Lactobacillus Bulgaricus - Gift from the gods

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 species;
  • beneficial (probiotics) – about 15-20 species;
  • latent (conditionally pathogenic) – over 1000-1200 species;

Probiotics are live microorganisms which administered alone or via foods that contains them, 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)

Lactobacilli

Lactic acid cocci

Bifidobacteria

Lactobacillus bulgaricus Streptococcus thermophilus Bifidobacterium bifidum
Lactobacillus acidophilus Lactococcus faecium Bifidobacterium longum
Lactobacillus casei Lactococcus lactis Bifidobacterium infantis
Lactobacillus plantarum Bifidobacterium breve
Lactobacillus rhamnosus
Lactobacillus reuteri
Lactobacillus helveticus

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 invention of 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 occurred first in Thracians and then in the other Europeans. The new lactic acid food and the billions of beneficial (probiotic) bacteria created a gene, coding the production of the enzyme lactase which breaks down milk sugar in human body. This new technology granted the rare privilege people to be able to consume dairy products obtained from the milk of the domestic animals throughout their entire lives. People began obtaining the vital vit. D from the lactic acid products, and not only from the sun, as it was till that moment. 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 processed gold in the world, the first script, the first grand buildings and are the basis of the development of the ancient civilizations. They are the first people, who cultivated Lactobacillus bulgaricus probiotic.

Lactobacillus bulgaricus is of vegetable origin and is isolated from green plants only on the territory of Bulgaria. In other geographic regions of the world it mutates and is no longer able to reproduce. That’s why countries as China, Japan, Korea, Italy and others, offering products containing Lactobacillus bulgaricus, buy it from Bulgaria. Based on summarized data from leading scientists from USA, Japan, France (G. Tannock, A. Hosono, G. Complien) and according to our information Lactobacillus bulgaricus has beneficial effects in 11 directions. They are shown in Table 2.

Beneficial effects of probiotics (Table 2)

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.

Lactobacillus Bulgaricus - the largest probiotic bacteria
Bifidobacterium

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 my leadership (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.

The functional probiotic food Biomilk is a new generation entirely balanced functional food. It combines the healing effect of Lactobacillus bulgaricus and the nutritive and energy effects of the nutrients broken down during the lactic acid fermentation.

Table 3 shows amino acids and their amount in 100 g Biomilk Power Energy. The correlation between essential and non-essential amino acids is from 1:2,2 to 1:2,4, as the physiological rate is 1:2,8. With these parameters the functional food Biomilk Power Energy meets the international requirements for a standard protein food.

Different amino acids and their content in 100 g Biomilk (Table 3)

Amino acids
Content in 100 g product
Amino acids
Content in 100 g product
Essential
Non-essential
1. Isoleucine
4.81-5.02
11. Alanine
3.27-3.30
2. Leusine
9.02-9.16
12. Arginine
3.15-3.28
3. Lysine
5.71-6.02
13. Aspartic acid
7.06-7.42
4. Methionine
3.06-3.08
14. Glutamatic acid
19.75-20.65
5. Phenylalanine
4.98-5.48
15. Glycerine
5.52-5.81
6. Threonine
4.25-4.28
16. Histidine
3.55-4.24
7. Tyrosine
1.92-3.12
17. Proline
9.80-10.02
8. Valine
5.86-6.07
18. Serine
5.44-5.91
9. Tryptophan
0.63-0.64
19. Lisinoalanine
0.20-0.26
10. Cystine
0.48-0.53

Global Scientific Researches from 2010 show that the amino acids leucine, isoleucine and valine prolong human life. They are called “Cocktail of life.”

Biomilk Power Energy contains an appropriate quantity of Omega 3 and Omega 6 low molecular capronic, caprylic, capric and lauric fatty acids, as well as the essential linoleic and linolenic fatty acids. The balanced proportions between the different fatty acids in Biomilk make it an important product for prevention of atherosclerosis that promotes the synthesis of different hormones and enzymes.

Different fatty acids and their content in 100 g Biomilk (Table 4)

Fatty acid
% content
Fatty acid
% content
C4 : 0
1.0
C16 : 0
1.8
C6 : 0
0.5
C16 : 0
1.5
C8 : 0
1.1
C16 izo
C10 : 0
2.7
C17 : 0
1.0
C12 : 0
3.6
C18 : 0
9.6
C12 : 0
C18 : 0
23.9
C14 : 0
11.8
C18 : 0
3.1
C14 : 0
1.1
C18 : 0
0.08
C14 : 0
0.7
C20 : 0
1.0
C15 : 0
1.6
C20 : 0
0.1
C15 : 0
0.4
C22 : 0
0.1
C16 : 0
33.0
Others
1.4

Carbohydrates in Biomilk Power Energy are broken down biotechnologically to easy of digestion monosaccharides – glucose and galactose. The products are low lactose and can be taken by people lacking in lactase enzyme (intolerant of milk).

Atom-absorbing analyses shows that Biomilk contains great quantities of all basic macro-and microelements – calcium, potassium, phosphorus, magnesium, iron, zinc and others. It also contains number of essential vitamins A, B1, B2. Increased amount of vit. D3 is important for the rapid and full digestion of the high calcium content in Biomilk. This determines the apparent effect of the products in cases of osteoporosis and low sunshine in winter and in northern regions. All nutrients in Biomilk are ready for absorption by the human organism. People do not consume energy for this and increase their energy supply for brain activity and substitution and rejuvenation of the cells.

The functional probiotic food Biomilk is suitable for children, people put to severe physical or mental work, people exposed to stress and strain and the harmful influence of the environment. Biomilk improves and prolongs life.

Herbal extracts and oils with proven healing effects are added to some of the products. The amino acids in the products serve as a quick transport carriers and help the full absorption of the extracts and oils by the body cells, as together with Lactobacillus bulgaricus they exalt their beneficial effects many times. This makes our products materially different from the other probiotic products and makes them a leader among modern probiotics.

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 allergy

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 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.

Table 5

Test groups Ulcer index (mm2) Test groups Ulcer index (mm2)
Marker No ulcers Ethyl alcohol 45o 24,3 ± 8,5 uclers
Laktera optima
(pure probiotic)
No ulcers Ethyl alcohol 45o + Laktera optima 3 spot ulcers
Laktera relax
(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

Figure 1
Figure 2
Figure 3
Figure 4

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

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.

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;
  • calcium;
  • fibres;
  • pectin;
  • 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
1st day 7.10 1.23 4.32 3.68
30th day 6.05 1.49 3.85 3.57
60th day 5.02 1.51 3.52 1.72

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

Figure 5
Figure 6

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 .

Table 7

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
68.5
71.6
66.7
65.8
70.2
70.1
66.2
71.0
63.0
80.2
80.8
74.3
79.1
81.8
83.2
82.5
71.3
71.0
11.7
9.2
7.6
13.9
11.6
13.2
16.3
0.3
8

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”


Nutritional therapy with probiotic Laktera in COVID 19 patients

Nutritional therapy with probiotic Laktera, containing strain Lactobacillus bulgaricus DWT1 from water origin in COVID 19 patients

Unique Bulgarian probiotic strain Lactobacillus bulgaricus DWT1 from water origin is used in treatment of COVID 19 patients. For the first time Laktera probiotic is used in patient of the University hospital, Pleven. The patient in questioned is a 56 year old man, treated for 30 days with pneumonia, resulting from COVID-19. Standard treatment was applied with quadruple antibiotic combination, tonics and corticosteroids. The patient was discharged from the hospital with improvement, but a few days later was admitted again in severe condition with rash, which affected the entire body and mucosa. The rash was bleeding when touched. The mouth mucosa was also severely affected and the person was unable to eat. A case with such clinical picture and type of rash is described for the first time in Bulgaria. There are scientific data from Italian and English physicians, describing similar rashes in COVID 19 cases. Most probably this rash is due to affection of the small blood vessels of skin and mucosa. The patient begin afresh treatment with triple antibiotic combination, protein solutions and tonics, but with no improvement. On the third day of this treatment, to the overall treatment regimen is included probiotic Laktera, containing Lactobacillus bulgaricus DWT1 from water origin in dosage of 15 grams, taken two times a day.  On the second day of probiotic Laktera administration, the rash began to disappear and the skin to recover. The patient began eating, due to mucosal restoration. On the tenth day, the patient was signed out from hospital with healthy skin and mucosa, without fever and with negative PCR test for COVID-19.

In the same University hospital in Pleven are admitted 31 patients with positive PCR test for COVID-19. For them the disease is manifested with breathing difficulties, high temperature and roentgen graphic data for bilateral pneumonia. From the first day after the hospital admission, all these patients, besides the main treatment are taking nutritional therapy with probiotic Laktera in dosage of 15 grams, taken two times a day. It is notable that unlike other patients with pneumonia, caused by COVID 19, the ones, who take Lactobacillus bulgaricus DWT1 from water origin improve much faster. On the third-fourth day of the hospital treatment, they are in good general condition, with no fever or manifestations of breathing difficulties and very reduced caught. On the ninth-tenth day after the hospital admission, PCR for COVID-19 is negative and the pneumonia is overcome. There are no diseased COVID-19 patients, who are taking probiotic Laktera. The clinical observation and application of nutritional therapy with Laktera in COVID-19 patients continues even at the moment.

