Solution is here!
What is Lactospore® ?
• LactoSpore is a shelf-stable, clinically validated commercial probiotic preparation from Sabinsa Inc., containing Lactic Acid producing microbial preparation from Bacillus coagulans MTCC 5856 (earlier known as Lactobacillus sporogenes) which is known for its ability to form extremely resistant spores.
• B. coagulans is a spore-forming, Gram-positive, facultative anaerobic, L-(+)-lactic acid producing organism.
• Products supplemented with B. coagulans can be stored at room temperature without any deleterious effect on their viability.
• It is one of the most promising spore-forming probiotics, currently available as a dietary supplement worldwide (FDA, 2015) and has been reported to support healthy digestive and immune functions.
• Spores confers higher resistance to technological stresses encountered during industrial production and storage processes and greater protection against hostile gastric and intestinal conditions (pH, bile and digestive enzymes).
How does Lactospore work?
• Probiotics are known to produce the short chain fatty acids, an action that results in decreased luminal pH and production of bactericidal proteins. Butyric acid, a by-product of bacterial fermentation of fibre, has been shown to nourish colonic enterocytes, enhancing mucosal integrity. Majeed et al. also reported that B. coagulans MTCC 5856 produced short chain fatty acids (acetate, butyrate, and propionate) by fermenting plant based fibres. Despite availability of ample data, the precise mechanism of action by the probiotic in IBS and other gastrointestinal disorders still remains to be confirmed.
• Subsequent to oral administration, B. Coagulans MTCC 5856 arrives in the stomach in its spore form, where it is exposed to the stomach’s churning action and acidic pH that causes the spore coating to absorb water, swell and begin the germination process. Upon arrival in the duodenum, the spores germinate and multiply rapidly after germination.
• B. coagulans MTCC 5856 was found to decrease clinical symptoms like bloating, vomiting, diarrhoea, abdominal pain and stool frequency in a multi-centre randomised clinical trial which suggested its safe and effective use in the management of diarrhoea, predominant irritable bowel syndrome (IBS) (Majeed et al., 2014).
• Similar to Lactobacillus, B. coagulans has been shown to display immuno-regulatory effects that could potentially impact the health of the skin. The incubation of peripheral blood mononuclear cells (PBMCs) and polymorphonuclear (PMN) cells with the supernatant and cell wall fragments of B. coagulans promoted mature phenotypes of antigen-presenting cells and inhibited spontaneous and stress-induced reactive oxygen species (ROS) formation. We are well aware that ROS and oxidative stress play a role in acne, making this an intriguing finding with potential benefit for the acne patient. [15]
• It is a well-known fact that Bacillus species has the ability to form spores when growth conditions are not in their favour and can remain in the dormant stage for many years. However, when the growth conditions are favourable such as specific nutrients, pH, temperature and moisture, it can cause spores to become vegetative and return to life through the process of germination. (Nicholson et al., 2000; Setlow, 2014).
• Despite availability of ample data, the precise mechanism of action by the probiotic in IBS and other gastrointestinal disorders still remains to be confirmed
Evaluation of the stability of Bacillus coagulans MTCC 5856 during processing and storage of functional foods
Dr. Muhammed Majeed et al [2016] International Journal of Food Science & Technology Materials & Methods
• Bacillus coagulans MTCC 5856 samples used in the study were manufactured by Sami Labs Limited (Bangalore, India) by following a proprietary, in-house developed, good manufacturing process. Pure B. coagulans MTCC 5856 spores were spray– dried and standardised with food grade maltodextrin (Sanwa Starch Co. Ltd. Kashihara, Nara, Japan) to achieve the desired concentration of 15 9 109 CFU g1 of powdered form for the finished product. This standardised preparation was used throughout the study until otherwise stated.
• After every time interval for the stability studies, 1.0 g of sample (baked food or other studied product containing B. coagulans MTCC 5856) was thoroughly mixed in sterile saline (0.9% NaCl, w/v) and then incubated in water bath for 30 min at 75 °C, followed by immediate cooling to below 45 °C. This suspension was further serially diluted in sterile saline and the viable count was enumerated by plating on glucose yeast extract agar by pour plate method. The plates were incubated at 37 °C for 48–72 h. Each analysis was performed in triplicate at two different occasions. Average mean of spore viable counts are expressed in log10 CFU.
• Waffles: Prepared batter was poured in the waffle iron and spread evenly and baked for 2 min or until the waffle was golden brown. Waffles were stored in freezer (20 °C ± 2) and the viable count of B. coagulans MTCC 5856 was enumerated by dissolving in sterile saline (0.9% NaCl, w/v). Samples were analysed at 0, 1, 2, 3, 6 and 12 months by serial dilution method.
