The word "probiotic" (from Greek – create life) was first introduced in 1954 when harmful effects of antibiotics were compared with the beneficial effects of good bacteria probiotica.
Modern understanding of the importance of probiotics is from the beginning of XXth century due to the Russian biologist and Nobel prize winner, the principal of the Pasteur Institute in Paris prof. Il. Mechnikoff. In his theory "Longevity and rejuvenation of man" prof. Metchnikoff stated that putrefactive processes develop in the human large intestine resulting in toxin production, responsible for the intestinal auto-intoxication that damages all internal organs. Mechnikoff found out that this harmful influence can be eliminated by regular administration of lactobacilli. He arrived at this conclusion after he had found that the long and healthy life of Bulgarian peasants is a result from every day consumption of fermented milk and other lactic acid products made in Bulgaria. At that time the percentage of the centenarians in Bulgaria was the highest (4%), cancer rate and total sick rate were low. Prof. Mechnikoff wrote: "Our aging is a disease that must be treated like any other disease" and recommended every day consumption of lactic acid products. In 1905 the Bulgarian physician St. Grigorov discovered Lactobacillus bulgaricus in these dairy products.
In the recent years it was found out that thousands of microbial strains coexist with people. They are everywhere – on skin, hair, nails, in mouth, intestines, genito-urinary tract. Microorganisms are divided into three groups:
- harmful (pathogenic) – about 150-180 strains;
- beneficial (probiotics) – about 15-20 strains;
- latent (conditionally pathogenic) – over 1000-1200 strains.
Probiotics are live microorganisms which if administered alone or by food, containing probiotics, they heal the human organism.
According to the World Health Organization the total number of the probiotic bacteria with proven beneficial effects is 15. They are given in table 1.
Generally recognized probiotic bacteria used in industry (Table No. 1)
They are divided in three basic groups: lactobacilli, lactic acid cocci and bifidobacteria. According to a Report from 2005 before the Congress of USA, Lactobacillus Bulgaricus is the most important bacterium for people. It is the only probiotic named after a certain geographic area and nation.
Cultivation of Lactobacillus bulgaricus by the Thracians more than 7000 years ago and inventing lactic acid food is one of the three revolutions together with cultivation of crops and domestication of animals that have changed the development of mankind. It was found that some genetic changes occured first in Thracians and then in the other Europeans. The new lactic acid food and the billions of beneficial (probiotic) bacteria created a gene charged with the production of the enzyme lactase which breaks down milk sugar in human body. This new technology granted the rare privilege to people to eat throughout their lives the dairy products obtained from the milk of the domestic animals. People started getting the vital vit. D from the lactic acid products heretofore they have been getting by the sun. This fact made the Thracians and the other Europeans less dependent on the sun shine and they started settling throughout the world. The less sun shine in the Northern territories led to less melanin production. The black skin started getting more fair. Some legends and ancient drawings show that the Thracian Orpheus was a black man. Thousands of years later the Thracian Spartacus in the Roman Empire is a white man. Thracians were the first inhabitants of the present Bulgarian lands more than 7 000 years ago. They made the first worked gold in the world, the first script, the first grand buildings and are the basis of the development of the ancient civilizations. They have first selected Lactobacillus bulgaricus. proba.