We also apply nutritional therapy with Laktera probiotic in patients with milder forms of COVID 19, treated at home. They appear to recover much faster than other COVID-19 patients also treated at home. The patients, who have taken Laktera overcome the usual symptoms of COVID 19 such as tiredness, lethargy and lack of concentration in a matter of three-four days. No one of the so far observed 5 ambulatory treated COVID 19 patients, who have taken Lactobacillus bulgaricus DWT1 from water origin did not need hospital admission due to complications.

The action mechanism of Lactobacillus bulgaricus DWT1 onto COVID-19 was discovered by prof. Palok Aich of National Institute of science Education and Research of Department of Atomic Energy, Government of India. Two years ago he proved that this strain of water origin converts M2 to M1 macrophage cells. The M1 macrophages are our own immunity cells, produced by the human organism. The M1 macrophage have strong stimulating effect onto the cell immunity and kill tumor and virus cells. You can watch the full lecture of prof. Aich on the subject here. 

The Laktera probiotic is produced by the Bulgarian company Daflorn Ltd and is certified by the Bulgarian Food Safety Agency as special purpose food. The strain Lactobacillus bulgaricus DWT1 is the first strain in the world, isolated from pure spring water in Bulgaria and the probiotic is patented in the USA with Patent № US 9,131,708 B2 and Patent in the European Union №3172973.

prof. Dr. Nikola Alexandrov MD DSc.

Dr. Daniela Petrova MD PhD

Dr. Georgy Aleksandrov PhD

Maria Peneva PhD

Date: 20.07.2020

Sofia

Bulgaria


The invention of Daflorn

The new discovery of Daflorn is isolation of a probiotic association of lactic acid microorganisms from spring water in the wild nature in Bulgaria – a spring in the Balkan mountains.

The probiotic association contains 8 wild all natural strains of 4 species lactic acid bacteria of Lactobacillus bulgaricus, Lactobacillus helveticus, Lactobacillus lactis and Streptococcus thermophilus. The original strains of the probiotic association live in spring water.

Isolation of probiotic strains from a natural water source raises the question of revision of the popular belief in the scientific community, that the probiotic lactic acid bacteria can be isolated only from dairy products, plants, animals and man. Capture of probiotic microorganisms from spring water happens for the first time in the world, so we can talk about a scientific discovery. That discovery gives a great advantage and sky-rocks Daflorn to an indisputable leader in innovation in the production of probiotic products.

We managed to ferment typically pasteurized milk from the commercial network with spring water.

This is a unique method for capturing of wild all natural probiotic microorganisms from spring water. The lactic acid microorganisms in the probiotic association live and develop in natural aquatic environment in the constantly changing climatic conditions in the wild.

Our new wild probiotic microorganisms are registered under the Budapest Treaty on the International Recognition of deposit of microorganisms for the Purposes of Patent Procedure in the Czech bank for microorganisms.

  • strain Lactobacillus delbrueckii subsp. bulgaricus DWT1, registered at CCM under No. 7992
  • strain Lactobacillus helveticus DWT2, registered at CCM under No. 7993
  • strain Lactobacillus delbrueckii subsp. lactis DWT3, registered at CCM under No. 7994
  • strain Streptococcus thermophilus DWT4 registered at CCM under Number 7992
  • strain Streptococcus thermophilus DWT5 registered at CCM under Number 7993
  • strain Streptococcus thermophilus DWT6 registered at CCM under Number 7994
  • strain Streptococcus thermophilus DWT7 registered at CCM under Number 7995
  • strain Streptococcus thermophilus DWT8 registered at CCM under Number 7996

The microorganisms in the probiotic association, isolated from spring water, are unique and differ from the probiotic microorganisms, found in the plants (grass, flowers, plants and algae) in and around the water.

The new probiotic microorganisms, method of obtaining the products and the products are subject to patent protection of Daflorn, application number 111161/07.03.2012 in Bulgaria and Patent number US 9,131,708, B2 in Patent office in USA..

Daflorn Ltd uses these all natural probiotic strains for the production of unique probiotic products. During the biotechnological production mode the lactic acid microorganisms are naturally enveloped with the amino acids, fatty acids, simple sugars, micro elements, minerals and vitamins of milk, obtained as a result of fermentation. During the biotechnological production as approaching the fermentation process as close as possible to the natural one, Daflorn managed to create the most ancient and at the same time the most modern generation all natural probiotics.

Until now there are 4 known generations probiotics in the probiotic production:

  1. 1st Generation – non coated probiotics
  2. 2nd Generation – enteric-coated probiotics
  3. 3rd Generation – micro ecapsulated probiotics
  4. 4th Generation – dual coated probiotics – ph dependent release system , moisture, temperature and pressure protection

All of the above generations are scientifically manipulated by man.

X generation of Daflorn is the oldest and at the same time the most modern generation of all natural probiotics

Advantage of the new generation of Daflorn

We put the nature in the science, in the biotechnology and during the biotechnological mode of production of Daflorn the probiotic microorganisms naturally “envelope” themselves with natural coat, consisting of lactic amino acids, fats, vitamins and minerals, which preserves them alive at a room temperature (20° -24° C) for years.

Despite the high sensitivity of Lactobacillus bulgaricus, Lactobacillus helveticus, Lactobacillus lactis and less of Streptococcus thermophilus to lyophilization (freeze drying at -45° C under vacuum for 48 hours) the probiotic microorganisms in the association survive during this way of freeze-drying and stay alive in the obtained dry probiotic products. The probiotic bacteria from the association survive after rehydration when they pass through the gastrointestinal tract. The study was conducted under extreme conditions – pH 2.5 for 2 hours and after that at pH 8 for 5 hours. The wild probiotic strains have excellent viability and can optimally colonize the intestines. Their maximum activity of probiotic bacteria manifests there and that ensures their high efficiency.

Survival and stability of the probiotic bacteria from the association is due to the natural coat with which they are enveloped during the biotechnological process as well as to their natural water origin. In nature they have adapted to the changing climatic conditions and have acquired very high viability. That natural coat and the water origin of the probiotic microorganisms ensure them the stability and maximum bioavailability.

In the probiotic association the strains of lactic acid microorganisms multiply and develop together which is a new step in science. Until now it was known that Lactobacillus bulgaricus develops only together with Streptococcus thermophilus. In the new X generation found by Daflorn the 8 strains of 4 types lactic acid microorganisms Lactobacillus bulgaricus, Lactobacillus lactis, Lactobacillus helveticus and Streptococcus thermophilus live and multiply together. This enables the obtaining of products, extremely rich in live all natural probiotic microorganisms, and thus they are extremely valuable for healthful nutrition of man.

Lactobacillus bulgaricus and Lactobacillus helveticus are probiotic microorganisms that carry out homo-fermentation. During that fermentation pure lactic acid is produces and bacteriocins (biologically active substances) are released. They have a beneficial effect on the human body. During reproduction Streptococcus thermophilus produces the enzyme alcohol dehydrogenase. That enzyme helps the faster breakdown of ethyl alcohol and recovery of the body after alcohol intake.

The products containing the probiotic microorganisms from the association – Lactobacillus bulgaricus, Lactobacillus helveticus, Lactobacillus lactis and Streptococcus thermophilus have a good antimicrobial activity against harmful (pathogenic) microorganisms. They have a moderate bactericidal effect – kill pathogenic strains of Staphylococcus aureus, which causes purulent inflammatory processes. Probiotic microorganisms exhibit a strong bacteriostatic effect – stop the growth of harmful strains of Escherichia coli, Salmonella, Shigella flexneri, Proteus mirabilis and Staphylococcus aureus, which cause severe disorders of the gastro-intestinal tract and other human organs – diarrhea, disbacteriosis and inflammations.

The probiotic bacteria from the association increase human body resistance to harmful microorganisms, which determines their health effects.

The probiotic products of Daflorn contain in 1 g over 1 x 108 live cells of the all natural strains, isolated from spring water – Lactobacillus bulgaricus, Lactobacillus helveticus, Lactobacillus Lactis and Streptococcus thermophilus. Their amount is enough and it determines their health benefits.

Main advantages of the probiotic association of Daflorn

  • Completely natural, isolated from spring water, unique probiotic strains
  • Maximal high survival and resistance during the production process
  • Maximal high survival and resistance during storage at a room temperature
  • Maximal high survival and resistance during passing through the gastrointestinal tract
  • Maximum bioavailability
  • Maximum beneficial effect against pathogens

Nutritional agents in the products

The products of Daflorn contain not only probiotic microorganisms but also nutrients. During the biotechnological process with the help of probiotic microorganisms from the association milk proteins and casein break down to all 21 essential and non-essential amino acids – Cystine, Isoleucine, Leucine, Lysine, Methionine, Phenylalanine, Threonine, Tryptophan, Tyrosine, Valine, Alanine, Arginine, Aspartic Acid, Glumatic Acid, Glicine, Astidine, Histidine, Hydr. Proline, Proline, Serine, Lisinoalanine. Milk fats break down to polyunsaturated milk acids. During that process probiotic microorganisms and in particular Lactobacillus bulgaricus breaks down lactose. Lactic acid bacteria from the association break down the long chains of nutrients, contained in milk during fermentation to nutritional components ready for absorption. By taking these nutrients the human system does not use up energy and thus spares the organs.

