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)

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)

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)

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)

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

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.

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.

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).

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.

Our results obtained in 152 patients are presented in figure 5.

It is obvious that regular use of the probiotics Biomilk and Laktera in combination with the main treatment of alcohol dependence leads to control of consumative and bone articular syndrome and restoration of trophic of skin and hair for a period of about 1 year.

Anti-tumor and anti-mutagenic effect of the lactic acid bacteria

Factors that lead to cancer are three – carcinogens, mutagens and microorganisms. The process of cancer formation occurs in two stages – appearance of mutant cells as a result of damage of the cellular DNA and transformation of mutant cells into cancer cells. This process is shown in figure 6.

Figure 6 Process of cancer formation due to chemical agents

Factors leading to cancer exist in our life and we are constantly exposed to their harmful influence. Cancer cells are constantly formed in the human body, but phagocytes destroy them. When the immune system weakens, especially in combination with stress, the organism cannot respond adequately to the increased number of cancer cells and they begin to multiply uncontrollably.

Some studies, conducted in the Agricultural department of Shinshu University in Japan, show that lactic acid bacteria have anti-mutagenic effects. Table 7 shows the anti-mutagenic effects of different lactic acid bacteria and food against the mutagen AF2 .

Table 7

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”

Preliminary announcement for nutritional therapy with probiotic food Laktera, containing strains of Lactobacillus bulgaricus DWT1 and Streptococcus thermophilus DWT4 in patients with COVID-19

We established in vitro and in vivo efficacy of probiotic cocktail containing Lactobacillus bulgaricus DWT1 and Streptococcus thermophilus DWT4 of water origin. We showed that the probioic cocktail could polarize macrophages from M2 to M1 state. The study was done in collaboration with Prof. Palok Aich of National Institute of Science Education and Research (NISER), India. The results further revealed that the strains in the cocktail could control  tumor growth 

The Lecture of Prof. Palok Aich on the subject you can watch here.

Recently we applied a nutritional therapy with probiotic Laktera, containing the strains Lactobacillus bulgaricus DWT1 and Streptococcus thermophilus DWT4, to patients with COVID-19. The probiotic Laktera is produced by the Bulgarian company Daflorn Ltd and has a certificate by the Bulgarian Food Safety Agency for special purpose foods.

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. Before that the patient has been in COVID ward for 33 days and then discharged with a negative PCR for COVID-19. A few days later, a rash appeared and the patient was admitted on 18.05.2020 again in severe general condition with generalized rash all over the body, including the feet and genitals. This is the first case of COVID-19 with such clinical picture and this type of rash described in Bulgaria. There are scientific data from Italian and English doctors who describe cases of COVID-19 with a similar type of rash. Most likely, this rash is due to damage of the endothelium and arterioles. Photo section 1 / 22.05.2020 are taken on May 22^nd 2020 where the patient is treated with conventional therapy for COVID-19 without probiotic application. The rash affected not only the skin of the whole body, but also the mucosa, and there was an expressed oral necrosis. The patient was not able to eat.

Another 72 patients with positive PCR test for COVID-19 were admitted in the same University hospital in Bulgaria. In them, the clinical picture is manifested by respiratory failure, fever and radiographic evidence of pneumonia. Most of the patients have concomitant diseases – diabetes, obesity, arterial hypertension, metabolic syndrome, Hashimoto’s disease, etc. From the first day of hospitalization, all these patients received nutritional therapy with probiotic Laktera, containing strains Lactobacillus bulgaricus DWT1 and Streptococcus thermophilus DWT4, in a dose of 15 g twice a day in addition to the standard treatment for COVID-19. It is noteworthy that, unlike other patients with COVID-19 pneumonia, those taking the probiotic, clinically recover much more rapidly. On the third-fourth day of hospitalization, they are in good general condition, afebrile, without signs of respiratory failure and severely reduced cough. PCR tests for COVID-19 of these patients were negative and pneumonia was resorbed or in resorption on the 9-10th day of hospitalization. There were no deaths in patients with COVID-19 receiving nutritional therapy with Laktera probiotic. It is obvious from the performed immunological tests that most patients are admitted with high values of T-helper cells, low values of T-cytotoxic lymphocytes and low values of NK-cells. After taking the Laktera probiotic at a dose of 15 g twice a day T-helper cells are reduced. They produce cytokines and their reduction avoids a cytokine storm. T-cytotoxic lymphocytes and NK cells are increased and during the therapy with Laktera they have high levels. The results are presented in Chart 1.

Patients were also tested for immunoglobulins IgA, IgM,IgG and IgE and no abnormalities were observed.

Observation of patients continues after discharge from the hospital till now.

To date, none of the medical staff taking care of COVID patients got infected. The staff is taking Laktera at a dose of 15 g per day.

We have similar observations in over 20 families. One of the families consists of seven members – two small children and three members over 70 years old with concomitant diseases. Two of them over 70 years and a woman of 60 years are taking Laktera probiotic. Four members of the family are infected with COVID-19 and one woman over the age of 70 died. The other three who have been taking Laktera (two of them over the age of 70 and a woman of 60 years) at a dose of 15 g per day for several years, did not get sick from COVID-19, although they have a close contact with the other family members.

Clinical observations continue.

Our clinical research confirms the experimental in vitro and in vivo studies conducted by Prof. Palok Aich and the importance of Lactobacillus bulgaricus DWT1 and Streptococcus thermophilus DWT4 for repolarization of macrophages from M2 to M1 state and enhancing the innate immunity of the human body.

