You are on page 1of 84

SPORLAC

I n d i a ' s F i r s t P r o b i o t i c

C e l e b r a t i n g

4 0

Y e a r s

1949, Just after World War II... JAPAN

rof. Mashita and Prof. Nakayama assumed that the probiotic organism they just had isolated from green malt might immensely benefit the injured soldiers who fought the war.

That was the time the Probiotic research began, catalyzed by the indications that probiotics may be able to prevent or improve diarrhoea associated with antibiotic use. How accurate they were! The organism has been healing millions of lives since then, as Sankyo Company developed and commercialized the organism as a formulation for human use. The organism isolated was a spore-bearing lactic acid-forming bacteria. Due to their formation of spores, these bacilli have high heat and acid resistance. It was in the year 1973 that Sankyo, Japan transferred this strain, and the fermentation technology to their Indian counterpart Sanzyme limited ( the then Uni-Sankyo ltd.) the first ever Indo-Japanese joint venture.

Prescription Pattern of SPORLAC other than Diarrhoea*


Cholesterol 10%

Immunity 52%

Jaundice 32% Vaginitis 39%

Aphthous Ulcers - 49%

Immunity Jaundice

Aphthous ulcers

Vaginitis

Cholesterol

*(The survey still being carried out across various zones of India)

INTRODUCTION

jaundice, immunity induction, bacterial vaginitis, etc. A survey has been conducted to understand prescription pattern and inclination of doctors for all the applications of SPORLAC, involving doctors across various parts of India. The survey revealed that around 89% of the doctors prescribe SPORLAC in unconventional applications other than diarrhoea and gut problems. Amongst these doctors 52% prescribe for immunity, 49% for Aphthous ulcers, 39% prescribe in cases of bacterial vaginitis, 32% prescribe in cases of Jaundice and about 10% of doctors in cases of hypercholesterolemia. The observation from survey that 80.5% of these doctors prescribe in more than one unconventional application makes SPORLAC a popular probiotic in indications other than gut problems. Internationally about half a dozen brands of this probiotic have been marketed by companies of distinguished repute. In India, SPORLAC is the most prescribed probiotic and has its reach to lakhs of retailers and doctors across the country making it one of the most popular brands.

PORLAC, the first probiotic in India, has been successfully

used to manage GUT problems, and as it evolved SPORLAC

has been advocated for unconventional indications such as

INDEX

1. Birth of SPORLAC 2. Lactobacillus sporogenes Past, Present & Future 3. Description 4. Characteristics of an Ideal Probiotic 5. Benefits of SPORLAC as a Probiotic 6. SPORLAC Characterization 7. Fermentation Characteristics 8. Morphological & Biochemical Characteristics 9. SPORLAC Acid & Bile resistance 10. SPORLAC Applications a. Gastroenteritis b. Aphthous Ulcers c. Recurrent Bacterial Vaginosis d. Hypercholesterolemia e. Immunity f. Urticaria g. Azotemia h. Necrotizing Enterocolitis 11. SPORLAC Mechanisms of Action 12. SPORLAC* metabolites and EPIGENETIC mechanism 13. SPORLAC Clinical Experiences

a. Use of Sporlac in the Treatment of Non-specific Vaginitis 55 b. Hypercholesterolemia 58

8 10 14 15 18 20

c. Preliminary observations on effect of SPORLAC on serum lipid levels in hypercholesterolemic patients 59 d. SPORLAC (L.sporogenes) in Diarrhoea 14. SPORLAC in Other Clinical Conditions 15. Summary of the Nutritional & Therapeutic Benefits of SPORLAC 16. Emerging areas for SPORLAC Research 17. Bacillus coagulans : a viable adjunct therapy for relieving symptoms of rheumatoid arthritis according to a randomized, controlled trial 18. Clinical Studies with LACBON*

63

64

21 22 28 30 32 34 36 38 40 42 44 46

67

68

69 70

19. PROBITECH The Science & Technology of Manufacturing Probiotics 73 20. Citations 21. International Brands 76 80

50 52

Birth of SPORLAC

1949
The health of the Japanese people, at the end of world war II was at its lowest point. Dr.Nakayama, a Japanese physician, first isolates Lactobacillus sporogenes (Bacillus coagulans) on green malt. He believes that these bacteria could be the answer to the problem of dysbiosis running rampant in the Japanese people.

1964
Japanese tested Lactobacillus sporogenes (Bacillus coagulans) on adults, as well as infants, suffering from diarrhoea and constipation. These clinical trials used antibiotics in conjunction with Lactobacillus sporogenes (Bacillus coagulans).

1972
The Japanese Ministry of Health and Welfare approves and Japanese doctors are able to prescribe Lactobacillus sporogenes (B.coagulans) for diarrhoea and constipation.

1973
Erstwhile Sankyo ( now Daiichi Sankyo Co. ltd.) offered formulation and fermentation technology to Sanzyme ( the then Uni-Sankyo ltd.), that led to the first probiotic of India, by brand name SPORLAC.

Lactobacillus sporogenes
Past, Present & Future

identify the same organism. The species L.sporogenes was originally isolated and described in 1933 by Horowitz-Wlassowa and Nowotel now and subsequently reclassified as Bacillus sporogenes. It has been evidenced that B.sporogenes shares the same characters of B.coagulans, and therefore it has been moved into Bacillus coagulans group. Accordingly to the 8th edition of Bergeys Manual of Determinative Bacteriology, spore-bearing rods producing lactic acid, facultative or aerobic and catalase positive are to be classified within the genus Bacillus. While in the global market it is referred to as a food or nutraceutical ,the current status in India as per the regulatory approvals for SPORLAC treat Lactic Acid Bacillus not as nutraceutical, but as a medicine.

ne country and 3 names Hindustan, Bharat and India.

Likewise Lactobacillus sporogenes, Lactic Acid Bacillus

[LAB] and Bacillus coagulans are three references used to

For all practical purposes we shall refer Bacillus coagulans or Lactobacillus sporogenes or Lactic Acid Bacillus as SPORLAC in this compendium.

Description

makes SPORLAC the ideal supplemental Probiotic, protecting the spores from gastric acid, bile, heat, oxygen and antibiotics. SPORLAC can be stored without refrigeration, and remains viable for extended periods of time. After ingestion, the spore coat dissolves harmlessly leaving the spores to multiply. SPORLAC is composed of L-lactic acid producing spores of lactobacilli. There are three types of lactic acid that occur in the form of three isomers (substances with identical molecular structures that have different shapes): L(+) lactic acid, D(-) lactic acid and DL-lactic acid. These bacteria have antagonistic effects on any bacteria that abnormally increase, resulting in restoration of a normal balance of the bacterial flora in the intestine. In addition, once the intestinal bacterial flora is restored to its normal state, the spore-bearing lactic acid forming bacilli are quickly removed from the body without them being able to form colonies. There have been a number of reports of their favorable effects on patients with acute and chronic inflammation of the intestine and also on those with constipation. The intestinal balance of bacterial composition affects the metabolism in the intestine, thus affecting nutrition, the efficacy of drugs, physiological functions, aging, the occurrence of cancer and immunological functions.

PORLAC is a spore-forming Lactobacillus. The spore is

naturally encapsulated in a protective coat consisting of calcium,

protein and carbohydrate complex. It is this encapsulation that

11

As dietary habits have changed dramatically since World War II and society has become more stressful due to changes in the social environments, probiotics were good health supplements, due to their ability to promote the maintenance of an appropriate intestinal environment. An increasing number of reports on the health promoting effects of SPORLAC in yogurt and health drinks. To enhance health, it is essential for the bacilli to reach the intestinal tract alive and intact. Spores produced by SPORLAC presented as tablets / powder form permit the transfer of the bacilli to the large intestine unaffected by gastric and bile acids (unlike vegetative forms, which are vulnerable during transit), making it the choice probiotic for a variety of ailments either singly or in combination with other probiotics.