Amino acids

The products contain not only the non-essential but also all essential amino acids that the human body cannot produce as thus they should be imported by food ready to be absorbed. The proportions of the essential and non-essential amino acids is from 1:2,2 to 1:2,4. The probiotic product with these parameters meets the recommended international requirements for a standard protein food product. Amino acids in the product are ready for absorption by the organism as building units of the cells and thus people do not overload the liver and pancreas to break down the proteins.

Probiotic products contain the most important muscle building amino acids – Leucine and Arginine. In the products they are ready to be directly absorbed and to build muscle filaments. The products contain also Lysine, Methionine and Tryptophan in large amounts. These amino acids are essential and are of animal origin. They participate in the breakdown of the other proteins of plant and animal origin (phytoproteins, casein, whey protein). Without them the breakdown of the other types of proteins is incomplete, they cannot be absorbed by the system and cause putrefactive processes in the large intestine and intoxication of the body. The product also contains the so called „Cocktail of life“ – the amino acids Leucine, Isoleucine and Valine, which prolong life according to Italian scientific research from 2010.

Fatty acids

The probiotic products contain the main types saturated and unsaturated fatty acids. They contain an appropriate for the body amount of low molecular capric, caprylic, caprynic and lauric acids that have an expressed anti-atherosclerotic effect. They also contain the essential lipoic and linolenic acids which the body cannot produce alone and they must be imported from outside by food. The products contain the optimum and balanced amount of fatty acids with a carbon number from 6 to 12 and from them the human body produces steroids and sex hormones which are very important. The products of Daflorn do not contain cholesterol. The company also offers low fat and defatted products.

Carbohydrates

In the probiotic products of Daflorn carbohydrates are broken down biotechnologically to easy for absorption monosaccharides – glucose and galactose as during production some of them are partially caramelized and give a nice natural caramel taste to the product. They easily reach and are instantly absorbed by the brain and liver. During fermentation Lactobacillus bulgaricus breaks down lactose because it feeds on it. The products are low lactose to lactose-free and are suitable for people with lactose intolerance. The products do not contain gluten and are suitable for people with gluten intolerance.

Vitamins and minerals

The probiotic products contain also the basic minerals – calcium, potassium, phosphorus, iron, etc. They are components of milk in the form of soluble compounds as they are fully absorbed by the human body. Calcium has a structural role in the human body as it is an essential element in bones and teeth building. Phosphorus participates in calcium metabolism and plays a principal role in its absorption. Iron is an essential element necessary for the hemoglobin production. Lack of iron can lead to reduced hemoglobin in blood and hence anaemia and fatigue.

The products also contain a number of milk vitamins which are of a great importance for the organism – Vitamin A, Vitamin B1, B2, Carotene. Vitamin A (retinol) is necessary to the eye retina to improve vision in low-light and for colour vision. It contributes to formation of growth factor. Carotene is a precursor of Vitamin A, i.e. Vitamin A is generated from it. Vitamin B1 (Thiamine) stimulates production of energy from glucose. Lack of that vitamin leads to neurological dysfunctions. Vitamin B2 (Riboflavin) participates in oxidative reactions of the respiratory chain. The content of vitamin D3 is essential because it is very important for the full absorption of calcium by the human body. Vitamin D3 is essential for people who live in northern areas and regions with less sunshine.

The combination of micro elements, minerals and vitamins from group B and A, which are actively involved in many essential metabolic processes, leads to balancing the basic metabolic processes in the human body.

The probiotic products of Daflorn are completely natural as they do not contain preservatives, colouring agents, synthetic and harmful substances and genetically manipulated microorganisms.

  • they do not contain gluten
  • they do not contain cholesterol
  • they are lactose-free to low-lactose

We add coffee, herbal extracts, oils, fibres, enzymes, honey to some of our probiotic products. The amino acids of the probiotics in these new products serve as a fast transport carriers as they catch the molecules of the added extracts and carry them quickly to the cells. Thus they potentiate the faster and full effect of the added coffee, herbal extracts and honey. By the unique combination of wild all natural probiotic microorganisms, nutrients, coffee, herbal extracts and honey the products of Daflorn endow body and mind with greater welfare.


Food allergy

Food allergy

Epidemiological study of patients with food allergies

M. Peneva, MC „Evrozdrave-Bulgaria”, excerpt from Abstract of dissertation, Sofia 2016, p. 27-29

Food allergies have significant harmful effects on families, economics, social interactions, school, work and service, as well as on quality of life related to health.

According to the medical observations, people with higher living standard and more active immune prophylaxis are more often affected by allergies.

The incidence of allergic diseases is increasing among all age groups. One of the most common allergies is the food allergy. For example, allergy to peanuts has doubled over the last decade. Food allergy is defined as an immunologically mediated hypersensitivity reaction to food proteins that are harmless, but alien to the human immune system. Major risk factors for the emergence and development of food allergy are the family medical history and environmental factors, especially important of which is the intake of foreign proteins such as cow’s milk, eggs, fish, soy.

A clinical study of food allergies involving 58 patients aged between 3 and 81 years was conducted in MC Evrozdrave in the period 2009 – 2014 in cooperation with specialist physicians. 6 of the patients were children aged from 3 to 9 years. 29 patients were women aged from 22 to 81 years and 23 were men aged from 24 to 71 years. All patients had different manifestations of allergic reactions. In some of them we observed classic symptoms of allergy – lacrimation, rhinitis, various rashes, itchy skin, diarrhea. In some patients we observed newly found symptoms of food allergy – hair loss, dandruff, periodontal disease, elevated levels of prolactin in young women. All patients have undergone immunoassay tests for 19 groups of food allergens by the ELISA method – immunomodulation for determination of the number of the specific antibodies against certain food allergens in blood serum. The patients were submitted to allergy tests to 19 groups of food proteins. All patients were found to be hypersensitive towards more than one food allergen.

The high percentage of food allergy to peanuts, seafood, strawberry, egg whites, cocoa and cow’s milk should be noted. Only one patient was found to be allergic to goat’s milk. Most likely this is due to the fact that the composition and nutritional functions of goat’s milk are the closest to the breast milk. A difference should be made between allergy to milk proteins and insufficient production of the enzyme lactase in the body, which enzyme breaks down lactose (milk sugar). After establishing the type of food allergy, we advised all patients to stop the consumption of food containing the respective food allergens, we recommended antiallergenic treatment with claritine 1 to 2 tablets daily depending on the severity of the allergy and probiotic treatment with Laktera Nature 15 g daily dose in combination with Laktera Rose oil 2 capsules a day. The probiotic dietary therapy for the patients with an established allergy to cow’s milk protein was a probiotic with goat’s milk Laktera Allergy free 15 g daily dose in combination with Laktera Allergy Free Rose+ – 2 capsules a day. Probiotics containing Lactobacillus bulgaricus combined with Bulgarian rose oil significantly reduce allergy symptoms and the allergic predisposition of the body.

Additionally, but not related to the allergic process, Lactobacillus bulgaricus improves lactose digestion and it has an approved health claim pursuant to Regulation 432/2012 of the European Union. Clinical symptoms of food allergy have faded away for a period of 2 to 6 weeks depending on the severity of allergy. We explained to the patients that they have to select and purchase food products carefully by checking the information on the labels about the composition of the food and the allergens contained therein. We advised the patients to actively look to allergens in the menus of the public restaurants.

The results of the study show that allergic diseases have increased considerably over the last decade. Food allergy can now be referred to the socially significant diseases. Obviously, food allergy is widely spread among the working-age population and necessitates specialized medical care. It requires a complex therapy with medical specialists, health managers and experts on food safety. Food allergy causes great social, economic and psychological damages to the population. People suffering from food allergy should be concerned with their health keeping anti-risk behavior and healthy lifestyle by active and targeted search of allergens in products before consumption.


European Health Claims

Lactobacillus Bulgaricus statement

LBB_Statement

Magnesium statement

Magnesium_statement

Magnesium statement 2

Magnesium_statement2

Oli Ola statement

Oli_Ola_statement

Anticancer activity of probiotics – experimental and clinical studies

Anticancer activity of probiotics – experimental and clinical studies

Georgi Alexandrov1, Kaloyan Georgiev2, Marieta Georgieva2, Ivan Iliev3, Maria Peneva4

Abstract

Introduction: Many scientists have been working on the anticancer effect of probiotics for the past decades. In the middle of the last century I. Bogdanov proved experimentally that Lactobacillus subsp. bulgaricus inhibits the growth and proliferation of tumor cells of Sarcoma 180, implanted in rats. Kitadzava (Japan) ascertained good anticancer effect of fermented milk on rats with implanted tumor cells. A. Hosono (Japan) found out that anticancer effect of fermented milk is due to the lactic acid bacteria and polysaccharides contained in it. Our preliminary studies showed that probiotic Laktera Nature® has an anti-proliferative effect against aggressive colon carcinoma cell line HT-29. The probiotic Laktera Nature® contains original strains of Lactobacillus bulgaricus DWT1 and Streptococcus thermophilus DWT4, isolated from spring water in Bulgaria and 21 natural essential and non-essential lactic amino acids. The aim of this study is to determine the anticancer activity of the different ingredients of probiotic Laktera Nature® and to study the effect of the probiotic formula as a nutritional therapy in patients with colorectal cancer.