|1. Stimulation of the immune system||They stimulate the production of B-lymphocytes which identify foreign microbes and production of phagocytes which destroy them. They increase the production of interferon which destroys viruses and the production of antibodies acting as antidotes. They increase the cytokinins binding the immune system as a whole;|
|2. Anti-tumor effect on the large intestine||Antimutagenic activity; they change the pro-carcinogenic activity of the colonized microbes and influence on the bile salts concentration;|
|3. Overgrowth in the small intestines||The lactobacilli influence on the activity of overgrowth, as they reduce the toxic metabolites and their anti-bacterial characteristics.|
|4. Heart diseases||Bacterial cells assimilate cholesterol; anti-oxidative effect; they change the hydrolase activity of enzymes.|
|5. Anti-hypertensive effect||Bacterial peptidases influence on the milk protein – a result from anti-hypertensive tripeptides; some components of the cell wall act as ACE inhibitors|
|6. Urogenital infections||Adhesion to the urinary and vaginal tract; they inhibit H202 production and restore the normal microbial flora|
|7. Infections, caused by Helicobacter Pylori||Competitive colonization, they suppress the development of Helicobacter Pylori.|
|8. Allergy||They protect from translocation of antigens in blood flow.|
|9. Resistance to inside pathogens||Resistance to colonizations; they create unfavourable condition for the pathogens (pH, bacteriocins)|
|10. Liver encephalopathy||They inhibit the urease-producing flora|
|11. They promote the lactose breakdown||The bacterial lactase influence on the lactose in the small intestines.|
Unlike other probiotic lactobacilli, during the lactic acid fermentation Lactobacillus bulgaricus produces D (-) lactic acid, which determines its powerful anticancerous, ray-protective and antioxidant effects. This acid together with the bacteriocins and other bioactive substances, secreted by Lactobacillus bulgaricus, detoxificate the organism.
Lactobacillus bulgaricus is the largest in size probiotic bacterium and through competitive relationship inhibits the development of pathogenic (disease-causing) microbes. Lactobacillus bulgaricus and Doderlein bacteria are part of the vaginal flora of women and determine her health and that of her children.
Studies of the Italian scientist Bianchi S. (1999) show that compared with the other probiotic bacteria, Lactobacillus bulgaricus adheres best to the mucous membrane of the large intestine and then rapidly reproduces itself in the body. This determines its more powerful healing and anti-toxic effect than the other probiotic bacteria.
Having enetered in the human body, Lactobacillus bulgaricus stimulates development of the other probiotic bacteria living there such as Lactobacillus acidophilus, Lactobacillus lactis, Bifidobacterium and others. Development of the other probiotic microorganisms is difficult and their beneficial effects are minimal without the help of Lactobacillus bulgaricus.
Development of the first probiotic products with Lactobacillus bulgaricus started by a team of scientists under the leadership of Retired Major General Prof. N. Alexandrov, D. Sc. in 1980 as a task set by the Bulgarian government for production of special foodstuffs for the international space program "Intercosmos". The produced dry probiotic foods are used by pilots from the Air Forces, Navy sailors, Bulgarian scientific expeditions to Antarctica and the Himalayas, also in cases of large-scale industrial accidents in Bulgaria. They were also used by seriously ill people and athletes training for Olympic and other world competitions.
The products became public in 1993 and this enabled their subsequent elaboration and improvement, as well as their production and supply on the open market.
The probiotic products with Lactobacillus bulgaricus developed by «Daflorn» Ltd. are unique without analogue in the world. They are the next step in technological respect as compared with the other probiotic products. Taken alone they are super concentrate of the original Bulgarian yoghurt, as only the beneficial ingredients are extracted from it biotechnologically and there is no need to consume the rest – about 95 % with no positive effects (even lactose has harmful effects on the cardio-vascular system). Comparison with kefir is the same. The probiotic bacteria contained in the Tibetan fungus of kefir are not enough. Kefir does not contain Lactobacillus bulgaricus and its durability is short – 2-7 days in refrigerator. The new formula of «Daflorn» Ltd. for biotech production mode allows 1 g dry substance of the product to contain billions of live and latent cells of Lactobacillus bulgaricus. During the biotechnological process the milk proteins and casein are broken down to all 21 essential and non-essential amino acids, the milk fats are broken down to polyunsaturated Omega 3 and 6 fatty acids, the milk sugars are broken down to simple sugars. They are ready for absorption by organism as structural units of the cells. During the production lactobacilli are naturally "enveloped" with gluco-protein coat, which allows them to remain alive at room temperature for months, even years. This process is subject to European patent protection and is the know-how of «Daflorn» Ltd.
We produce all our products as we keep good manufacturing practices and observe the requirements for Food Safety ISO 22000-2005. Our products are clinically tested in leading medical institutions in Bulgaria and abroad – MMA Sofia, Medical Universities in Varna, Sofia and Pleven, National Institute of Emergency "Pirogov", MMA in Czech Republic and Poland, Medical University of Würzburg - Germany, together with the Medical Control of the Soviet Army and the accredited doping laboratory in Cologne.