13

Characteristics of an Ideal Probiotic


Survival Stress tolerance Acid tolerance Bile tolerance Intestinal Residence Surface Proteins Adherence factors Bio-film formation Functionality Immuno-modulation Bacetriocin production Carbohydrate utilization & metabolism Safety Gene transfer potential Antibiotic resistance Putative virulence factor homologs

Benefits of SPORLAC as a Probiotic

requirements of an efficient probiotic. Preparations of SPORLAC in pharmaceutical dosage form such as tablets, capsules, dried granules or powder have the following characteristics : 1. Contain a large number of viable lactobacilli that retain viability during preparation in pharmaceutical dosage forms and during storage before consumption. The spores are thermostable as against viable L.acidophilus cells which may not withstand lyophilization. 2. Survive in gastric secretions and bile of the upper digestive tract and reach the intestine safely. 3. Settle in the digestive tract and produce enough lactic acid and other antagonistic substances to inhibit the growth of pathogenic bacteria, thus exhibiting antibacterial activity. Being sporulated, they germinate under favourable conditions and produce sufficient viable cells which proliferate and perform vital functions as described earlier. In addition SPORLAC are semiresistant and are slowly excreted out of the body (7 days after discontinuation of administration).

linical studies have revealed that SPORLAC can be successfully implanted in the intestine. As explained in an earlier section, SPORLAC satisfies the essential

15

Diagrammatic Representation of the cytological changes that take Place during the formation of the bacterial endospore

Vegetative cell

Chromatin filament

Spore seplum

Spore Protoplast

Cortex formation

Coat formation

Maturation

upto 1.5 1.5 to 2.5 2.5 to 4.5 4.5 to 6.0 6.0 to 7.0 hours after hours after hours after hours after hours after start of start of start of start of start of sporulation sporulation sporulation sporulation sporulation

SPORLAC is effective in the form of dietary supplements as well as when added to food products. Natto is a traditional fermented product from soya bean, consumed widely in japan as a rich source of protein. Its flavor is improved by the incorporation of lactic acid bacteria such as SPORLAC, L.acidophilus or Pediococcus acidilactiti to the starter culture (Bacillus natto), to yield a product called Yogurunatto having superior flavor and storage characteristics as well as improved nutritional and therapeutic properties. A nutritive medium (homogenized mushroom, Lentinus edodus) is mixed with soya beans and fermented to yield this food product. The requirement of availability of a number of viable lactic acid bacteria is fulfilled by using SPORLAC.

17

SPORLAC Characterization

Multiplication of SPORLAC begins approximately four hours after ingestion and colonies double every thirty minutes, producing beneficial L(+)lactic acid in the small and large intestine through the fermentation of glucose, fructose, sucrose, trehalose and other sugars. SPORLAC is considered a transient inhabitant.

onsequent to the joint FAO/WHO Expert Consultation on evaluation of Health and Nutritional Properties of probiotics in 2001, certain guidelines were framed to set out a systematic approach for the evaluation of probiotics, which include certain in vitro tests to determine physiologic and functional health characteristic of the strain and in vivo trials to substantiate efficacy in humans.

Invitro tests
Acid Bile Stability To exert their beneficial effects probiotics must resist to the acidity of the stomach, lysozyme and bile acid. SPORLAC survives at gastric pH and reach the intestine, where sporulation could occur. Adhesivity Adhesion properties are considered an important issue, and particularly, ability to adhere to intestinal mucosa is one of the essential selection criteria for probiotics, since adhesion to intestinal mucosa represents the first step in colonization process. Stable adhesion to colonic mucosa seems associated to shortening of diarrhoea, immunogenic effects, competitive exclusion and other effects. Bacteriocin Production A plasmid-encoded bacteriocin-like inhibitory substance, named Coagulin, is produced by SPORLAC. Because of its spectrum of activity encompassing other SPORLAC strains, enterococci, Listeria spp, it has been proposed as an alternative to Nisin.

19

Fermentation Characteristics
S. No 1 2 3 4 5 6 7 8 9 10 11 Substrate Glucose Xylose Fructose Galactose Lactose Saccharose Maltose Starch Inulin Glycerol Mannitol Acid Production + + + + + + + + + + +

SPORLAC

Antibiotic Resistance Pattern


S. No 1 2 3 4 5 Antibiotic Cefoxitin Colistin
(Methane Sulphonate)

Resistance pattern + + + + +

Polymyxin-B Novobiocin Metronidazole

Morphological & Biochemical Characteristics


S.No.
1 2 3 4 5 6 7 8 9 10 11 12

Parameter
Vegetative cells Size Motility Endospore Shape

Characterstic
Rods, occurring singly, rarely in short chains, variable in filaments by cultural conditions 0.9 by 3.0 to 5.0 Microns Motile Spore former, oval and Terminal position. Present after 48 hrs incubation at 37o C Ellipsoidal 0.9 to 1.2 by 1.0 to 1.7 microns terminal,occurring single Gram stain - Positive Spore staining - Spores stain red and vegetative cells stains blue. Medium: PNY, Colony shape: Circular Surface: Smooth Edge : Entire without exception Opaque Growth is abundant in ( PNY broth with NaHCO3 0.15%, Pancreatin 0.25 % pH 7.2 incubation 24 hours at 37o C ) pH 4.4 - 4.6 Not produced Not produced

Reaction to stain Surface colonies on solid medium Opacity Growth in simulated intestinal fluid Litmus milk Production of indole Production of hydrogen sulphide Voges proskauer test Acetyl methanol carbinol Catalase Lactic Acid Production

13

Not produced

14 15

Positive Positive

21

SPORLAC
Acid & Bile resistance

forming lactic acid producing bacteria (SFLAB) as probiotics. Due to their spore forming ability, these bacilli have high heat and acid resistance. Most cells of ordinary Lactobacilli die at 70C, while it has been reported that spore-bearing lactic acid forming bacteria do not show a decrease in viable cells even after heating in saline at 85C for 30 min and are also stable in artificial gastric juice (pH 2.0) for a typical time of application (3 hr). SFLAB are a group of Gram-positive bacteria, sharing characteristics common to the genera Bacillus (spore forming, motile) and Lactobacillus (micro-aerophilic, lactic acid production). Many factors make SFLAB good candidates for a probiotic use : (I) They are easily cultured in bulk

number of peer-reviewed publications have cited, that

spores are therapeutically beneficial as compared to

vegetative cells. Studies have been carried out on spore

(II) They produce organic acids (III) Due to spore form, SFLAB are more resistant to heat, which facilitates the pelleting process used in the mass. (IV) SFLAB is resistant to the acidity of the stomach, lysozyme, bile and pancreatic enzymes. Bacterial spores are produced in nature as a means to survive extreme environmental conditions enabling long-term survival in conditions

23

The Bacillus Spore


Cortex Inner Coat Exosporium Nuclear material

Outer Coat Plasma membrane

Cortical membrane of germ cell wall

Incorporated mother cell cytoplasm

Core or protoplast

that could otherwise kill vegetative bacteria. The survival of spores depends on the sporulation during the bio-mass production, when the vegetative forms go into the spore stage. The decision to sporulate is very much dependent upon the decline in nutrients in the immediate vicinity of the live cell. Sensing this, the bacterium enters an irreversible program of development that results in the production of a spore. About eight hours later intrinsic to survival is the structure of the bacterial endospore, that contains, at its core, a condensed and inactive chromosome. Additional layers surround the spore, including a peptidoglycan rich cortex and one or more layers of proteinaceous material referred to as the spore coat, meant to protect the spore from UV radiation, extremes of heat (typically up to 80-85oC in most species), exposure to solvents, hydrogen peroxide and enzymes such as lysozyme. The spore itself, is dehydrated and if exposed to appropriate nutrients will germinate, a process taking just a few minutes, allowing water to enter the spore, breakage and removal of the spore coats, and outgrowth and resumption of vegetative cell growth. The nutrients of the GI tract in the distal end beginning in the ileum and caecum provide ambient environment / conditions and nutrients for the spores to germinate and exert the beneficial effects to the host, a purposeful and healthy symbiotic relationship. Depending on species spores are spherical or ellipsoidal in shape, between 0.8-1.4 mm in length, have a negative surface charge and are moderately hydrophobic.