Materials and Methods: Four types of cell lines have been used: 3T3, BJ, HT-29 & MDA-MB-231. Cytotoxicity was determined by MTT-dye reduction assay. The direct cytotoxicity of the tested cell lines, measured at the 24th hour and the antiproliferative activity, measured at the 72nd hour have been studied. Four different samples of Laktera Nature® were used: sample 1 (original probiotic formula containing live cells of Lactobacillus bulgaricus DWT1 and Streptococcus thermophilus DWT4 and 21 lactic amino acids), sample 2 (only the probiotic strains without the amino acids), sample 3 (only the 21 lactic amino acids without the probiotic strains) and sample 4 (the original probiotic formula that has passed γ-sterilization 10 Gy, whereby the probiotic strains are killed, leaving only the cell walls). The clinical study is carried out on 13 patients with surgery for colorectal cancer and undergoing chemo- and radiotherapy. Half of the patients take 15 g daily dose Laktera Nature® (sample 1) as a nutritional therapy.

Results and Discussion: The four samples show a negligible direct cytotoxicity of the tested cell lines measured at the 24th hour. Samples 3 and 4 show a slight concentration- and time-dependent cytotoxic effect. It was interesting to observe that sample 2 (only the probiotic strains) did not have the expected inhibitory effect on carcinoma growth (in that case MDA-MB-231), it even increased its growth. Sample 1 (the original probiotic formula Laktera Nature®, containing live probiotic strains and 21 amino acids) demonstrated a significant growth inhibition of cancerous HT-29 and MDA-MB-231 cell lines with IC50 values – 711,69 ± 82,1 and 934,21 ± 43,5 mcg/ml respectively, measured at the 72nd hour. The parallel conducted clinical trial shows that the nutritional therapy with the original probiotic formula Laktera Nature® allows the full courses of treatment with chemo- and radiotherapy to be carried out and reduces the side effects of the chemo- and radiotherapy.

Conclusion: The original probiotic formula Laktera Nature® inhibits growth of cancerous HT-29 and MDA-MB-231 cell lines. The nutritional therapy with Laktera Nature® improves quality of life and increases the survival rate of patients with colorectal cancer.

Keywords: Probiotic, Laktera Nature®, Lactobacillus bulgaricus, amino acids, antitumor effect, anticancer effect, cytotoxicity, nutritional therapy, colorectal cancer.

1 – Military Medical Academy – Sofia, Bulgaria

2 – Department of Preclinical and Clinical Sciences, Sector Pharmacology and Toxicology, Faculty of Pharmacy, Medical University Varna, Bulgaria

3 – Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences, Sofia, Bulgaria

4 – Medical Centre “Evrozdrave” Sofia, Bulgaria

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Meta-analysis for the effects of probiotic product Laktera with Lactobacillus bulgaricus on gastrointestinal tract

Meta-analysis for the effects of probiotic product Laktera with Lactobacillus bulgaricus on gastrointestinal tract

Summary

The probiotic product Laktera with Lactobacillus bulgaricus regulates and recovers the functions of the gastrointestinal tract and heals the internal human ecosystem.

The aim of Konstantin Ramshev and George Alexandrov from the Military Medical Academy in Sofia and Marieta Georgieva from the Medical University – Varna, is to prove the beneficial effects of a probiotic product Laktera with Lactobacillus bulgaricus on the human gastrointestinal tract.

Researchers have reviewed medical data published in Bulgarian and English languages in twenty two reports, exposés, dissertations and scientific articles for randomized controlled clinical studies on 2550 adult patients and therapeutic naïve individuals. These clinical studies were carried out within 31 years in Military Medical Academy, Medical Universities, Scientific Research Laboratories and Institutes in Member States of the European Union. The clinical studies were preceded by experimental trials on experimental animals. People, subject of these clinical studies were observed for adverse and toxic effects of the probiotic product for a subsequent period of over 10 years and such have not been registered.

The analysis includes clinical trials on humans for evaluation of the probiotic product Laktera with Lactobacillus bulgaricus as a part of tube and/or enteral feeding or part of a complex treatment regimen for sick people with various diseases and severe multiple organ damages and coma,  for healthy people put to extreme physical and mental stress as well as pilots from the Air Force, military sailors and astronauts.

Results

Regular intake of the probiotic product Laktera with Lactobacillus bulgaricus as a part of a complex treatment and enteral feeding leads to a significant reduction of the damage of the gastrointestinal tract, described below with substantial statistical significance. The probiotic products Laktera with Lactobacillus bulgaricus are efficient in cases of the following damage of the functions of the gastrointestinal tract:

  • Disorder of the functions of the gastrointestinal tract in case of space sickness of movement and extreme stress
  • Disbacteriosis and disorders of the intestinal flora of different etiology
  • Gastroenteritis and colitis of toxic, viral and allergic origin
  • Impossibility of active food intake and hypoproteinaemia in comatose, early postoperative period, severe chemical burns and toxo-allergic damages (Lyell syndrome) of the gastrointestinal tract.
  • Impaired digestion
  • Upper dyspeptic syndrome
  • Irritable Bowel Syndrome
  • Increased flatulence
  • Consumptive syndrome and hypoproteinaemia in late postoperative period and due to drug addiction
  • Gastroenteritis and diarrhea as a result of infectious inflammatory damage to the gastrointestinal tract caused by Salmonella, Shigella, coliforms, Proteus and staphylococci
  • Haematemesis of toxic origin (ethyl alcohol)
  • Gastro-enteropathy as a result of lactase deficiency and lactose intolerance
  • Gastro-enteropathy as a result of beta-galactosidase deficiency and galactose intolerance (galactosemia)
  • Disorder of the intestinal motorics
  • Ulcers and gastritis of toxic, toxo-allergic, corrosive or infectious origin (Helicobacter pylori) and stress ulcers in cases of thermal injury

Based on these findings the study authors conclude:

"The probiotic products Laktera with Lactobacillus bulgaricus have healing effect in cases of disorders of the gastrointestinal tract of various etiology and severity. Regular intake of the probiotic Laktera helps the maintenance of the activity and vitality of the human body, improves life quality and maintains the gastrointestinal health."

Researchers: Associate professor Dr. Konstantin Ramshev chief of the Department of Intensive therapy and Reanimation, Military Medical Academy, Sofia, Dr. George Alexandrov, Military Medical Academy, Sofia, Department of Surgery and Associate professor Dr. Marieta Georgieva, Ph. d., Medical University, Varna, Department of Pharmacology.

Reference:

  1. Military Medical Academy, Medical University – Sofia, National Center of Hygiene, Medical Ecology and Nutrition, Medical University of Würzburg - Germany, Elbi-Bulgaricum, JSC; “Complex medical-biological studies of the effects of Biostim LBS on preventive and curative processes in people put to extreme overload in case of sea rescue operations, in Air Force pilots and navigators and patients with acute and chronic damages from drugs”; (136 patients)
  2. Military Medical Institute Academy – Sofia, Department of Toxicology and Allergology at the Higher Military Medical Institute – Sofia, Clinic of Toxicology and Allergology at the Naval Hospital - Varna; Exposé of the report for a clinical testing of a low-lactose lactic acid product Biostim LBS in the treatment of patients with various intoxications; (320 patients)
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  4. Military Medical Academy, United Aeromedical Research Institute, Department of Toxicology and Allergology; Exposé of a report and scientific articles for a clinical study of the effect of the bio product Biostim LBS-1 on pilots and navigators from Air Force of the Republic of Bulgaria; (92 patients)
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  15. Department of Microbiology, Medical University of Sofia; Evaluation of inhibitory activity of Lactobacillus delbrueckii subsp. Bulgaricus on Helicobacter pylori growth in vitro; Lyudmila Boyanova, Maria Stephanova-Kondratenko and others; International symposium on Original Bulgarian Yogurt, 2005, Sofia (4 patients);
  16. Biostim LBS: a special-diet and therapeutic low-lactose powdered food product, Military Medical Academy, Sofia, ELBY Engineering-G State Company, Sofia, B. Gyosheva, N. Alexandrov and others, published in Biotechnology & BioE 6/1992/4 Appendix (100 patients);
  17. Scientific practical conference on disasters and working injuries protection, organized by Council of Ministers; 1992 Sofia, Bulgaria; Biostim LBS in case of acute poisoning as a therapeutic food, B. Gyosheva, N. Alexandrov (376 patients);
  18. University of Food Industry – Plovdiv; Dissertation of Petranka Dimitrova Ivanova, Technologies for obtaining of lactic acid foods for babies (0-1 years old) and children with intolerance to milk sugar; Plovdiv, 1991(25 children);
  19. Report in front of Comecon – Council of Mutual Economic Assistance on a symposium Problems of dietary feeding of children, 1989, Sofia; Characteristic of food products for children with enzyme-deficiency; B. Gyosheva and others (20 children);
  20. Military Medical Academy, Sofia, 1991, Dissertation of Nikola Georgiev Alexandrov, Clinical researches on injured people from chemical catastrophes in Bulgaria in the period 1981 – 1990, for conferment of an educational and scientific degree „Doctor of science“ (197 patients);
  21. Minisrty of National defense, meeting of the member states of the Warsaw Pact, 1988, Varna, Report of N. Alexandrov (top secret);
  22. Report on a International congress of nutrition, Varna, 05.2004, Application of probiotics with Lactobacillus bulgaricus in the early postoperative period, G. Alexandrov, D Damianov (64 patients);