Clinical observations and immunological studies in patients with COVID-19 show that Laktera modulates cellular immunity by reducing T-helpers and by increasing T-cytotoxic lymphocytes and NK cells.

The Laktera probiotic, containing the strains Lactobacillus bulgaricus DWT1 and Streptococcus thermophilus DWT4 of water origin, is an effective nutritional therapy as part of a complex treatment regimen in patients with COVID-19.

Observations of the medical staff working in COVID and sorting wards, as well as of families, indicate that the immunomodulator Laktera can be used as a prevention in the fight against COVID-19.

Date: 16.12.2020, Sofia, Bulgaria

Ret. M. Gen. Prof. Nikola Alexandrov, MD, PhD, D. Sc.

Assoc. Prof. Dr Evgeniq Burzashka MD, PhD

Dr. Daniela Petrova MD Ph.D

Dr. Georgy Aleksandrov MDPh.D

Maria Peneva Ph. D

Chart 1 Cell immunity in COVID-19 patients, treated with Laktera nutritional therapy

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.

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.

Lactobacillus Bulgaricus statement

Magnesium statement

Magnesium statement 2

Oli Ola statement

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. 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,
85. Probiotics in the Prevention and Management of Human Diseases,Academic Press, 2022, Pages 431-448, Chapter 29 – Probiotics: past, present, and future challenges, Marieta Georgieva, Kaloyan Georgiev, Nadezhda Hvarchanova,
86. Educational event III international science conference “Human microbiota” FSBSI “Institute of experimental medicine”, October 7-8, 2021, St. Petersburg, “Newly found origin of probiotics in the Balkans” , Nikolai Petrov, Georgi Aleksandrov, Maria Peneva
87. National Conference on Clinical Toxicology with International Participation on “Toxicology Today“ June 2-4, 2022, Pleven Medical University, “Probiotics – contemporary challenges”, Marieta Georgieva, Georgi Alexandrov, Maria Peneva, Daniela Petrova
88. National Conference on Clinical Toxicology with International Participation on “Toxicology Today“ June 2-4, 2022, Pleven Medical University, “MORBUS LYELL – clinical case”, Katerina Stefanova, Evgenia Burzashka, Olimpiada Atmazhova, Ani Aleksandrova, Nadya Vasileva,
89. National Conference on Clinical Toxicology with International Participation on “Toxicology Today“ June 2-4, 2022, Pleven Medical University, “Toxic-necrotic changes after SARS-COV 19 a clinical case”, Olimpiada Atmazhova, Evgenia Burzashka, Katerina Stefanova, Ani Aleksandrova, Nadya Vasileva, Vladislav Tenev
90. 11^th National Congress of nutrition with international participation “Nutrition science with evaluation of the present and look to the future”, 2022, Bulgarian Society of nutrition and dietetics, “Antioxidant and hepatoprotective activity of probiotic food against paracetamol-induced liver damage in rats”, M. Georgieva, K. Georgiev, N. Hvarchanova, S. Stoeva, M. Radeva-Ilieva, V. Tzankova, M. Kondeva-Burdina, D. Aluani, M. Tzaneva
91. Patent No. 4294 dated 06.07.2022, registered in the Patent Office of the Republic of Bulgaria for a biologically active product used as nutritional therapy in the treatment of patients with COVID-19, including pregnant women.
92. Scientific project No. 21-00038 to study the immune- stimulating and protective effect of probiotic food Laktera in extreme stress situations during the upcoming trip, stay and return of our expedition to Antarctica on Livingston. The project is carried out by the VVMU “N. Y. Vaptsarov” together with the University of Varna and the University of St. Kliment Ohridski” for the period 2022-2023.
93. 16th International Scientific Conference on Probiotics, Prebiotics, Gut Microbiota and Health – IPC2023, 20-22 June 2023, Bratislava, Slovakia, Application of a probiotic formula containing Lactobacillus bulgaricus DWT1 in patients with COVID-19, Maria Peneva, Ioannis Misopoulos, Evgenia Burzashka, Tsvetan Lukanov, Tsetsa Doichinova, Anna Petkova, Marieta Georgieva, Georgi Alexandrov, Pencho Tonchev, Medical Center “Eurohealth-Bulgaria”,  Sofia, Bulgaria, Mika Nutri-Pharma OOD, Petrich, Bulgaria, UMBAL Pleven “Dr. Georgi Stranski”, Pleven, Bulgaria, University Multispecialty Hospital for Active Treatment St. Anna Sofia, Sofia, Bulgaria, Varna Medical University “Prof. Dr. Paraskev Stoyanov”, Varna, Bulgaria
94. V International Scientific Conference ”Microbiota of Humans and Animals”, Institute of Experimental Medicine, October 11-13, 2023, St. Petersburg, Effects of Lactobacillus bulgaricus DWT1 on cell immunity in patients with COVID-19, Georgiy Alexandrov, Maria Peneva, Ioannis Misopoulos, Evgenia Barzashka, Tsvetan Lukanov, Tsetse Doichinova, Anna Petkova, Marieta Georgieva, Pencho Tonchev
95. National Clinical Toxicology Conference “Toxicology in the lens of the present and the future”, November 2-4, 2023, Bulgaria, Probiotics and modulation of cellular immunity in COVID-19, Olympiada Atamadjova, Katerina Stefanova, Ani Alexandrova, Georgi Alexandrov, Maria Peneva, Anna Petkova, Marieta Georgieva, Pencho Tonchev