25

SPORLAC - Life Cycle

Sporulation
MC MC F S F
Starvat ion

Vegetative Cell Growth


Binary Fission

VC
Germination

VC

VC

MC - Mother Cell F - Forespore VC - Vegetative Cell S - Spore

Under conditions of nutrient starvation the growing Vegetative Cell (VC) will undergo a series of morphological changes that create a Forespore (F) within the Mother Cell (MC) of the sporangium. After approximately eight hours the Spore (S) is released by lysis of the MC.

The next question is how do spores germinate, & how is it defined ? Spore germination, is defined as those events that result in the loss of the spore-specific properties, is an essentially a controlled natural or biophysical process during manufacture. It occurs without any need for new macromolecular synthesis, so the apparatus required is already present in the mature dormant spore. Germination in response to specific chemical nutrients requires specific receptor proteins, located at the inner membrane of the spore. The full molecular details of the signal transduction process in spore germination are not yet clear, but reasonable hypotheses can be constructed with the available information, most of which is derived from studies with Bacillus subtilis, the genetic and biochemical paradigm spore-former.

27

SPORLAC
Applications

imbalance (dysbiosis), and probiotics are quite well studied for their proven clinical efficacy with their enteric approach. Following are the applications for which SPORLAC is understood to be useful : 1. Gastroenteritis 2. Aphthous ulcers 3. Recurrent Bacterial Vaginosis 4. Hypercholesterolemia 5. Immunity 6. Urticaria 7. Azotemia 8. Necrotizing Enterocolitis

PORLAC has a wide range of applications, apart from being

conventionally a solution for gut problems. For various novel

applications, there is an etiology that is linked to gut microflora

29

GASTROENTERITIS

Inflammation

Tolerance

SPORLAC Allergen

Balanced microflora

Infection Pathogen Gut barrier dystunction Aberrant microflora

Immune response

Aetiology & Role of SPORLAC

The common Aetiological factors for gastroenteritis include: i) Infections ii) Radiation iii) Antibiotic therapy iv) Tube feeding v) Bacterial agents vi) Viruses These pathogens produce different types of toxins that can cause severe or life-threatening dehydration and diarrhoea.

Gut Microflora in Inflammation Inflammation may direct the composition and function of a balanced normal microflora to become aberrant and immunogenic, leading to perpetuation of the inflammation and gut barrier dysfunction. Probiotics like SPORLAC may 1. Counteract the inflammatory process by enhancing the degradation of enteral antigens, 2. Reduces the secretion of inflammatory mediators, 3. Promote the normalization of indigenous flora and the exclusion of pathogens.

Source: E Isolauri, P V Kirjavainen, S Salminen, "Probiotics: a role in the treatment of intestinal infection and inflammation" Gut 2002;50(Suppl III):iii54iii59.

31

APHTHOUS ULCERS

SPORLAC

Microbiota

Immune deficiency

Stress

APHTHOUS
GIT Disease

ULCERS

Acidic fruits

Vit- B12, Zinc & Folic acid

Sharp tooth surface

SPORLAC

Aetiology & Role of SPORLAC

phthous ulcers or Recurrent Aphthous Stomatitis (RAS) or canker sores, are inflammatory lesions of the mucous lining of the mouth which may involve the cheeks, gums, tongue, lips, and roof or floor of the mouth. Possible aetiological factors include : 1. Trauma & Stress 2. Systemic diseases and Nutritional deficiencies 3. Food allergies 4. Infection 5. Immune disorders 6. Drug induction Antibiotics & Anti-inflammatory Drugs The ability of SPORLAC to increase the activity of the phagocytes must be the key factor in combating RAS, as Porter and Scully have stated that in RAS, phagocytosis by PMNLs (Polymorphonuclear leukocytes) and the chemotaxis to PMNL s are defective. SPORLAC is very useful in Aphthous stomatitis and Glossitis, which may be a result of Vitamin B Complex deficiency due to denudation of intestinal saprophytic flora. According to Dr.S.N.Mathur et.al., SPORLAC is proved most efficacious in the treatment of Aphthous ulcers within 2-3 days. According to Dr.Sharma et.al., SPORLAC therapy is one of the best available methods to treat these recurrent oral ulcerations successfully.

Source: UP State Dental Journal, Vol. II 7-12, 1970.

33

RECURRENT BACTERIAL VAGINOSIS

Replacement of lactobacilli Vaginal or oral

Reducion of Lactobacilli Reduction in H2O2 Production

Overgrowth of BV associated bacteria

Raised pH

SPORLAC
Intermittent treatment Vaginal or oral Maintains pH at 4.5 by vaginal acidifying gel

Aetiology & Role of SPORLAC

included in the study. Complete relief from pruritus and discharge was reported in 91% of the treated subjects. These results were attributed to lactic acid produced in the vaginal milieu which inhibited the growth of vaginal pathogens, by SPORLAC. Post menopausal women, had a slower response to therapy but eventually had complete relief.
Wilson J, Sex Transm Infect 2004;80:811.

PORLAC is administered vaginally or orally in women with

non-specific vaginitis twice daily with for 14 days. Women with

severe infections of Trichomonas and Candidias were not

35

HYPERCHOLESTEROLEMIA

Dietary Fat Bile Salts Emulsification SPORLAC Deconjugation of bile salts and interruption of entero-hepatic circulation of bile acids

Absorption SPORLAC Inhibit Cholesterol hydroxymethyl Synthesis glutarate Co A reductase, the rate-limiting enzyme in cholesterol synthesis

Aetiology & Role of SPORLAC

dietary choices. Genetic contributions are usually due to the additive effects of multiple genes, though occasionally may be due to a single gene defect such as in the case of familial hypercholesterolaemia. A number of secondary causes exist including: Diabetes mellitus type 2, Obesity, Alcohol, Monoclonal gammopathy, Dialysis, Nephrotic syndrome, Obstructive jaundice, Hypothyroidism, Cushing's syndrome, Anorexia nervosa, Medications (thiazide diuretics, ciclosporin, glucocorticoids, betablockers, retinoic acid). Probiotics like SPORLAC acts by 1. Deconjugation of bile salts and interruption of entero-hepatic circulation of bile acids 2. Inhibit HMG CoA reductase thereby inhibiting Cholesterol synthesis 3. Improve elimination

ypercholesterolemia (elevated serum cholesterol levels) is

typically due to a combination of environmental and

genetic factors. Environmental factors include obesity and

Source: Gilland .S.E. and Speck .M.L (1977). Deconjugation of Bile acids by Intestinal lactobacilli, Appl.Environ. Microbiol. 33: 15.

37

IMMUNITY

Lumen
(1) Enhancement of the epithelial barrier Mucins and defensins

(3) Inhibition of pathogen adhesion

SPORLAC

SPORLAC

Pathogens

(5) Production of anti-microorganism substances Mucus

(4) Competitive exclusion of pathogenic microorganisms

(2) Increased adhesion to intestinal mucosa IECs

DCs Lamina propria

SPORLAC
IL-10 TGF (6) Modulation of the immune system Macrophage

Immature DC

Th1 Treg

Th2 Th17

Role of SPORLAC

he immune system can be divided between the innate and adaptive systems. The adaptive immune response depends on B and T lymphocytes, which are specific for particular antigens. In contrast, the innate immune system responds to common structures called Pathogen Associated Molecular Patterns (PAMPs) shared by the vast majority of pathogens. The primary response to pathogens is triggered by Pattern Recognition Receptors, which bind PAMPs. In addition, extracellular C type Lectin receptors and intracellular Nucleotide binding Oligomerization Domain containing protein like receptors are known to transmit signals upon interaction with bacteria . Major mechanisms of action of probiotics like SPORLAC include 1. Enhancement of the epithelial barrier 2. Increased adhesion to intestinal mucosa 3. Concomitant inhibition of pathogen adhesion 4. Competitive exclusion of pathogenic microorganisms 5. Production of anti-microorganism substances 6. Modulation of the immune system Probiotic which have the ability to interact with epithelial and dendritic cells (DC) and with monocytes/macrophages and lymphocytes can exert immunomodulatary effect.