Effect of probiotic product Laktera Vision+ with Lactobacillus bulgaricus and extract from bilberry on dry eye syndrome and tired eye syndrome

Effect of probiotic product Laktera Vision+ with Lactobacillus bulgaricus and extract from bilberry on dry eye syndrome and tired eye syndrome

Technology development and penetration of computers in life lead to refractive anomalies of the eyes in more young people and they start to wear glasses or lenses for vision correction at a young age, before the occurrence of age-related changes (presbyopia). Excessive congestion of eyes at a young age led to the occurrence of new syndromes in ophthalmology – dry eye syndrome and tired eye syndrome.

There are lots of myths as well as lots of studies for the positive effect of bilberry on the eye health.

Bilberry (Vaccinum myrthilus) is a perennial shrub with a height up to 35 cm, which grows in forests and mountain regions of Northern Europe, West Asia and North America. Its fruits are small with a black-violet colour and a pleasant sweet sour taste. (1)

The ancient Roman army physician Pedanius Dioscorides (40 AD), who is considered to be the founder of botany, in his book „De materia medica“ recommends bilberry in treatment of diarrhea and scurvy. (2)

100 g bilberry fruit contain 86,5 g water, 1,1 g proteins, 8,6 g carbohydrates, 2,2 g fibres, 1,2 g free organic acids, 51 mg potassium, 6 mg sodium, 16 mg calcium, 6 mg magnesium, 13 mg phosphorus, 7 mg iron, 0,9 mg copper, 0,01 mg Vit. B1, 0,02 mg Vit. B6, 0,3 mg Vit. PP, 10 mg Vit. C. They are very rich in anthocyanides from the group of flavonoids. (1) Anthocyanides are very important for the eye health. Eye lens and retina are exposed to prolonged intense light radiation and singlet oxygen more than any other tissue in the human body. Thus free radicals are formed and they combine with the lipids in the photoreceptor cell membrane (rods and cones), responsible for vision. To keep the eye tissue healthy it must have an effective antioxidant protection. Anthocyanides in the bilberry are powerful antioxidants and help in the fight against cellular degeneration of the retinal cells caused by free radicals. Anthocyanides help to maintain the normal blood flow in the fine blood vessels (capillaries) that feed the eye tissues. (3)

Bilberry is often associated with the improvement of vision at night. The story that during the World War II pilots have taken bilberry jam to sharpen their vision in night missions is very popular. An American clinical study in 2000 on pilots from the U.S. Navy did’t find such effect. (4)

Experimental research of rats show that the use of bilberry extract can inhibit the development of macular degeneration and senile cataracts.(5)

Object of observation were 41 people aged between 21 and 44 years. 24 of them were men and 21 – women. All of them wear glasses for vision correction, as 17 (41.46%) are shortsighted with dioptre from -0.5 to -3d, and 24 (58,54%) are far-sighted with dioptre + 0.5 to + 2d. 28 (68,29%) of them are with astigmatism. 24 (58,13%) of them wear glasses and lenses for vision correction. All of them work in offices (government officials, IT specialists, accountants, etc.) from 4 to 9 hours a day in front of computers an average 5 hours a day. All wear anti-reflecting glasses while working on a computer.

Most frequent syndromes that occur in these people are the dry eye syndrome and the tired eye syndrome. The dry eye syndrome manifests with reduced production of tears, irritation and itching of eyes. The syndrome of tired eye which is a kind of dryness of eyes occurs after continuous work on a computer with red eyes, marking and expansion of capillaries, blurred vision.

The study was carried out within 8 weeks (56 days). 35 of people have taken 2 capsules Laktera Vision + every day before meal. 6 of the observed people have taken Laktera Vision + irregularly – on average 2.5 times a week. Each capsule (320 mg) contains over 20 billion live cells of Lactobacillus delbrueckii spp. bulgaricus, Streptococcus thermophilus, Lactobacillus helveticus, Lactobacillus lactis (isolated from spring water in Bulgaria), 21 amino acids, milk vitamins, minerals and 10 mg dry extract of bilberry (Vaccinium myrtillus), standardized to 25% anthocyanides.

The results from the study are presented in Table 1.

Days % patients with dry eye syndrome % patients with tired eye syndrome
0 70,73 85,36
7 17,07 21,95
14 14,63 19,51
28 12,20 17,07
42 14,63 17,07
56 12,20 14,63

The table shows that the syndromes of dry and tired eye can be put under control very quickly, in the first week after administration of Laktera Vision +. During the eighth week of the study the dry eye syndrome was observed in 12.2 % (5) of the cases and the tired eye syndrome – in 14.63% (6) of the cases. The medical history for those who have not taken the probiotic product with bilberry extract regularly shows that when they have not taken the product for 3-4 days the syndromes occur again. After administration of the product manifestations of the syndromes are put under control for a period of 12 – 24 hours.

Usually the free market offers products which contain in 1 capsule from 5 to 150 mg bilberry extract. In our observation the effect is achieved with 20 mg bilberry extract daily. The smaller necessary daily dose of dry bilberry extract is probably due to a specific amino acid or group of amino acids in the probiotic product, that acts as a transport carrier of the molecules of the extract and potentiate the effect of the anthocyanides in bilberry. Identification of the amino acid or the group of amino acids, responsible for that process, is a subject of further study.

The probiotic product with bilberry extract controls successfully the syndromes of dry and tired eye, occurring in young people wearing glasses for vision correction, who work on a computer more than 4 hours a day.

  1. Serbin AG, Medical Botany, 2003, – S 150. – 364 p. – ISBN 966-615-125-1
  2. Antonova A, Botany // Encyclopedic dictionary Brockhaus and Efron: in 86 volumes (82 items and 4 suppl.) — 1890—1907.
  3. Gross PM, Scientists zero in on health benefits of berry pigments, Natural Products Information Center, July 2007
  4. Muth ER, Laurent JM, Jasper P (April 2000). Alternative Medicine Review 5(2): 164-73. PMID 10767671. “The effect of bilberry nutritional supplementation on night visual acuity and contrast sensitivity”
  5. Furtsova A, Gesarevich DB, Gonchar AM, Trofimova NA, Kolosova NG, 2005, Progress in gerontology, 16:76-9. PMID 16075680


Experimental and clinical researches of Daflorn’s products

1. Military Medical Academy – Sofia, Scientific Institute for Emergency “N. Pirogov” - Sofia; Exposé of a final report entitled „Methods of nutritional therapy – mixed parenteral and enteral nutrition“ adopted by protocol ? 23/1012.1986 by the Scientific Council of the Higher Military Medical Institute - Sofia, Bulgaria

2. Central Experimental Laboratory for production, implantation and design; Contract with Institute for Emergency „Pirogov“ – Sofia for delivery of Biostim – bioproduct for enteral nutrition

3. Central Experimental Laboratory for production, implantation and design; Patent Library AIS „SIREN“, reg. ? 82307019, Report on: “Development of Food for enteral feeding based on freeze-dried lactic acid lactose-free products“, stage II, 3 “Study of amino acid and salt composition of food, a proposal to use the results in practice”

4. Military Medical Academy, Institute for Emergency „N. Pirogov“, Central Experimental laboratory for production, implantation and design; Patent Library AIS „SIREN“, reg. ? 82307019, Final Report on: „Diagnosis and treatment of multiple organ deficiency“, Program task: Development and clinical testing of food and means for enteral feeding on the basis of lyophilized lactic acid lactose-free products

5. Ministry of Health, Institute for Emergency „N. I. Pirogov“; Protocol for the clinical testing of a lyophilized lactic acid product „Biostim“ produced by „Central Experimental Laboratory for production – implantation – design“ at SO „Dairy Industry“ Sofia

6. Military Medical Academy, Institute for Emergency „N. Pirogov“, Central Experimental laboratory for production, implantation and design; Patent Library AIS „SIREN“, reg. ? 82307019, Final Report on: „Diagnosis and treatment of multiple organ deficiency“, Program task: Development and clinical testing of food and means for enteral feeding on the basis of lyophilized lactic acid lactose-free products