The products do not contain preservatives, genetically manipulated microorganisms, synthetic and chemical anabolic substances. They have no adverse effects. They can be taken in combination with each other and together with the basic medical treatment. Probiotics with Lactobacillus bulgaricus can be taken indefinitely and without interruption.
Our products are used by the Bulgarian troops during the missions in Cambodia, Kosovo, Iraq and Afghanistan, by people hurt by the earthquake in Turkey, by people working in heavy industry (cement production, asphalt bases, road construction, etc.) and by seriously ill people and children under treatment in hospitals.
Daflorn Ltd produces probiotic functional foods under the trade mark Biomilk in powder, wholefood supplements under the trade marks Biomilk, Laktera and Magic Power in capsules, cosmetic products and dry face packs Mariah's face and probiotic starter cultures (yeasts), containing Lactobacillus bulgaricus and dry herbal extracts and oils.
Special attention on the problem of physiological nutrition was paid in the beginning of XX century by I. P. Pavlov – Doctor Honoris Causa of over 120 scientific societies, academies and universities in Russia and all over the world. «The Father of the physiologists in the world" wrote that food is the ancient connection between people and the surrounding nature. According to Pavlov optimal nutrition must meet the enrgy and biological needs of organism.
Later on the concept of functional food was developed in Japan in the 80s of the last cantury. According to the Japanese Ministry of Education functional foods must have three basic functions: nutritive, sensory and healing.
European institutions have the following requirements to the functional foods:
- they must be taken every day;
- to be part of usual food;
- they must be obtained from natural products;
- to have a beneficial effect on human organism;
- to reduce risk of diseases and improve quality of life.
Widespread and not always rational use of medicines in medical practice leads to frequent allergies and toxic-allergic reactions. One of the serious complications of medical treatment is allergic-toxic epidermal necrosis or Layel syndrome. Almost always the disease occurs with severe infectious-inflammatory complications (pneumonia, sepsis), leading to increased break-up of the body tissues. Photos below show the beneficial results of treatment of a case with Layel syndrome, where to the main treatment was included food therapy with 200-400 g a day functional probiotic food Biomilk.
Our and foreign studies show that Lactobacillus bulgaricus alters the translocation of antigens and has a favourable effect on allergies. Studies by G. Neshev (1990) prove the beneficial effect of the Bulgarian rose oil in cases of anaphylactic shock, bronchial asthma, serotonin spasm and edema, induced experimentally in laboratory animals.
The combination of Lactobacillus bulgaricus and Bulgarian rose oil in the product Laktera Rose Oil makes it effective in allergic processes .
Lactobacillus bulgaricus in children
Children most often suffer from upper respiratory infections and diarrhea. The use of Bulgarian yoghurt with Lactobacillus bulgaricus for treatment of diarrhea in children in Bulgaria is a century-old tradition. In 1945 Dr. A. Fikov reported the successful treatment of infantile diarrhea with Bulgarian yoghurt, used for enemas in children's clinics in Berlin and Sofia. Later on G. Baudra (1990) and P. Solis (1993) published similar results for successful treatment of infantile diarrhea with Lactobacillus bulgaricus. Taking L. bulgaricus leads to increase of the mucins in the small intestines. The probiotic suppresses the development of viruses (most often rotaviruses) and removes them from the intestinal tract quickly. Lactobacillus bulgaricus produces a great amount of TNFa substance which stimulates the non-specific immune response. It also produces gamma interferon which blocks propagation of viruses. The probiotic overcomes disbacteriosis and regulates the gastrointestinal flora during and after antibiotic treatment.
These and other studies allow the World Health Organization in 1995 to recommend the use of Lactobacillus bulgaricus for treatment of infantile diarrhea.
During the clinical study in the children's clinic of the Medical University in Sofia with our probiotic products applied to children from 6 months to 1 year, diarrhea has not been observed and the rate of the respiratory infections have decreased by 50 %.
Lactobacillus bulgaricus and immune system
Immune system is the main protection of each individual. Conditionally it can be divided into two parts – local and general. More than 80 % of the immune cells are located in the intestinal lymph tissue. This is the «second brain» of organism, which aims to protect from the harmful aggression of pathogens. The general immune system is connected with blood, lymph, spleen, liver and bone marrow.