Source : Miriam Bermudez-Brito, Julio Plaza-Daz, Sergio Muoz-Quezada, Carolina Gmez-Llorente, Angel Gil Probiotic Mechanisms of Action , Ann Nutr Metab 2012;61:160174.

39

URTICARIA
2

Elevated secretion of TNF- ECP Increased rate of transmural passage of enteral antigens in their undegraded forms

Oral allergen

Allergic inflammation in the gut Imbalance of intestinal microflora composition

Defective generation o f mu c o s a l I g A response to centeral antigens

SPORLAC
6

Aetiology & Role of SPORLAC

There are various suggested targets of probiotic therapy to modulate immune responses to dietary antigens during the course of allergic inflammation in the gut. Probiotic bacteria such as SPORLAC are shown to modulate allergic inflammation by : 1) 2) 3) Altering the immunogenicity of allergens via proteolytic activity Reducing the secretion of inflammatory mediators in the gut Reversing the increased intestinal permeability and enhancing the degradation of enteral antigens 4) 5) 6) Diverting the gut antigen uptake toward Peyer's patches Normalizing the composition of intestinal microflora Enhancing mucosal Immunoglobulin A (IgA) response to enteral antigens - TNF (Tumor Necrosis Factor); ECP (Eosinophil Cationic Protein).
Isolauri E et al. Am J Clin Nutr 2001;73:444s-450s

41

AZOTEMIA

SPORLAC
Portal circulation Colon Free p-cresyl-sulfate
CH3

Kidney

o Proteins Peptides
CH3

S albumin

Tyrosine Tryptophan

p-cresyl Indole
OH N H

Putereffective bacteria

Liver

o S

H N

Gut lumen

Free indoxyl-sulfate

Prevent the formation of p-cresyl-sulfate & indoxl-sulfate in the gut

Albumin bound p-cresyl-sulfate & indoxl-sulfate not efficiently removed by hemodialysis

Aetiology & Role of SPORLAC

Hemodialysed patients have more than 100 times higher aerobic bacteria (putrefactive bacteria) than normal subjects. Probiotics reduce the putrefactive bacteria by creating the acidic environment and normalizes the microflora. Probiotics such as SPORLAC improves nutrition and decreases the toxic substances like DMA (Dimethyl amine), NDMA (Nitroso dimethyl amine) and toxic amines. The intake of SPORLAC effectively restores the disturbed microflora to a normal one by creating the acidic environment which then prevent the carcinogenic aerobacteria like E.coli. Haemodialysis (HD) cannot effectively eliminate protein-bound solutes as opposed to small water-soluble solutes, and the accumulation of protein-bound uremic toxins such as p-cresol and indoxyl sulphate is suggested to be related to complications such as Azotemia and mortality in HD patients. Serum concentration of indoxylsulphate & p-cresyl sulphate is increased markedly in uremic patients with because insufficient renal clearance. A pilot study found that SPORLAC supplementation for 16 weeks attenuated BUN levels & enhanced survival of 5/6th nephrectomized rats while slowing the progress of renal injury induced by reduced nephron mass. Supplementation of probiotics prolonged the life of uremic rats, thereby showing significantly less mortality (p < 0.05) and highest BW gain.

43

NECROTIZING ENTEROCOLITIS

Prematurity

Increased PAF production and decreased PAF degration

SPORLAC

Formula feeding Intestinal ischemia Excess PAF

Bacterial colonization

Increased TLR Expression; activation of TLR

TLR signaling Production of Pro-inflammatory Cytokines

Breach of mucosal barrier integrity

Bacaterial translocation

SPORLAC

Aetiology & Role of SPORLAC

Necrotizing Enterocolitis (NEC) is a medical condition primarily seen in premature infants where portions of the bowel undergo necrosis (tissue death). NEC has no definitive known cause. An infectious agent has been suspected, as cluster outbreaks in neonatal intensive care units (NICUs) have been seen, but no common organism has been identified. Prematurity, Intestinal ischemia and Bacterial colonization leads to increased production and decreased degradation of Platelet Activating Factor (PAF). Excess of this PAF increases the activation and expression of TLR's (Toll Like Receptors), increased signaling of TLR causes Bacterial colonization, production of proinflammatory cytokines, Breach of mucosal barrier integrity. All these leads to Necrotizing Enterocolitis in infants. Probiotics offered as nutritional supplements that act in the intestine of the host organism by regulating the local bacterial flora. They act by improving gastrointestinal permeability and increasing the resistance of the mucosa against bacterial penetration. Regarding the protection mechanisms, they: (i) Increase the resistance of the intestinal barrier against the passage of bacteria and their toxins (ii) modify the host response in relation to microbial products (iii) increase the mucosal response to IgA (iv) produce bactericidal substances and (v) competitively exclude potential pathogens.

45

SPORLAC
Mechanisms of Action

generate intense metabolic activity, mainly in the colon, and play an important physiological role in the host. The metabolic activities, that exert therapeutic benefits are Proteolysis, Lipolysis and in addition to these activities SPORLAC induce enzymes -galactosidase, Glycolase and Lacatate dehydrogenase. Lactic Acid Production The ability of conversion of lactose to lactic acid, has been used with great success in treating lactose intolerance. Individuals with lactose intolerance, are unable to metabolize lactose due to a deficiency of essential enzyme Lactase. Lactic acid, by lowering the pH of the intestinal environment to 4.0 or 5.0, inhibits the growth of putrefactive organisms and E.coli which survive in optimal pH of 6.0 to 7.0. Volatile acids produced during fermentation, also possess some antimicrobial activity, under conditions of low oxidation-reduction, potential. Production of Bacetriocins SPORLAC is generally recognized as safe (GRAS) and the preservative effect exerted by SPORLAC is mainly due to the production of organic acids (such as lactic acid) which result in lowered pH. It also produces antimicrobial compounds including hydrogen peroxide, CO2, diacetyl, acetaldehyde, D-isomers of amino acids, reuterin and bacteriocins.

irst, it should be understood that the microflora in the adult

human body consist of an enormous biomass of more than

100 trillion bacteria of more than 400 different species, which

47

The metabolites of SPORLAC, that exert antagonistic action against putrefactive micro-organisms and the action of some of the metabolites are summarized

Metabolic Product

Mechanism of Action Inhibits decarboxylation

Carbon dioxide Reduction of membrane permeability Diacetyl Hydrogen peroxide / Lactoperoxidase Interaction with arginine binding proteins Oxidizes basic proteins

Lactic acid

Undissociated lactic acid penetrates the membrane, lowering the intracellular pH. It also interferes with metabolic processes such as oxidative phosphorylation. Affects membrane, DNA -

Bacteriocins

Synthesis and protein synthesis

Bacteriocins are ribosomally synthesized antimicrobial peptides that are active against other bacteria, either of the same species (narrow spectrum), or across genera (broad spectrum). In recent years, bacteriocin producing probiotics have attracted significant attention because of their GRAS status and potential use as safe additives for food preservation. Some of the bacteriocins, produced by SPORLAC are Acidolin, Acidophilin, Lactain B & F. Vitamin-B Synthesis Mammalian cells cannot synthesize folate (an important B-group vitamin, participates in many metabolic pathways such as DNA and RNA bio-synthesis and amino acid inter- conversions); therefore, an exogenous supply of this vitamin is necessary to prevent nutritional deficiency. Milk is not a rich source of dietary folate. However, many dairy products are processed using microbial fermentations in which folate can be synthesized, significantly increasing folate concentrations in the final product. Numerous researchers have reported that SPORLAC, has the ability to synthesize folate.