7. Military Medical Academy, Medical University - Sofia, National Center of Hygiene, Ecology and Nutrition, Medical University of Würzburg - Germany, Elbi Bulgaricum JSC; Research topic: Complex medico-biological studies of the effects of Biostim LBS on prophylactic and healing processes in people put to extreme strain in sea rescue operations, pilots and navigators from the Air Force and in people with acute and chronic drug damages

8. National Center of Hygiene, Ecology and Nutrition, Medical University of Würzburg - Germany, Military Medical Academy, Medical University – Sofia, SF Elbi; „Complex medical-biological studies in a prolonged vestibular load in case of a shipwreck“; Aim – study of the effect of the prolonged vestibular load and the other experimental sea factors on the body and approbation of the lactic acid product of SF Elbi – Biostim LBB as food with which rescue vehicles in water and air transport can be equipped

9. Command of the Air Force and the Medical Department at the Headquarters of the Air Forces, Air Hygiene Laboratory to OAMNII – MMA; Finding for a study of two lyophilized lactic acid products „Bulgaricum with pectin“ and „Biostim with pectin“, developed on the basis of specific strains of Lactobacillus bulgaricus - manufacturer SF „ELBI ENGINEERING“ - SOFIA; Aim of the task: depending on the results of the study to include these two products in the ration of the flight crews of the Air Force

10. Written statement for the clinical testing of the effect of „Biostim LBB“ on the recovery process in cases of different intoxications in the Clinic of Toxicology and Allergology – MMA Sofia and Clinic of Toxicology – Naval Hospital – Varna

11. Military Medical Academy – Sofia, Department of Toxicology and Allergology at the Higher Military Medical Institute – Sofia, Clinic of Toxicology and Allergology at the Naval Hospital - Varna; Exposé of the protocol for a clinical testing of a low-lactose lactic acid product Biostim LBB in the treatment of patients with various intoxications

12. Contract between Meiji Milk Products Tokyo, Elbi Bulgaricum JSC and Prof. N. Alexandrov for a study on: Complex medical biological studies of the effects of Biostim LBB on lipid metabolism and immune system in people put to extreme mental and physical stress

13. Report of Gen. Major Ass., N. Alexandrov, Ph.D., Col. Dr. R. Zlatev and Col. Dr. V. Minkov to the Command of the Air Force concerning further development of the scientific topic studying the effect of the lactic acid product „Biostim LBB“ on the lipid metabolism and the immune system in pilots and navigators

14. Military Medical Academy, United aero-medical Research Institute, Department of Toxicology and Allergology; Exposé of protocol and scientific articles for a clinical study of the effect of the bioproduct Biostim LBS-1 on pilots and navigators from Air Force of the Republic of Bulgaria

15. Department of Preclinical and Clinical Pharmacology and Biochemistry, Medical University - Varna; Department of Toxicology and Allergology, MMA - Sofia; University Hospital „St. Anna – Varna“ JSC, Hospital Pharmacy; Clinical Laboratory, Hospital „St. Marina“ - Varna; Department of Nephrology, Haemodialysis and Toxicology, Hospital „St. Marina“ - Varna – Study of the effects of „Biostim LBS“ and silymarin on an experimental model of carbon tetrachloride induced hepato-toxicity in rats

16. Department of Preclinical and Clinical Pharmacology and Biochemistry, Medical University - Varna; Department of Toxicology and Allergology, MMA - Sofia; University Hospital „St. Anna – Varna“ JSC, Hospital Pharmacy; Clinical Laboratory, Hospital „St. Marina“ - Varna; Study of the effect of the probiotic Biostim LBS on the lipid metabolism

17. Department of Preclinical and Clinical Pharmacology and Biochemistry, Medical University - Varna; Department of General and Clinical Pathology, Medical University - Varna; Department of Toxicology and Allergology, MMA - Sofia; University Hospital „St. Anna – Varna“ JSC, Hospital Pharmacy; Clinical Laboratory, Hospital „St. Marina“ - Varna; Study of the effect of the probiotic Biostim LBS in experimentally induced hyperlipidemia

18. Medical Faculty – Sofia, Department of Pharmacology; Written statement for a comparative pharmacological and biochemical study of the effects of Laktera, Laktera/Panax and Laktera /Hypericum on ulcer formation and ethanol-induced oxidative stress in rats

19. Department of Preclinical and Clinical Pharmacology and Biochemistry, Medical University - Varna; Department of General and Clinical Pathology, Medical University - Varna; Protective effect of the probiotic Biostim LBS in a model of indomethacin-induced ulcer-genesis

20. Department of Preclinical and Clinical Pharmacology and Biochemistry, Medical University - Varna; Department of Pathophysiology, Medical University – Varna; Department of Anatomy, Histology, Embryology, Medical University - Varna; Effects of the probiotic Biomilk on indomethacin-induced oxidative damage of some tissues

21. Department of Preclinical and Clinical Pharmacology and Biochemistry, Medical University -Varna; Department of General and Clinical Pathology, Medical University – Varna; Department of Toxicology and Allergology, MMA - Sofia; Effect of the probiotic Biostim LBS on histopathological changes in liver in experimental hypercholesterolemia

22. Department of Preclinical and Clinical Pharmacology and Biochemistry, Medical University - Varna; Department of General and Clinical Pathology, Medical University – Varna; Department of Toxicology and Allergology, MMA - Sofia; Hepato-protective effect of the probiotic Biostim LBS on experimental model of carbon tetrachloride-induced hepatotoxicity in rats

23. Department of Preclinical and Clinical Pharmacology and Biochemistry, Medical University - Varna; Department of General and Clinical Pathology, Medical University – Varna; Department of Toxicology and Allergology, MMA - Sofia; Department of Radiology, Medical University – Varna; Ray-protective effect of the probiotic Biomilk containing Lactobacillus bulgaricus

24. Abstract of Dissertation of Daniela Petrova Peneva for conferment of an educational and scientific degree „Doctor“

25. Abstract of Dissertation of Marieta Petrova Georgieva for conferment of an educational and scientific degree „Doctor“

26. Statement of Colonel Dr. Vencislav Minkov Minkov, Head of „Medical Department“ at division 22800 to the Chief of the General Staff of the Air Force for a study of two products – Bulgaricum with pectin and Biomilk with pectin used by pilots from the Air Force

27. MMA, An extract from the minutes from Vice Head of MMA for educative and scientific affairs, Ass. Prof. Dr. A. Andreev on ongoing clinical tests of low-lactose dry milk products with trademarks „Biomilk“ and „Laktera“

28. An opinion of the Head of the Department of Toxicology and Allergology at the Military Medical Academy - Sofia, Prof. Dr. Nikola Alexandrov Ph. D., about replacing yogurt with wholefood supplement with probiotic effect Biomilk and Biostim in conditions harmful for health.

29. Certificate of MMA to give “Biomilk” and “Laktera”, produced by Daflorn Ltd., as wholefood supplements and functional food to the contingent of the Bulgarian Army. And their use in Military Medical Academy for more than 10 years as a food therapy of seriously ill and operated patients.

30. MMA, Head of the Clinic of Toxicology and Allergology, Associate Dr. N. Alexandrov; Recommendation for the use of probiotics in cases of serious toxicological and allergic diseases (Epidermolis necroticans bulosa Syndrome), as well as in other cases of toxicological and allergic diseases with beneficial effect.

31. MMA, Head of the Clinic of Toxicology and Allergology, Associate Dr. N. Alexandrov; Protocol for the use of the products from the series „Biomilk“ and „Biostim“, containing Lactobacillus Bulgaricus

32. MMA, Deputy for the health affairs of MMA, Ass. Prof. Ramshev; Recommendation for the use of different probiotics with trademark Biomilk and Laktera with a beneficial effect in patients requiring stimulation of the immune system of the body because of different viral or bacterial diseases.

33. Statement of the Ministry of Labour and Social Affairs to the Ministry of Health and the General Headquarters of the branch syndicates – Proposal to include the wholefood supplement Laktera in the nomenclature of the antidotes and protective food

34. Recommendation from Alexander Chakmakov, Ex. Director of „Titan Zlatna Panega Cement“, JSC for the benefit of the probiotics Biomilk and Laktera as a protective food

35. MMA, Certificate for clinical tests and Certificates of Analysis of the probiotic wholefood supplements Biomilk, Laktera and Magic Power.

36. Certificate of MMA to give “Biomilk” and “Laktera”, produced by Daflorn Ltd., as wholefood supplements and functional food to the contingent of the Bulgarian Army located in Iraq. As well as their use in the Military Medical Academy for more than 10 years as a food therapy of seriously ill and operated patients.