Immune system can be weakened when exposed to a large number of pathogens for a long time (eg. poor hygiene), in cases of physical and psychological strain, sharp temperature fluctuations, insufficient or poor nourishment, prolonged use of toxic or chemical substances – alcohol, drugs, medicines, in cases of prolonged contact with harmful chemical agents, radiation and others.
Scientific studies of H. Chomakov (2007), A. Hosono (2002), Link-Amster (1994), K. Thoreux (2001) and others show that Lactobacillus bulgaricus stimulates the immune system by increasing the production of:
- β lymphocytes which identify foreign bodies;
- phagocytes which destroy foreign bodies;
- YgA, YgG, YgM acting as antidotes;
- Interferon and cytokinins, supporting the fight against viruses.
In our clinical trials with the effect of Biomilk on Air Force pilots the obtained results are similar to those described above. Our studies show that distinct improvement of the immune resistance begins 7-10 days after the start of taking Biomilk and is at the maximum between the 15th and 20th day
Lactobacillus bulgaricus and gastrointestinal tract
The most common cause of gastritis, stomach and duodenal ulcer and gastric cancer is Helicobacter pylori. 50 to 70 % of the Europe's population is infected with it. Treatment with antibiotics for Helicobacter pylori is not successful because this bacterium develops an antibiotic resistance quickly and their toxic and adverse effects. Immediately after the antibiotic treatment, the organism is infected again which makes treatment useless. It was found out that Lactobacillus bulgaricus decreases the number of Helicobacter pylori in stomach on the basis of competitive elimination and by stimulation of the immune response it protects organism from infection.
Table 5 shows our experiment for treatment of ethyl alcohol induced ulcers in rats with different Laktera products containing Lactobacillus bulgaricus and herbal extracts.
|Test groups||Ulcer index (mm2)||Test groups||Ulcer index (mm2)|
|Marker||No ulcers||Ethyl alcohol 45o||24,3 ± 8,5 uclers|
|No ulcers||Ethyl alcohol 45o + Laktera optima||3 spot ulcers|
(probiotic + dry extract of hypericum perforatum)
|No ulcers||Ethyl alcohol 45o + Laktera relax||5 spot ulcers|
|Laktera forte (probiotic + dry extract of gin seng)||No ulcers||Ethyl alcohol 45o + Laktera forte||No ulcers|
It appears that Laktera Forte – probiotic containing Lactobacillus bulgaricus and gin seng – has the most powerful effect.
Bowel chronic inflammations (colitis, Crohn's disease, inflammations of ileum) affect 15-20 percent of the population in the developed countries and are the major health problem. Frequent use of antibiotics leads to development of serious disbacteriosis and makes favourable conditions for development of fungi in intestines. Recent studies show that the best results are obtained in treatment of intestinal diseases with lactobacilli combined with the main treatment. Figure 1 presents our experiment with 152 patients with gastroduodenitis, colitis and disbacteriosis for 6-month period of treatment only with Biomilk and Laktera.
Figure 1: Changes of clinical manifestations of the damages of the Gastrointestinal Tract for a 6-month period of treatment
The figure shows that after six months disbacteriosis is completely under control, colitis and gastroduodenitis have fallen down from 71 % to 4 %. The study shows that in cases of Crohn's disease, Lactobacillus bulgaricus must be taken for a long time – months and years and in the beginning it must be combined with the main treatment to obtain lasting beneficial effects. Without Lactobacillus bulgaricus, the effect of the traditional medical treatment is temporary and recurrences are common.
Lactobacillus bulgaricus and liver lesions of infectious and toxic origin
The main cause of toxic liver damage is alcohol. Figures 2 and 3 show the results from a complex treatment of addiction to alcohol and drugs in 152 patients. Food therapy with the probiotics Biomilk and Laktera was applied to all of them.
Figure 2: Changes in the parameters of ASAT and ALAT for a period of 6 months
Figure 3: Changes in the data from ultrasound diagnosis of liver for a period of 1 year
The figure shows that the values of liver enzymes and ultrasound image of liver are restored after six month.