SPORLAC confer physiological benefits to the hosts: 1. Improving digestive capacity of the consumed milk proteins 2. Improving absorption and mineral metabolism [Calcium, Phosphorus & Iron] 3. Stimulates secretion of gastric juices 4. Improving peristaltic movements of the digestive tract 5. Serving as an energy source in cellular respiration

49

SPORLAC* metabolites and EPIGENETIC mechanism

Probiotics Prebiotics + Trophic and anti-neoplastic effect Reduced colon cancer risk and IBD inflammation

Fe

rm

en

tat

io

n
SCFA De novo lipogenesis

Antagonism of pathogens and putrefactive bacteria

Reduced pH Ca++Mg++
Increased mineral absorption

Controlled serum lipids and cholesterol HDAC inhibitor

Fewer toxic bacterial metabolites Epigenetic regulation

Reduced cancer risk

Colonocytes

Probiotics when ingested orally ferment indigestible carbohydrate from food and produce short chain fatty acids (SCFA). Anaerobic bacteria also produced SCFA from peptide, polysaccharide, protein and oligosaccharide and are the final product of bacterias activity into the GI tract. SCFA can lower the lipids in blood through blocking synthesis of hepatic cholesterol and/or through redirecting plasma cholesterol toward the liver SCFA are potential modulator of food intake and energy sensing process into the brain, which might indirectly play an important role in reduction of cholesterol and other metabolism deranging lipids into the host body. SCFA involve reduction of pH which is important phenomenon for increased absorption of Ca, Mg minerals and reduced pH results in fewer toxic bacterial metabolites thus reduces cancer risk. Epigenetic Regulation: SCFA effects on HDAC ( Histone Diacetalyse ) inhibitors, a class of compound that interfere with the function of HDAC inhibition will have a effect on psychiatry and neurology as mood stabilization and epileptics. And this concept investigated for possible treatments for cancer and inflammatory diseases like IBD.

51

SPORLAC
Clinical Experiences

Clinical Trials In The Treatment of Aphthous Stomatitis & Glossitis


Mathur, S.N. et al. (1970). UP State Dental Journal 11:7-12

Sharma et al., noted that SPORLAC therapy is one of the best available methods to treat recurrent oral ulcerations. At the dosage level of two tablets thrice daily for five days, (corresponding to 120 million spores of L.sporogenes per day), aphthous stomatitis was cured in two to three days.

Lactic Acid Bacteria Is Beneficial For Oral Aphthous Ulcerations - A Preliminary Report
Hashim, B.Y; Rahman, R.A; Philip, K. Annal Dent Univ Malaya 1999; 1: 43 _46

Recurrent aphthous ulcers of the mouth are difficult to treat because of no known definite aetiology. This paper presents the use of lactic acid bacteria thought to modulate the host immune response to affect improvements in the disease. Twenty-five patients with the disease were treated with six lactic acid bacteria capsules daily for a period of six months, and their responses were evaluated. Seventeen patients (73.9%) became free of the disease six months later, while six (26.1 %) experienced very dramatic improvements. Two patients were lost to follow-up. It is concluded that lactic acid bacteria is beneficial in the treatment of recurrent aphthous ulcers of the mouth.

53

Acidic Alkaline
Change in Vaginal pH with Treatment

45 40
Number of Subjects

35 30 25 20 15 10 5 0 0 3 5 7 14
Duration of Treatment (Days)

Complete relief of pruritis and discharge was reported by 91 percent of subjects. These results were thought to be due to a beneficial change in vaginal acidity via lactic acid production by SPORLAC. Postmenopausal subjects had a slower response to therapy but eventually had complete relief as well.

Use of SPORLAC in the Treatment of Non-specific Vaginitis


Shirodkar NV, Sankholkar PC, Ghosh S, Nulkar SM. Indian Pract 1980;33:207-210.

Non-specific vaginitis is caused by a variety of pathogens including Staphylococci, Streptococci, Pneumococci and E.coli. It may also be induced by a variety of causes including chemicals, drugs, surgical procedures, trauma and foreign bodies. SPORLAC administration to increase the vaginal acidity by the action of the Lactobacillus on glycogen in the vaginal epithelial tissues was adopted in a clinical trial on 44 patients. The patients were divided into two groups: Group 1: Twelve patients suffering from leucorrhea (white discharge) following cervical surgery. Group 2 : Thirty two patients with nonspecific vaginitis without previous therapy. Of these, 26 were in the reproductive age and 6 were menopausal. The change in vaginal pH following treatment with SPORLAC tablets over a period of two weeks is shown in the figure:

Most of the cases showing persistently alkaline pH were postmenopausal, where acid could not be produced in sufficient amounts due to low substrate glycogen levels. Glycogen levels depend upon circulating estrogen.

55

The Response to Treatment in Both Groups

Number of Cases
Type of Response Quick Response & Complete Relief Group 1 8 (67%) Group 2 26 (81.25%) Total 34 (77.25%)

Delayed Response But Complete Relief

2 (16.5%)

4 (12.5%)

6 (13.60%)

Improvement But Not Complete Relief

2 (15.5%)

2 (6.25%)

2 (9.15%)

No Relief

0 12 (100%)

0 32 (100%)

0 44 (100%)

Total

In comparision with a clinical trial using M.T.P. vaginal pessaries containing broxyquinoline and brobenzoxeldine where only 26.67% of cases studied were cured, it can be seen that SPORLAC therapy is the better alternative in the treatment of non-specific vaginitis. SPORLAC therapy provided complete relief to 91% of the patients and partial relief to approximately 9%. Vaginal administration of a commercial formulation of SPORLAC tablets called MYCONIP was given to 44 women with non-specific vaginitis twice daily for 14 days. Subjects with Trichomonas or Candida vaginitis were excluded from the study. Complete relief of pruritis and discharge was reported by 91 percent of subjects. These results were thought to be due to a beneficial change in vaginal acidity via lactic acid production by SPORLAC. Postmenopausal subjects had a slower response to therapy but eventually had complete relief as well.

57

Hypercholesterolemia

Hypocholesterolemic Effects of SPORLAC : Clinical Studies

350 300

Serum levels mg/dl

Before After

250 200 150 100 50 0 Total Serum Cholesterol (p,0001) LDL Cholesterol (p<0.001) HDL Cholesterol (p<0.05)

Preliminary observations on effect of SPORLAC on serum lipid levels in hypercholesterolemic patients.


Mohan JC, Arora R, Khalilullah M.Indian J Med Res. 1990 Dec; 92:431-2.

Short term hypolipedemic effects of oral SPORLAC therapy (369 million spores per day in tablet form) were studied in 17 patients (15 men and 2 women in the 32-61 year age group) with type II hyperlipidemia in an open label fixed dose trial. Total serum cholesterol, LDL-Cholesterol and total cholesterol to HDLCholesterol and LDL-Cholesterol to HDL-Cholesterol ratios (p<0.001) was reduced significantly over a period of three months. HDL-Cholesterol was marginally increased (437mg/dL vs 46.88.9 mg/dL, p<0.05), there was however no change in serum triglyceride levels. Atherogenic lipid ratios observed were as follows Total/HDL-Cholesterol 24.0% decrease, LDL/HDL-Cholesterol 33.4% decrease. No adverse effects of therapy were noted, except constipation in one patient.

59

The Basic Mechanism of Action of SPORLAC as hypolipidemic agent

Cholesterol Precursor of Bile Salts

De-Novo Synthesis Conjugated Bile Salts Taurine - Conjugated Bile Lactobacillus Glycine - Conjugated Bile Salts Lactobacillus

Bile Salt Hydrolase Hydrolysis

Amino Acid Group Free Cholic Acid Taurine Reabsorbed into GIT Free Cholic Acid Glycine

Excreted into the Faeces

Cholesterol-Lowering Effects of Probiotics and Prebiotics: A Review of in Vivo and in Vitro Findings Lay-Gaik Ooi and Min-Tze Liong Int. J. Mol. Sci. 2010, 11, 2499-2522

The basic mechanism is to prevent the absorption of fat and cholesterol in the form of micelle for which bile salts are essential in the small intestine. Bile salts and cholesterol have enetro-hepatic circulation and any method of interfering in this process will hinder their absorption and consequently the serum levels. Bile salts are deconjugated in the small intestine by the bacterial flora to a small extent and these deconjugated salts can not be incorporated in the micelle. Implantation of large quantities of Lactobacilli to facilitate this deconjugation process is an attractive alternative method to treat hyperlipidemias but has not been tried so far. Availability of spores of SPORLAC which germinate the vegetative form in the gut after oral intake has solved this problem.