37. Full Doping control of the product series Biomilk in German Sports University of Cologne – Doping Laboratory

38. Ukrainian Research Institute of Medical Transport, State Department of maritime and river transport of Ukraine, Medical University - Varna – Effects of Biomilk Hepanorm in simulated shipwreck

39. Journal „Medicine and Health“ - „Ray-protective effect of a lactic acid product containing a Lactobacillus bulgaricus strain“

40. Annual book of IMAB for Bulgaria, Israel, Germany, Norway, France, Italy and USA – Study of the probiotic Biostim for teratogenic effect

41. International Conference in Lyon – Lactic acid fermentation and biotechnological method of obtaining a low-lactose lactic acid product for special diets and therapeutic food

42. Journal „Biotechnology“ - „Effect of the low-lactose lactic acid products Bulgaricum and Biostim on the immune system“

43. National Cancer Center, National Centre of Haematology and Transfusiology; Study of the ray-protective effect of Gerisan and Biostim LBS on the basic leukocyte populations in cases of radiotherapy of cancer.

44. Balkan Military Medical Committee; The 4th Congress of B.M.M.C.; June 6-10, 1999, Bucharest, Romania; EFFECT OF THE BIOSTIM LBB YOGHOURT-TYPE PRODUCT ON LIPID METABOLISM IN AIR-FORCE AVIATORS; Author(s): Alexandrov, N., Petrov, A., Zlatev, R., Zhekova, M., Mikhaylova, Z., Kotseva, M. and Alexiev, L. Institution: Military Medical Academy, Sofia Institution For Aviation Medicine;

45. FEBS; 20th MEETING OF THE FEDERATION OF EUROPEAN BIOCHEMICAL SOCIETIES, organized by the Hungarian Biochemical Society; Budapest, Hungary, August 19-24, 1990; Freeze dried dietetic lactic acid product “Biostim LBS”; Gyosheva B., Alexandrov N., Ivanova P., Manafova N., Georgiev G.; Central Exper. Lab. Dairy Ind. 1618 Sofia; Emergency Inst. “N. Pirogov” 1612 Sofia, Bulgaria.

46. An International Journal for the Rapid Publication of Short Reports on Biochemical Mechanisms of Mammalian Toxicity; Clinical Research on the Victims of Chemical Accidents in Bulgaria; N. Alexandrov, D. Jordanova and M. Georgieva, Military Medical Academy, Sofia, Bulgaria; VI INTERNATIONAL CONGRESS OF TOXICOLOGY, Rome, 28 June – 3 July 1992

47. Therapeutic Drug Monitoring; A Journal Devoted to Therapeutic Drug Monitoring and Clinical Drug Toxicology, volume 15, number 2, 1993, Philadelphia, USA; Influence of Nootropic Agents and Hyperbaric Oxygenation in the Treatment of Acute Poisonings with Halogenated Hydrocarbons; N. Alexandrov, M. Georgieva, Military Medical Academy, Sofia, Bulgaria.

48. Addictive drugs and addictive states, The state of the art; Addiction Research Foundation of Italy (ARFI), Padova in collaboration with University of Florence – Department of Preclinical and Clinical Pharmacology “Mario Aiazzi Mancini”; International congress of Toxicology in Florence, 07.1992; Clinical research on narcotic and non-narcotic drug addiction. Comparative analysis; N. Alexandrov, M. Georgieva, D. Jordanova.

49. Biotechnology Journal; “Biostim LBS – a special diet and therapeutic low-lactose powdered product”, issue 4, series V, 92 – english version. Biostim LBS – a special diet and therapeutic low-lactose powdered product; B. Gyosheva, N. Alexandrov, N. Manafova, P. Ivanova, G. Georgiev; ELBY Engineering –G, Sofia and Military Medical Academy, Sofia, Bulgaria.

50. Biotechnology Journal, 2/1994; Effect of dairy products, containing Lactobacillus strain LBL4 on some biochemical parameters in humans; M. Zhekova¹, K. Avramova², B. Gyosheva³; Aviomedical Research Institute, Higher Medical Academy, Sofia; National Center of Hygiene, Medical Ecology and Nutrition, Sofia; ELBY Engineering-G, Sofia.

51. Biotechnology Journal, 4/1992; Biostim LBS: a special diet and therapeutic low-lactose powdered food product; B. Gyosheva, N. Alexandrov, N. Manafova, P. Ivanova, G. Georgiev; ELBY Engineering – G State Company, Sofia; Military Medical Academy, Sofia.

52. Cancer Letters Journal 114 (1997) 93-95; Preventive role of lyophilic dairy products bulgaricum and biostim; T. Ivanova, M. Jekova; Transport Medical Institute, Sofia, Bulgaria; Medical Military Aviation Institute, Sofia, Bulgaria.

53. Contract between Joint MLB Promotion Planning Committee between MEIJI MILK, Tokyo and LB Bulgaricum PLC, Sofia and Prof. Nikola Georgiev Alexandrov D.Sc. – Head of Toxicologic and Allergologic Department at Military Medical Academy; Clinical-Immune Researches over some effects of the yoghurt product “Biostim LBB” on pilots from the Army-Air Forces; N. Alexandrov¹, K. Plotchev², K. Penkova², R. Vladimirova², K. Mladenov², P. Zlatev³, Dzh. Nedyalkova; ¹ Military Medical Academy, Sofia – CLINIC OF TOXICOLOGY; ² Military Medical Academy, Sofia – CENTRAL LABORATORY ON CLINICAL IMMUNOLOGY; ³ OAMNII, Military Medical Academy, Sofia.

54. Fundamental & Clinical Pharmacology; The 3rd Meeting of the Federation of the European Pharmacological Societies (EPHAR); 6-9 July 2001, Lyon, France; Effects of the chronic dietary supplementation of the probiotic Biostim LBS on carbon tetrachloride-induced hepatotoxicity in rats; K. Marazova¹, N. Alexandrov², ¹Department Pharmacology Toxicology, Medical Faculty, Sofia, Bulgaria, ²Department Toxicology Allergology, Military Medical Academy, Sofia, Bulgaria.

55. Journal „Medicine and Pharmacy“, issue 2 / 2002, „Clinical features and treatment of acute intoxications with heroin”

56. Journal „Medicine and Pharmacy“, issue 4 / 2003, „Treatment of organophosphorus pesticides poisoning“

57. „Medicine and Pharmacy“, issue 2 / 2003, „Study of the probiotic Biostim LBS in experimentally induced hyperlipidemia“

58. VIIth National Congress of Nutrition with International participation; Nutrition science for human health protection; Varna – St. Konstantin and Elena, Bulgaria, May 15-17, 2003; Probiotics application before and in the early post-operative period; Alexandov G., D. Damianov, Clinic of Surgery, UNBAL “Queen Jovana” - Sofia

59. NATO Symposium; May 2005, Munich, Germany; Complex treatment of organophosphorus pesticide poisoning as a model of chemical warfare agents; Alexandrov N., Ks. Stoyancheva, Military Medical Academy, Toxicology Department.

60. International Symposium on original Bulgarian yogurt under the patronage of President of the Republic of Bulgaria, Sofia, 2005 ; The application of probiotics for restoration of vaginal environment; Stefan Kovachev¹, Aneta Nacheva¹, Daniela Petrova², Milena Kovacheva²; ¹MC “Gynart” MMA – Sofia; ²MC “Evrozdrave” MMA – Sofia

61. International Symposium on original Bulgarian yogurt under the patronage of President of the Republic of Bulgaria, Sofia, 2005; Farmacological study of the probiotic Biomilk, containing Lactobacillus bulgaricus, for antioxidant activity, Georgieva M.¹, Bekyarova G.², Alexandrov N.³, Manolov N.?; ¹Department of preclinical and clinical pharmacology and biochemistry, Medical University “Prof. Dr. Paraskev Stoyanov” – Varna, ²Department of Pathophysiology, Medical University – Varna, ³Department of toxicology and allergy, MMA – Sofia, ?Clinic of Vascular Surgery, University Hospital “St. Anna” – Varna

62. International Symposium on original Bulgarian yogurt under the patronage of President of the Republic of Bulgaria, Sofia, 2005; Good bacteria of Bulgarian youghurt – a base of symbiotic and functional food, Alexandrov N.¹, Gyosheva B.², Ivanova P.³; ¹Military Medical Academy, ²Bulgarian Association of Dairy Processors, ³LB Bulgaricum

63. International Symposium on original Bulgarian yogurt under the patronage of President of the Republic of Bulgaria, Sofia, 2005; Probiotics – base of functional nutrition and nutritive treatment of XXI century; Nikola Alexandrov¹, Phd, Daniela Petrova², Milena Kovacheva¹; ¹Emergency Clinic of Toxicology and Allergy, MMA – Sofia, Bulgaria, ²MC “Evrozdrave” MMA Sofia, Bulgaria

64. International Symposium on original Bulgarian yogurt under the patronage of President of the Republic of Bulgaria, Sofia, 2005; Probiotics – lifestyle, M. Georgieva¹, E. Softova², P. Borisova, N. Alexandrov³, N. Manolov?; ¹Department of preclinical and clinical pharmacology and biochemistry, Medical University “Prof. Dr. Paraskev Stoyanov” – Varna, ²Department of General and Clinical Pathology, Medical University “Prof. Dr. Paraskev Stoyanov” – Varna, ³Department of Toxicology and Allergy, MMA – Sofia, ?Clinic of Vascular Surgery, University Hospital “St. Anna” – Varna