Our clinical study confirms the experimental tests with electronic microscopy, conducted in the 80 years in the Bulgarian Academy of Sciences also that treatment with Lactobacillus bulgaricus leads to stabilization of the cell membrane and restores the functions of liver cell.
Other serious liver diseases are viral hepatitis. Our clinical studies in an epidemic of hepatitis A in 227 soldiers, divided into two groups: first - 123 patients with complex treatment received Biomilk Hepanorm Forte and Laktera Rose Oil and second - 104 patients with standard treatment of viral hepatitis, showed the following results:
- Treatment with Biomilk and Laktera was significantly more effective than the conventional treatment in the control group and shortens the duration of symptoms of the acute viral hepatitis;
- The probiotics Biomilk Hepanorm Forte and Laktera Rose Oil accelerate normalization of bilirubin, parameters of cholestasis and cytolysis and increase the amount of total protein.
Studies of number of scientists – A. Hosono (2002), H. Chomakov (2007) show that Lactobacillus bulgaricus has a direct anti-viral effect. It is determined both by strengthening of the immune system and by increasing the production of interferon and bacteriocins acting against viruses. Further the amino acids in the functional probiotic food Biomilk are assimilated by the liver as a building material for live and healthy cells.
This is confirmed by our study with 42 patients with HVC, treated without interferon, only with the probiotics Biomilk and Laktera.
Figure 4: Change of the values of the titre of antibodies against HVC in 42 patients treated with probiotic for a period of 1 year
The values of the titre of antibodies were measured in U / L, as the norm is up to 2 U/L. The figure shows that titre of antibodies against HVC in treatment only with Lactobacillus bulgaricus decreases significantly for a period of 1 year.
Lactobacillus bulgaricus and heart and cerebrovascular diseases
In the 20 century the death-rate from heart and cerebrovascular diseases increased many times. The main cause is atherosclerosis and high cholesterol. According to the general opinion nutrition and taking large amounts of saturated fatty acids and animal fats causes an increase in cholesterol, especially the bad cholesterol and triglycerides. Taking large amounts of these foods leads to increased excretion of bile acids through which cholesterol gets into intestines. In order to expel it from the body, it is necessary to combine with any of the following substances:
- unsaturated fatty acids;
- amino acids;
- lactic acid bacteria.
When the quantity of these components is insufficient, cholesterol remains free and is absorbed again by the human organism. This leads to its increase despite the implementation of different diets.
Table 6 presents our clinical study with 54 Air Force pilots from Bulgaria for 30 days, who did not change their daily regimen, their food was rich and they have taken the probiotics Biomilk and Laktera. .
Effect of the lactic acid probiotic product Biomilk on fat metabolism in military pilots (Table 6).
|Period||Total cholesterol||HDL cholesterol||LDL cholesterol||Triglycerides|
The table shows that cholesterol decreased by one unit for one month. The complex action of the probiotic Biomilk, containing Lactobacillus bulgaricus as well as unsaturated fatty acids, amino acids and calcium, is of great importance for the indicated beneficial results. The balanced nutritional composition of the probiotic Biomilk regulates metabolism of the human body completely.
Figure 5: Changes in metabolism for a period of 1 year
Our results obtained in 152 patients are presented in figure 5.
It is obvious that regular use of the probiotics Biomilk and Laktera in combination with the main treatment of alcohol dependence leads to control of consumative and bone articular syndrome and restoration of trophic of skin and hair for a period of about 1 year.
Anti-tumor and anti-mutagenic effect of the lactic acid bacteria
Factors that lead to cancer are three – carcinogens, mutagens and microorganisms. The process of cancer formation occurs in two stages – appearance of mutant cells as a result of damage of the cellular DNA and transformation of mutant cells into cancer cells. This process is shown in figure 6.
Figure 6 Process of cancer formation due to chemical agents
Factors leading to cancer exist in our life and we are constantly exposed to their harmful influence. Cancer cells are constantly formed in the human body, but phagocytes destroy them. When the immune system weakens, especially in combination with stress, the organism cannot respond adequately to the increased number of cancer cells and they begin to multiply uncontrollably.