61

The Symptoms Experienced by Subjects in Addition to Diarrhoea

Jaundice Septicemia Cord Infection Vomiting

3 3 3 3

Results of Diarrhoeal Symptoms

Condition Diarrhoea Constipation Jaundice

Cases Treated 60 3 3

Cases Cured 49 3 3

Success Rate 81.7% 100% 100%

SPORLAC (L.sporogenes) in Diarrhoea


Dhongade, R.K. Anjaneyulu .R Abstract from Maharastra Medical Journal

A total of 60 cases of diarrhoea with watery stool frequency greater than 6 were examined for efficacy of SPORLAC treatment. Based on the suggested dosage level of SPORLAC at 5 million spores per kilogram body weight, each neonate was given a spore level of about 15 million spores per day. Most of the subjects (about 80%) had a history of breast-feeding. About 19% were both breast and bottle-fed and 1% were bottle-fed. The average duration for recovery was 1.8 days.

As compared to the normal practice of administration of antibiotic and anti-diarrhoeal mixtures, the complicating side effects were not seen in the series of SPORLAC trials. The average recovery time of 1.8 days helped to reduce dehydration in the subjects to a great extent. In a similar study in Japan a comparable success rate of 78.4% with SPORLAC treatment for infantile diarrhoea was obtained. An earlier study in India by Mathur et al. Found the average time for improvement in diarrhoeal conditions to be two to three days, with treatment.

63

SPORLAC
in Other Clinical Conditions

SPORLAC & Lactose Intolerance Individuals with a deficiency of the enzyme -galactosidase [lactase] suffer from abdominal distress, such as diarrhoea, flatulence, bloating etc, when they consume milk and milk-products. However they are tolerant to Yogurt enriched with SPORLAC. It was found to have considerable lactase activity when tested in vitro. Lactobacilli provide considerable amounts of the enzyme Lactase, which hydrolyzes the lactose. The hydrolyzed lactose is converted to physiological amounts of lactic acid. SPORLAC & Antibiotic Associated Diarrhoea Treatment with antibiotics, has been associated with diarrhoea, due to the imbalance of the GI microflora. The main causative organism is C.difficile, which causes diarrhoea even after discontinuation of the antibiotic. Treatment with SPORLAC, has replenished the intestinal microbiota, and causing a reduction in the frequency and duration of diarrhoeal episodes on administration of either powder or tablet form. In infants and neonates, there was significant reduction in the number of episodes of diarrhoea due to viral infections and also improved the feeding pattern, when administered with ORS.

65

SPORLAC & Intestinal Infections Dysbiosis, causes an imbalance in the gastrointestinal milieu may occur due to antibiotic use, immune suppression, allergy insult and stress. In order to achieve, optimal balance between beneficial bacteria and pathogenic bacteria, administration of probiotic microorganisms helped alleviate the abdominal distress in a variety of clinical conditions. The Summary of studies are cited in table.

SPORLAC & Other Associated Disorders

IBS [Irritable Bowel Syndrome] A multifactorial disorder, with varied aetiology of episodic abdominal pain, distension, flatulence, alternating episodes of diarrhoea and constipation, symptoms have shown improvement on administration of SPORLAC.

Oral Conditions Apthous ulcers and Stomatitis SPORLAC administered in doses of upto 120 CFU/day resolved the symptoms of apthous ulcers and stomatitis in as little as 2-3 days

Dental caries SPORLAC was administered to children in whom salivary Streptococcus mutans was detected. Treated individuals were tested for Streptococcus and there was a significant reduction, suggesting that SPORLAC formulations in paediatric population may be effective in preventing dental caries.

Summary of the Nutritional & Therapeutic Benefits of SPORLAC


Lactobacilli
Therapeutic Benefits Nutritional Benefits

Restoration of the Ecological Balance of the GI Flora Detoxification Elimination of end products with carcinogenic potential Inhibition of Growth of pathogens Alleviation of Lactose Intolerance Enhancement of Immunity 1. Vitamin B & Folate Production 2. Improved Digestive Capability of ingested food-stuffs 3. Improved bio-availability of minerals and trace elements

67

Emerging areas for SPORLAC Research


The importance of the microbiota organisms & the manifetstions
Gut-brain hypothesis 1. Autism C. Bolteae/ clostridia spores mechanism unknown 2. Mood: depression, anxiety Asthma/atopy hygeine hypothetic Exagerated innate immune response up regulation of regulatory T cells after capture of Ags by DCs Bifidobacteria, gram +ve organisms clostridia Hypertension/ Ischemic heart disease Billary disease Altered enterohepatic circulation of bile

Diet high in red meat and animal fat Low SCFA/butyrate High fecal fats Low vitamin absorption 7a dehydroxylating bacteria: cholic acid deoxycholic acid (cocarcinogen) Colon cancer Low in H2S metabolizing acteria

Altered xenobiotic/ drug metabolism e.g. paracetamol metabolism: predose urinary p-cresol sulfate leads to postdose urinary acetaminophen sulfate: acetaminophen glucuronide. Bacterially mediatedp-cresol reduces the effective systemic capacity to sulfonate acetamtnophen.

Obesity/metabolic syndrome Peripheral vascular disease result of metabolic syndromealtered lipid deposition/ metabolism Bacteroidetes and Actinobacteria in obese Altered energy/lipid metabolism Higher relative abundance of glycoside hydrolases, carbohydrate-binding modules, glycosyltransferases, polysaccharide lyases, and carbohydrate esterases in the Bacteroidetes TLR mediated Inflammatory bowel disease Hygiene hypothesis Altered immune response:TLR signaling Less microbial diversity Activation of specific species e.g., Escherichia

Diseases influenced by gut microbial metabolism. The variety of systemic diseases that are directly influenced by gut microbial metabolism and its influence on other mammalian pathways, such as the innate immune system, are shown. Specifically highlighted are the metabolic pathways involved in drug metabolism and obesity that are directly influenced by the gut microbial content. Ags, Antigens; Clostridium bolteae; Dendritic cells; SCFA, (short-chain fatty acid) TLR, Toll-Like Receptor. Kinross et al. Genome Medicine 2011 3:14. [Gastroenterology Research and Practice Volume 2012, Article ID 872716, 16 pages]

Bacillus coagulans : a viable adjunct therapy for relieving symptoms of rheumatoid arthritis according to a randomized, controlled trial
Mandel DR,Eichas K and Holmes J BMC Compl. & Alternat. Med. 2010, 10:1

The anti-inflammatory and Immuno-modulating properties of SPORLAC and other lactic acid bacteria, have shown an impact on the symptoms of arthritis. In randomized studies, men and women with an average age of 62 years, received SPORLAC of 2 Billion CFU for 60 days, in addition to their regular arthritis medications. Evaluations of pain and disability at 30 and 60 days of the treatment, showed statistically significant reduction in pain scale scores and disability as compared to placebo. A reduction in c-reactive protein was seen, in the treatment group but not the placebo group. This development coupled with well controlled studies could be just the beginning of a new era, wherein probiotics could call the shots in not just acute but chronic disorders and also the future may see the development of probiotics as targeted carriers of vaccines, hormones and other bio-therapeutic agents.

