10-1595042

Granted: 14.01.2016

European patent EP2288360

Granted> 14.12.2011

Selected publications

Rätsep , S. Kõljalg, E. Sepp, I. Smidt, K. Truusalu, E. Songisepp, J. Stsepetova, P. Naaber, R.H. Mikelsaar, M. Mikelsaar. A 

combination of the probiotic and prebiotic product can prevent the germination of Clostridium difficile spores and infection. Anaerobe 47 (2017) 94-103

Kullissar, T; Songisepp, E; Zilmer, M. (2012). 

Probiotics and oxidative stress. In: Oxidative stress -environmental induction and dietary antioxidants. (203−222). InTech – Open Access Publisher

Rätsep, M.; Hütt, P.; Avi, R.; Utt, M.; Songisepp, E. (2011). 

Antimicrobial properties ofLactobacillus plantarum Tensia (DSM 21380) and Inducia (DSM 21379). In: Mendez-Vilas, A. (Ed.). Science and Technology against Microbial Pathogens. Research, Development and Evaluation. (393−397). World Scientific Publ Co Pte Ltd.

Songisepp, E.; Mikelsaar, M.; Zilmer, M.; Hütt, P.; Smidt, I.; Stsepetova, J.; Truusalu, K.; Rätsep, M.; Kilk, K.; Utt, M.; Üksti, J.; Kõljalg, S., Zilmer, K. (2008). 

Lactobacillus plantarum TENSIA (DSM 21380) and plantarum INDUCIA (DSM 21379) as novel probiotics. FIL-IDF World Dairy Summit, Mecixo City, Mexico, 11-14 November 2008. Mexico City