Some studies, conducted in the Agricultural department of Shinshu University in Japan, show that lactic acid bacteria have anti-mutagenic effects. Table 7 shows the anti-mutagenic effects of different lactic acid bacteria and food against the mutagen AF2 .
|ANTIMUTAGENIC EFFECT OF DIFFERENT LACTIC ACID PRODUCTS (cultivated 24 hours) AGAINST AF2|
|Lactic acid products||Suppressive effect on AF2 (%)|
|0 hours||24 hours||increase|
|UHT milk – Lb. delbrueski subsp. Bulgaricus
UHT milk – St. Lactis subsp. Lactis
UHT milk – St. Salivarius subsp. Thermopillus
Skimmed milk – Lb. delbrueski subsp. Bulgaricus
Skimmed milk – St. Lactis subsp. Lactis
Skimmed milk – St. Salivarius subsp. Thermopillus
Whole milk – Lb. delbrueski subsp. Bulgaricus
Whole milk – St. Lactis subsp. Lactis
Whole milk – St. Salivarius subsp. Thermopillus
The information shows that within 24 hours AF2 loses its mutagenic effect to a great extent. Products, containing Lactobacillus bulgaricus, have the most beneficial anti-mutagenic effect at the end of the 24-hour period.
Based on these and other results, Hosono makes the following conclusions:
- The cell walls of the lactic acid bacteria (both live and dead) have anti-mutagenic and anti-carcinogenic effects;
- Lactic acid bacteria combine mutagens and carcinogens in insoluble complexes and they are removed from the body.
Similar results, demonstrating the anti-tumor effect of lactic acid bacteria, were obtained in experimental studies conducted by I. Bogdanov (1965) and later on by Kitadzava (Japan), Tsonevski and Enikova (1995). They found out that the anti-tumor effect of different lactobacilli is different. Lactobacillus bulgaricus, producing D(-) lactic acid has the best effect. The cancer rate in experimental animals, fed with lactobacilli, producing L(+) lactic acid, is higher.
Radio-protective and antioxidant effect of Lactobacillus bulgaricus
The influence of the ionizing radiation is not only a military threat. Widespread use of nuclear energy, radiotherapy, X-rays requires creation of radio-protective remedies. Most of them are chemicals with transient effect. They have many adverse and toxic effects, which are exalted in cases of taking them for a long time, which is necessary for people working in nuclear energetics and with isotopes and in cases of radiotherapy.
Experimental studies of Z. Mitsov (1960), B. Gyosheva, Z. Goranova (1992) show that Bulgarian yoghurt with Lactobacillus bulgaricus decreases the initial damages of bone marrow, accelerates restoration of haematopoiesis and decreases the death rate of experimental animals, irradiated with radioactive agents, in comparison with the control group.
Experimental studies with Biomilk conducted by B. Gyosheva (2005), M. Georgieva (2007) show that the experimental animals that have taken Biomilk, normalize haematopoiesis of the bone marrow faster on the 10th day after irradiation.
The results are shown in figure 7.
Most likely the ray-protective effect of the probiotic Biomilk is due to its immuno-stimulating effect and to limiting of the lipid peroxidation, i.e. of free radicals. There are other studies by B. Bay (1999), Lin My (1991) and others on the antioxidant effect of probiotic bacteria.
The study of A. Gigov – Department of Hematology and Oncology at the Military Medical Academy Sofia 2008 – with 95 cancer patients deserves special attention. The patients have taken Biomilk Antioxidant in combination with the main treatment and there was statistically significant increase in the number of T-lymphocytes, thrombocytes and leukocytes and a moderate increase of hemoglobin. They have taken chemo- and radiotherapy very well and their quality of life has improved. Similar results were obtained in 1976 by A. Atanasova in 36 children with leukemia and by us in 70 patients with low thrombocyte counts due to prolonged treatment with corticosteroids.
Development of bio radio-protectors is essential for workers in nuclear power, cancer patients undergoing chemo- and radiotherapy.
These studies and facts are part of 30 years work. Now and in future we will try to connect the knowledge of the ancient civilizations with modern scientific achievements and to restore the message of Hippocrates:
"Let thy food be thy medicine"