69

Clinical Studies with LACBON*


Diagnosis Number of subjects Treatment: Lactobacillus spores (millions)
100-600/day in divided doses for 2-12 days

Effective Rates

Remarks
Recovery from diarrhoea to regular normal stools; general symptoms including anorexia improved. Recovery from diarrhoea to regular, normal stools from third to fourth day. Recovery to normal stools & disappearance of abdominal distension. Vomiting & nausea disappeared; appetite improved; stools became normal & regular; diarrhoea & stomach ache cured. General condition & nature of stools improved. Frequency of stools decreased to half or less than that before medication. Obvious eruptions of strophulus and eczema decreased from the third day (topical therapy employed concomitantly). Response seen in anorexia of nervous type and malnutrition in infants.

Acute and chronic intestinal catarrh

38

86.80%

Diarrhoea

15

75-600/day in divided doses for 3-12 days. 300-750/day in divided doses for 2-10 days.

100.00%

Constipation

10

70.00%

Abdominal intestinla fermentation

300-600/day in divided doses for 3-14 days.

100.00%

Dyspepsia Infantum

26

100-200/day in divided doses for 1-7 days.

84.60%

Allergic skin diseases

200-450/day in divided doses for 4-12 days

80.00%

Miscellaneous

10

20-50/day in divided doses for 4-20 days.

80.00%

* is the L.sporogenes / B.coagulans brand of erstwhile Sankyo Japan

Author

Year

Condition
Antibiotic associated diarrhoea Irritable bowel syndrome diarrhoea predominant (IBS-D)

Study Daily Population Dose

Results

La Rosea et al

2003

98 children

71%of children in treatment group had resolution of diarrNot hoea versus 38% in placebo specified group; duration significantly shortened in treatment group. Decreased number of bowel 2 billion movements, no statistically significant change in other cfu IBS symptoms Statistically significant improvements in abdominal pain and bloating in treatment group compared to placebo. Statistically significant decrease in frequency and duration of diarrhoea in treatment group compared to placebo. No effect on rate of death or NEC on VLBW infants; significantly improved feeding tolerance in treatment group

Dolin et al

2009

52 adults

Hun L

2009

IBS-D

44 adults

800 million cfu

Chandra RK

2002

Acute retrovirus 112 newborns diarrhoea

100 million cfu

Sari Fn et al.

2011

Narcotizing enterocolitis (NEC)

221 very-lowbirth weight (VLBW) neonates (<1500 g, <33 weeks)

350 million cfu

Dutta p et al

2011

Diarrhoea with 148 infants dehydration 16-24 months (diverse etiology)

No therapeutic effect on management of acute Not dehydrating diarrhoea of specified diverse etiology, including rotavirus associated diarrhoea in children. Significant reduction in intestinal gas and pain scores; 2 billion strong trend toward improved cfu abdominal distention scores; improved quality of life scores.

Kalman DS et al

2009

Intestinal pain, gas bloating

61 adults

71

Powder production

Bulk dosage forms

Final dosage forms

Boxing & Labeling

Distribution

Store Shelfing

Consumption End of shelf life

potency potency point point #2 #1 Illustrative example of potency decline 6.5B 5.5B

potency point #3

potency point #4

5B

1B

Sanzyme Guarantees CFU count All the way to potency point #4 when delivering to final packaging material

The Shelf-Life Equation

Shelf - Life

Water Activity

Temperature

Oxygen

Shelf - Life Rule-of Thumb Recommendation

24 months

< 0.15a

< 25 C

< 5%

Important Terms:

CFU - CFU = Colony Forming Units ,One CFU = one live bacteria cell [Live defined as being able to multiple and thereby form a colony ] Potency is minimum number of CFU in a Sachet or in one dosage form of either Capsules or Tablets

PROBITECH

TM

The Science & Technology of Manufacturing Probiotics In the early 1973s after the transfer, of the parent culture and the technology to manufacture, SPORLAC was handed over, then came the role of R&D team of Sanzyme. Over the years, Sanzyme has developed a trademarked technical process known as PROBITECH for the manufacture of Probiotics for human application and also those useful in veterinary, aquaculture and bio-remediation. The process of PROBITECH has evolved over 4 decades, as the challenges faced for developing, designing and manufacturing. The Basic Steps in PROBITECH 1. Selection of Strains : Though the task looks seemingly easy, the compatibility of the species if spore forms are combined with vegetative forms or vegetative forms combined with other vegetative forms 2. Purification & Drying : Once the strain/strains are chosen, the fermentation process is decided upon and once the desired strains are isolated, they are purified of impurities and are tested for bacterial contamination and other tests for ensuring that the desired strain is obtained. The process of drying with ambient temperature is esssential, as spore forms withstand the process of drying, but vegetative forms may not be able to withstand the temperatures other than the optimal. 3. Manufacturing : Once the strain/strains are obtained, compatible excipients are chosen to ensure that the spores or vegetative forms do not germinate after manufacture either in the capsule, tablet or sachet

73

PROBITECH

Other

Indication

Regulatory Claim
3

Shelf-Life

Product

Strain/Strains

Potency

Dosage Form

GI Transit Survival

Summary of Probitech Technology


Ingredients Compatibility of Probiotics Excipients & Packaging are crucial Oxygen In combi-Probiotic blends the fine art of maintaining aeration without altering the potency of the blend is crucial Moisture Levels & Water Activity [aw] Crucial Storage of manufactured products can be increased

Physiological State of Bacteria When Prepared in the Product

Temperature [Critical Factor] Manufacture & Storage

pH In-vitro & In-vivo Survival Gastric Transit Time[1-2hrs.] Quiescent Cell-Enhanced Survival

form. During manufacture, correct compression of the tablet without excess pressure and temperature must be maintained for obtaining finished formulations. The blend in the case of combi-probiotic when filled in the capsule must be maitained at the correct temperature while filling and packing into either strips or blister packs. 4. Shipping of Finished Goods : The probiotic either single or the blends are formulated in ideal conditions and ambient temperatures, however as the climate in different geographical regions of India or the globe are not ambient, hence a suitable packaging must be designed to ensure that the probiotic formulations are able to withstand both hot and cold climates. However certain formulations may need to be stored in refrigerated conditions 5. Ensuring Shelf-Life & Potency : In order to ensure that the desired therapeutic benefits are conferred, the desired potency of the probiotic must be maintained to the end of shelf-life and the viability of either the single probiotic or the blend must be maintained as there would be some loses during the manufacturing stages and adequate overages must be added to ensure that label claims are met as per the norms of the drug authorities.
In addition to SPORLAC Powder and SPORLAC DS Tablets, with PROBITECH, innovative blends have been developed in the following categories:

Life Style Cholesterol Busters & Stress Busters Renal Care Prevention of Renal Calculi, Prevention & Treatment of Azotemia Womens Health- Oestrogen recycling, Bacterial Vaginosis and UTIs GI Care Oro-care, Liver Disorders, IBS/IBD,Diarrhoea ENT and Immunity Rheumatoid arthritis Under study

75

Citations
1.Screening of probiotics for acid and bile tolerance was evaluated and B.coagulans strains were tolerant to bile concentrations over 0.3% (w/v) Int (2000):193197 2.One single Bacillus strain is often able to produce several types of molecules stable over a wide range of pH and temperature .these substances are usually protein- and peptidebased compounds such as enzymes, bacteriocins and lipopeptides which are inhibitory compounds, mainly active against food-borne and gut pathogens- Science against microbial pathogens: communicating current research and technological advances FORMATEX 2011 3...The Bacillus coagulans-based product was effective in improving the quality of life and reducing gastrointestinal symptoms in adults with post prandial intestinal gas related symptoms and no GI diagnoses - BMC Gastroenterology 2009, 9:85 4......Immunomodulatory and inflammaotry effects . Lessens the symptoms of arthritis. .. Safe and effective for patients suffering from Rhuematoid arthritis - BMC Complementary and Alternative Medicine 2010, 10:1 5.The unique characteristics of Lactosporin a novel antimicrobial protein produced by Bacillus coagulans, including its antimicrobial activity against pathogenic microorganisms, indicate that it may have potential for application in foods ... - J of Appl Microbi106 (2009) 13701377 6.... results indicate that the administration of B. coagulans . improves the intestinal e nv i r o n m e n t , d e f e c a t i o n f r e q u e n c y, f e c a l c h a r a c t e r i s t i c s a n d d e r m a l characteristics.[reduction in comedones] - Microb. Ecol. in Health and Dis. 2002; 14: 413 7......results of this toxicological safety assessment indicate that B coagulans does not demonstrate mutagenic, clastogenic, or genotoxic effects ... and since the suggested human dose is in the range of 100 x 106 to 3 x 109 CFUs, this gives a safety factor ranging from 3173 to 95,200 times. - Food and Chemi.Toxicol. 47 (2009) 12311238 J of Food Micro; 61