65. International Symposium on original Bulgarian yogurt under the patronage of President of the Republic of Bulgaria, Sofia, 2005; Synbiotics – their place in modern treatment of abusers, Daniela Petrova¹; Milena Kovacheva²; ¹MC “Evrozdrave” – MMA, Sofia, ²Clinic of Emergency Toxicology and Allergy, MMA – Sofia

66. Journal „Military Medicine“, issue 4 / 2007, „Study of the antioxidant activity of the probiotic Biomilk under experimental conditions“

67. Journal „Military Medicine“, issue 2 / 2007, „Ray-protective effect of the probiotic Biomilk“

68. Journal „Medicine and Pharmacy“, issue 9-10, „Alcoholic disease, clinic and treatment“

69. Journal „Medicine and Pharmacy“, issue July/ 2008, „Probiotics and vaginal flora”

70. Journal „Medicine and Pharmacy“, issue 1 / 2009, „Prevention and treatment of atherosclerosis with probiotic lactic acid food and bacteria“

71. EXPERIMENTAL CLINICAL STUDY OF THE EFFECT OF THE PROBIOTIC BIOMILK IN CASES OF LIVER INJURIES; VI National Congress of Pharmacology 2009; Georgieva M.¹, Popov G.², Plochev K.², Alexandrov N.²; ¹ Department of Preclinical and Clinical Pharmacology and Toxicology, Medical University of Varna; ² Military Medical Academy

72. Effect of a probiotic product with Lactobacillus bulgaricus DWT1 in avoiding adverse effects during adjuvant treatment of patients with tumors of the stomach, colon and rectum. Medical University, Zagreb, Republic of Croatia, Pimezon Polyclinic; Prof. Dr. Mirko Shamija; December 2012

73. Magazine. Forensic Medicine 9/2013, A meta-analysis of the effects of a probiotic product with Lactobacillus bulgaricus Laktera in the gastrointestinal tract; Ramshev C. 1 D. Alexandrov 1; 1 MMA Sofia

74. MU Varna power. Magazine., 2013, Effect of probiotic product Laktera Vision with Lactobacillus bulgaricus and bilberry extract on syndromes dry and tired eyes Georgieva M.1 Alexandrov D.2; 1 MU Varna, 2 Sofia MMA

75. World Congress of probiotics Athens 2014, newly origin of probiotics in the Balkans; Petrov N.1 G. Alexandrov 1 Peneva M. 2; 1 MMA Sofia 2Plovdiv University of Plovdiv

76. Bulgarian Red Cross 2014 Recommendation on the use of probiotic foods Laktera of victims of disasters and accidents.

77. World Journal of Pharmacy and Pharmaceutical Sciences, volume 4, issue 06, 2015q 130-136, Antiproliferative Effect of Bulgarian Spring Water Probiotics (Laktera Nature Probiotic®) Against Human Colon Carcinoma Cell Line, Kaloyan Georgiev, Marieta Georgieva, Ivan Iliev, Maria Peneva, Georgi Alexandrov; Department of Preclinical and Clinical Sciences, Sector Pharmacology and Toxicology, Faculty of Pharmacy, Medical University Varna; Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences, Sofia; Medical Centre Evrozdrave Sofia; Military Medical Academy Sofia.

78. 6th Southeast European Conference of Chemotherapy, Infectious and Cancer Diseases, November 2015, Thessaloniki, Greece; Anti-tumor Effect of probiotics – experimental and clinical research, Georgi Alexandrov, Kaloyan Georgiev, Marieta Georgieva, Ivan Iliev, Maria Peneva; Military Medical Academy Sofia; Department of Preclinical and Clinical Sciences, Sector Pharmacology and Toxicology, Faculty of Pharmacy, Medical University Varna; Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences, Sofia; Medical Centre Evrozdrave Sofia.

79. 3rd Biennial Conference of Probiotic Association of India (PAi), International Symposium on Stress, Microbiome & Probiotics, March 11-13, 2016, NISER, Bhubaneswar, India; Application of Lactobacillus bulgaricus in Clinical Practice; Georgi Alexandrov, Maria Peneva; Military Medical Academy Sofia; Medical Centre Evrozdrave Sofia.

80. Excerpt from Abstract of dissertation, Sofia 2016, p. 27-29; Epidemiological study of patients with food allergies; Maria Peneva, Medical Centre Evrozdrave Sofia.

81. Scripta Scientifica Pharmaceutica, ISSN 2367-6000, volume 4, 2017, supplement 2, page 51, Medical University Prof. Dr. Paraskev Stoyanov, Varna, Bulgaria, "Hepatoprotective effect of probiotic food, containing lactobacillus bulgaricus DWT1 in acute ethanol intoxication in experimental animals", Gabriela Kehayova1 , Marieta Georgieva, Kaloyan Georgiev, G. Aleksandrov, M. Peneva.

82. World Journal of Pharmaceutical Research, volume 7, Issue 12, 2018, Research Article, page 35-42, "Hepatoprotective effect of probiotic, containing Lactobacillus bulgaricus DWT1, in acute paracetamol-induced liver damage in rats", Gabriela Kehayova, Marieta Georgieva and Kaloyan Georgiev, Medical University, Varna

83. Monography, Varna 2018, publishing house Steno, pages 167, "Probiotics - Medicine and Food", Marieta Georgieva

84. Journal of Functional Foods, Volume 56, May 2019, Pages 232-245, “A probiotic formulation containing Lactobacillus bulgaricus DWT1 inhibits tumor growth by activating pro-inflammatory responses in macrophages”, Dipanjan Guhaa, Arka Banerjeea, Raktim Mukherjeea, Biswaranjan Pradhana, Maria Peneva, Georgi Aleksandrov, Sujit Suklabaidya, Shantibhushan Senapati, PalokAich,


Nutritional therapy for oncological diseases

Effect of a probiotic product containing a strain of Lactobacillus bulgaricus DWT1 on preventing adverse effects during adjuvant therapy for patients with tumours of the stomach, colon and rectum

Summary

Previous experience in oncology. The studies of some strains of Lactobacillus bulgaricus have shown that they have antimutagenic effect. Some experiments on animals show a certain beneficial effect in cases of colorectal cancer in rodents. A population study noted lower levels of occurrence of colorectal cancer among a considerable group of fermented dairy products consumers.

Tumours in the digestive system are of exclusive importance because of their high frequency rate and because of the great importance of the digestive system for the whole body. Nutritional therapy that improves digestion is very important for the treatment of the underlying disease as well as for the symptomatic treatment. On the other hand, adjuvant therapy, i.e. the specific oncological treatment /chemo- or radiotherapy/ often leads to a number of healthy tissue damages and to serious complications and side effects. These complications inhibit the beneficial effects of treatment and even make it impossible. Cytostatics cause nausea and vomiting during and after application of chemotherapy and overtaking vomiting due to inadequate antiemetic prevention in previous chemotherapy. Mucositis is an extremely serious side effect after both chemotherapy and irradiation in the area of head and neck. It concerns inflammatory changes of the mucous membrane of the digestive system. It often occurs in parallel with myelosuppression and leukopenia. The most dangerous consequence of leukopenia /decrease in the number of leukocytes/ is febrile neutropenia that requires hospitalization, blood transfusion and often leads to discontinuation of cytostatics and the quality of life is getting worse. Anorexia and cachexia /weight loss/ are frequent in patients with tumors of the digestive system and adversely affect the successful treatment, tolerance of treatment and indirectly affect survival. Radiotherapy causes adverse effects depending on the total dose, fraction size, field of irradiation, interval between fractions, type of radiotherapy and specificity of the irradiated tissues. The toxic effects of radiotherapy are dyspepsia, gastritis, ulceration /ulcers/ and radiation enteritis.

Results. The study included 80 patients with tumours of the stomach, colon and rectum. All of them have undergone a patho-histological examination and the disease stage was precisely defined. In this randomized study patients were divided into two groups of 40 patients. The first group of 40 patients was undergoing chemotherapy or chemoradiotherapy. The other group of 40 patients was taking probiotic food, containing strain of Lactobacillus bulgaricus DWT1 – 15 mg once a day before breakfast for three months, in addition to the chemotherapy or chemoradiotherapy.

The obtained results show that probiotics, containing a strain of Lactobacillus bulgaricus DWT1, have a potential beneficial effect as taken in addition to the treatment of cancer patients with adjuvant chemotherapy or chemoradiotherapy. Administration of that probiotic product influences on the reduction of nausea and vomiting and maintains appetite of patients during the adjuvant therapy. There is a clear bond between reduction of the frequency and severity of disorders and the intake of Lactobacillus bulgaricus DWT1.

Nutritional therapy with probiotic product, containing a strain of Lactobacillus bulgaricus DWT1, helps the prevention of the adverse effects during adjuvant therapy.

Researchers: Prof. Dr. Mirko Shamia, PhD and associates, Zagreb, December 2012