• Muffin: It was baked for 20–25 min at 205 °C and then cooled completely and stored in the freezer (20 °C ±2). The viable count of B. coagulans MTCC 5856 was enumerated by dissolving muffins in sterile saline (0.9% NaCl, w/v) followed by serial dilution method. Samples were analysed at 0, 1, 2, 3, 6 and 12 months.
• Brewed coffee: Coffee powder containing B.coagulans MTCC 5856 was added to microbial free water (preheated at 90 2 °C) and temperature was maintained at 90 ± 2 °C for 2 min using boiling water bath. Samples were then cooled to 77 ± 2 °C and same temperature was maintained for 4h using boiling water bath. Samples were withdrawn before and after brewing followed by holding the temperature at 77 °C for 0.5,1, 2 and 4 h. Samples were analysed for viable spore counts by serial dilution method.
• Chocolate fudge frosting and hot fudge topping: Chocolate fudge frosting was heated at 60 °C temperature and then B. coagulans MTCC 5856 (0.071%, w/w) was added and uniformly mixed for 15 min. Similarly, hot fudge topping was heated at 60 °C, mixed with B. coagulans MTCC 5856 and mixed for 15 min. All samples were stored at room temperature and tested at 0,1,2,3,6 and 12 months for viable spore counts by serial dilution method.
Evaluation of the stability of Bacillus coagulans MTCC 5856 during processing and storage of functional foods
Peanut Butter and strawberry preserves: Peanut butter was heated at 30– 35 °C and B. coagulans MTCC 5856 (0.081%, w/w) was added followed by blending for 60 min. This was filled in airtight sterile jars and stored at room temperature. Similarly, strawberry preserve was preheated at 75 °C and B. coagulans MTCC 5856 (0.114%, w/w) was added to it and blended for 15 min and then filled in a sterile glass jar. All the samples were stored at room temperature and analysed at 0, 1, 2, 3, 6 and 12 months for viable spore counts by serial dilution method.
• Apple Juice: Apple juice containing 17% of concentrated apple juice with no added sugar, colour and preservatives was procured and tested for being free of any microbial contamination. The pH was measured to be 3.5. One gram of B. coagulans MTCC 5856 was added to 1L of apple juice and mixed well for 30 min. Aseptically 40mL sample was transferred to a sterile polypropylene tube and sealed. Samples were stored in refrigerated condition (4–6 °C) and tested at 0, 1, 2, 3 and 6 months of time interval for viable spore count by serial dilution method
• Concentrated glucose syrup: Concentrated glucose syrup was prepared by adding 85.0 g of glucose into 15.0 g of water and slowly heated to 70 °C for 10 min then boiled for 10 min. It was then cooled down to 45-50 °C and then B. coagulans MTCC 5856 was added and blended for 30 min. Aseptically, 50g of sample was transferred to sterile polypropylene bottles and sealed. These samples were stored at accelerated (40 ± 2 °C, RH = 75 ± 5%), room temperature (30 ± 2 °C, RH = 65 ± 5%) and refrigerated (4 ± 3°C). Samples were analysed at 0, 1, 2, 3, 6, 12, 18 and 24 months for viable spore counts using serial dilution method.
Results:
• The stability of B. coagulans MTCC 5856 was evaluated in waffles, hot fudge topping, chocolate fudge frosting and banana muffin mix during manufacturing process and storage at respective conditions.
• There was 91% survival of B. coagulans MTCC 5856 during the manufacturing of waffles and retained 86% viability up to 12 months of storage at frozen conditions (-20 °C).
• Hot fudge topping, chocolate fudge frosting and banana muffin mix retained >95% viability during their manufacturing and storage.
• There was a 13% reduction from the initial count of CFUs as observed after brewing at 90°C for 2 min. No significant reduction in the viability of B. coagulans MTCC 5856 after brewing till 1h at holding temperature.
• The viability of B. coagulans MTCC 5856 in peanut butter and strawberry preserves was retained over 97% upto 12 months of storage at room temperature
• Bacillus coagulans MTCC 5856 was formulated in pure canola oil and stored at room temperature. There was no significant reduction in the viability (more than 95% upto 12 months) of B. coagulans MTCC 5856 either during blending or storage.
• B. coagulans MTCC 5856 was found to be stable in apple juice and retained more than 99% viability upto 6 months of storage at refrigerated conditions (4– 6 °C)
Indications for Lactospore®
Lactospore® has been clinically proven effective for:
• Diarrhoea (Adult and Neonates)
• Antibiotic associated diarrhoea
• Constipation • Blood cholesterol management
• Irritable Bowel Syndrome (IBS)
• Immune support
• Blood glucose management
• Neonatal jaundice
• Non-specific vaginitis (Lactic Acid decreases pH of vagina, giving protective effect)