8.....the probiotic strain Lactobacillus sporogenes shows a significant stimulation of the cell mediated immunity and humoral immunity - Int. J Therap. Appl. Vol.3, 2012, 32 38. 9......Probiotic bacteria have a potential role in regulation of intestinal inflammation and also play a role in intestinal immune homeostasi . Commensal bacterial strains, including probiotic bacteria, selected for their properties to activate an epithelial inhibitory response may be a rational nutritional intervention in IBD patients - Eur. J Clin. Nutr. (2002) 56(3):S60S64 10.Perturbation of the intestinal microbiota may lead to chronic diseases such as autoimmune diseases, colon cancers, gastric ulcers, CV disorders, functional bowel diseases and obesity but the use of probiotics has led to promising results by restoring gut flora and alleviating disease - Gastro. Res. and Pract.Volume 2012 11.A one-year chronic oral toxicity study combined with a one-generation reproduction study was conducted to further investigate safety of long-term consumption .is safe for chronic human consumption at an accumulative daily intake of 9.38 x 1010 CFUs per day [human] - Food and Chem. Toxicol. 49 (2011) 11741182 12.Probiotics will provide a safer way to treat the once before incurable, deleterious ailments like cancer It will not be an exaggeration to say that probiotics in the future will be the reigning therapeutic agents - J Res. in Biol.(2012) 2: 102-113 13..... to aid in the digestion of lactose and fructose could be used to prevent occurrence of intestinal symptoms in individuals sensitive to these carbohydrates and in patients with pancreatic exocrine deficiency disorders such as chronic pancreatitis and cystic fibrosis, in which only small amounts of the normal amount of pancreatic enzymes are excreted B coagulans could contribute to the digestion of the protein and carbohydrates in their meal - Beneficial Microbes, 2010; 1(1): 31-36 14.The intestinal microflora is a positive health asset that crucially influences the normal structural and functional development of the mucosal immune system ... The flora has a collective metabolic activity equal o a virtual organ within an organ, and the mechanisms underlying the conditioning influence of the bacterial on mucosal homeostasis and immune responses are beginning to be unraveled for treating ... several infectious, inflammatory and neoplastic disease processes- EMBO Reports (2006) 7, 688693 rld.

77

15. A serious problem of effective pro-biotic usage is the survival of the microorganisms in the stomach and intestines after administration. If spores are used as probiotics instead of vegetative cells, the survival and subsequent inoculation levels might be much higher ..the higher resistance to external factors such as toxic compounds, extremes of temperature, mechanical force etc.... - TRENDS in Biotech. Vol.21 No.8 Aug. 2003 16. Babar V, Thomas R and Bhaskar M, " Immunomodulatory Activity of Lactobacillus Sporogenes ( Sporlac )." Int.l Jl of Therap. Appl. Vol.3, 2012, 32 - 38. 17. Yueh-Ting Tsai & Po-Ching Cheng & Tzu-Ming Pan " The immunomodulatory effects of lactic acid bacteria for improving immune functions and benefits." Appl Microbiol Biotechnol (2012) 96:853862. 18. Sharma, J.K , Kapoor, K.K , Mukhija R.D, " Clinical Trial of Sporlac in the Treatment of Recurrent Apthous Ulceration." U.P State Dental Journal Vol. 11, January 1980, 7-12. 19. Mathur, S.N. et al." Clinical Trials In The Treatment of Apthous Stomatitis & Glossitis." UP State Dental Journal ; (1970).11:7-12. 20. Hashim, B.Y; Rahman, R.A; Philip, K, " Lactic Acid Bacteria Is Beneficial For Oral Aphtous Ulcerations - A Preliminary Report." Annal Dent Univ Malaya; 1999; 1: 43 _46. 21.Wilson J, " Managing recurrent bacterial vaginosis."Sex Transm Infect 2004;80:811. 22.Shirodkar NV, Sankholkar PC, Ghosh S, Nulkar SM " Use of Sporlac In The Treatment of Non-specific Vaginitis." Indian Pract. 1980;33:207-210. 23.Dr. R.C. Arora , " To Evaluate The Role of Sporlac (Lactobacillus Sporogenes) In Jaundice Patients." MLB Medical collage Jhansi. 24.J.C.Mohan , M.N.A.M.S, Ramesh Arora , " Short term effects of oral Lactobacillus sporogenes in patients with Dyslipidemias: an open fixed dose trial." Indian Heart J. 1990 Sep-Oct;42(5):361-4. 25.Lay-Gaik Ooi and Min-Tze Liong " Cholesterol-Lowering Effects of Probiotics and Prebiotics:A Review of in Vivo and in Vitro Findings." Int. J. Mol. Sci. 2010, 11, 24992522.

26.Mohan JC, Arora R, Khalilullah M "Preliminary observations on effect of Lactobacillus sporogenes on serum lipid levels in hypercholesterolemic patients." Indian J Med Res.990 Dec;92:431-2. 27.Dhongade, R.K., Anjaneyulu, R. (1977). Abstract from Maharashtra Medical Journal Vol.XXIII NO.1, Feb: 473-474. 28.Mandel DR,Eichas K and Holmes J " Bacillus coagulans: a viable adjunct therapy for relieving symptoms of rheumatoid arthritis according to a randomized, ontrolled trial." BMC Compl. and Alternat. Med. 2010, 10:1. 29.Katsutoshi Ara, Shinichi Meguro, Tadasi Hase et al. " Effect of Spore-bearing Lactic Acidforming bacteria (Bacillus coagulans SANK 70258) administration on the Intestinal Environment, Defecation Frequency, Fecal Characteristics and Dermal Characteristics in Humans and Rats." Microb. Ecol. in Health and Dis. 2002; 14: 413. 30.Gandhi, A.B. (1988) Lactobacillus sporogenes, an advancement in Lactobacillus therapy. The Eastern Pharmacist, 41-43. 31.Naurse, K and Naruse, W. (1978). Method for producing natto containing lactic acid bacteria. U.S. Patent 4,110,477. 32.Brahman. J. et.al (1969) "L. Acidophilus in the prevention of recurrent headaches." Harefuah 76: 201-202. 33.Bloksma. N. et.al (1981) "Effects of lactobacilli of parameters of non-specific resistence in mice" Medical microbiology and Immunology.170: 45-53. 34.Kishidia .T "Interferon and Immune function" New editions Healt world.

79

SPORLAC Family

Powder Diarrhoea

Tablet AAD Aphthous Ulcers

Powder / Caps Gastroenteritis Bacterial Vaginosis

Powder Immunity

Tablet In Diabetes Zinc & Chromium Deficiency

Powder Acute Infectious / Non Infectious Diarrhoea

Capsule Hyperoxaluria

Capsule Azotemia

Capsule Hypercholesterolemia

The Leader...
Each tablet of SPORLAC Tablet contains 60 million spores needing 6 tablets of dose in a day (2 tablets x 3 times). The launch of SPORLAC DS tablet containing 120 million spores, facilitates convenience of one tablet 3 times in a day.

Sanzyme Ltd.
Healthcare Business

Plot No.13, Sagar Society, Road No.2, Banjara Hills, Hyderabad - 500 034, A.P., INDIA. Tel:+91-40-23553892, 91-40-23554270 / 71, www.sanzyme.com.

You might also like