The team of LAKTERA got a new PATENT for the strain of water origin Lactobacillus bulgaricus DWT1 as a PSYCHOBIOTIC. The products LAKTERA decrease depression, anxiety and improve physiological sleep under stress.

Results of a clinical trial of Lactobacillus Bulgaricus DWT1 in patients with COVID-19 presented at the 16th International Scientific Conference on Probiotics, Prebiotics, Gut Microbiota and Health – IPC2023, June 20-22, 2023 in Bratislava, Slovakia

You can see the entire presentation below the abstract

Abstract

Application of Probiotic Formula, Containing Lactobacillus Bulgaricus DWT1 in Patients with COVID-19

Mariya Peneva^a1, Ioannis Misopoulos^b,Evgenia Barzashka^c1, Tsvetan Lukanov^c2, Tsetsa Doichinova^c3, Anna Petkova^d, Marieta Georgieva^e, Georgy Aleksandrov^a2, Pencho Tonchev^c4

• a – Medical Center “Eurozdrave-Bulgaria”, 7 Rusaliisky prohod str., Sofia, Bulgaria
• b – Mika Nutri-Pharma Ltd., 18 Samuil str., Petrich, Bulgaria
• c – University Hospital Pleven “Dr. Georgy Stranski”, 8a Georgy Kochev blvd., Pleven, Bulgaria
• d – University General Hospital for Active Treatment St. Anna Sofia, 1 Dimitar Mollov str., Sofia, Bulgaria
• e – Medical University of Varna “Prof. Dr. Paraskev Stoyanov”, 55 Marin Drinov str., Varna, Bulgaria

COVID-19 pandemic affected the world and claimed over 6 million victims. That’s why this clinical study shows significant information. Aim of the study is to evaluate the effect of probiotic formula, containing Lactobacillus bulgaricus DWT1, Streptococcus thermophilus DWT4, on cellular immunity in COVID-19 patients.

PCR tests, paraclinical blood tests, biochemical and immunological tests, X-ray, lung CT and medical consultations were performed. Statistics is done by IBM SPSS 23version.

The study is prospective, cohort, randomized, open-label. The subject are 78 inpatients treated for COVID-19 in 2020 randomly divided into two groups: group A – 47 patients taking probiotic, containing Lactobacillus bulgaricus DWT1 at daily dose 15-45g as part of complex therapy for COVID-19; group B – 31 patients receiving only standard therapy. Statistically the two groups didn`t differ in gender, age, incidence, type of concomitant diseases. Treatment in all cases included infusions, vitamins, antipyretics, corticosteroids, anticoagulants, H2-blockers, combination of antibiotics, oxygen. Treatment of group A patients included additional nutritional therapy with probiotic, containing Lactobacillus bulgaricus DWT1. Comparison of cellular immunity activity results in both groups showed statistically significant increase in NK-cells values, increase in cytotoxic T-lymphocytes marker and decrease/ normalization of T-helpers marker in group A patients. T-helpers produce cytokines which at high levels cause cytokine storm and death. Deaths in group B were patients admitted with high T-helpers, which increased extremely until the fatal outcome. Elevated T-helper levels in two patients from group A got normal during treatment. Hospital stay of group A patients is 4 days shorter compared to group B. Mortality in group A is 0%, in group B – 9,68%. In 2021, we conducted clinical observation of 8 pregnant women aged 25-38 years at 33-36 weeks of gestation, hospitalized for COVID-19. Lung CT showed pneumonia in all women. Treatment options during pregnancy are limited as many of medications for COVID-19 are contraindicated in pregnancy. All patients were treated with combination of two antibiotics and oxygen. Daily dose of anticoagulants depended on D-dimer values. All patients received probiotic formula, containing Lactobacillus bulgaricus DWT1 at daily dose of 30g, vitamins, pregnancy supplements. Only one patient received a small dose of corticosteroid for two days to improve fetal maturity. The general condition and paraclinical parameters of all improved on the 3^rd-4^th day. Inpatient treatment lasted up to 28 days. Delivery was performed by sectio caesarea. Women delivered live newborns with smooth uncomplicated postoperative period. Newborns are healthy, in good general condition with negative PCR tests.

The probiotic, containing Lactobacillus bulgaricus DWT1, modulates cellular immunity in COVID-19 patients – increases populations of NK-cells and cytotoxic T-lymphocytes and regulates T-helpers. Administration of this probiotic reduces COVID-19 death rate, shortens hospital stay and reduces the burden on healthcare system. This probiotic included in therapy successfully controls viral infection in pregnant women who give birth without complications to healthy live newborns not infected with COVID-19. The clinical observation results and the probiotic formula are subject of Bulgarian Patent №4294U and open new perspectives for application of Lactobacillus bulgaricus DWT1 as probiotic immunotherapy.

KEY WORDS:

Immunity; Lactobacillus Bulgaricus DWT1; Pregnancy; Probiotic; SARS-CoV-2

Scientific research of Laktera continues! Laktera is included in the only project in the field of medicine out of a total of 9 scientific projects that will be implemented during the 31st National Antarctic Expedition, led by Prof. Christo Pimpirev.

The probiotic foods Laktera have also been used under extreme conditions in the past. In the 1990s, with the permission of the General Staff of the Bulgarian Army, a study was carried out by a team from the Military Medical Academy, led by Prof. Alexandrov, among 56 Air Force pilots. Probiotic food, containing L. bulgaricus has been found to reduce total cholesterol and bad cholesterol, reduce triglycerides and increase good cholesterol, regulate blood sugar and humoral immunity. In 1996, a team from MMA, Sofia Medical University and Würzburg Medical University in Germany, under the leadership of Prof. Alexandrov, conducted a study of the effects of a probiotic food with L. Bulgaricus in the conditions of a simulated shipwreck near Shabla with 56 volunteers divided into two groups. The following beneficial effects were found in the volunteers who took the probiotic food – maintaining the body weight, arm muscle strength, maintaining the electrolyte balance and blood sugar levels. The probiotics Laktera have been used by the Bulgarian Army during the missions in Cambodia and Iraq.

On December 27, 2022, Laktera set sail for Antarctica. During the voyage, the members of the crew of the first Bulgarian vessel RV 421 Ss. Cyril and Methodius will take Laktera daily to study the effect of Lactobacillus bulgaricus DWT1 of water origin on the human health in these extreme conditions. The duration of the voyage is about 150 days.

The project studying the effects of Laktera is carried out by Nikola Vaptsarov Naval Academy together with MU “Prof. Dr. Paraskev Stoyanov„ and SU „Sv. Kliment Ohridski“. The topic of the project is „Study of the immunostimulating and protective effect of the probiotic foods, containing Lactobacillus bulgaricus in extreme stress situations during the upcoming trip, stay and return of our expedition to Antarctica on the Livingston Island“. The main objectives of the project are:

• To find out the therapeutic potential of Bulgarian probiotics, containing Lactobacillus bulgaricus for increasing the functional capabilities and the immune status of the body in volunteers, subjected to stress.
• To establish the potential of the probiotic food to serve as a protective factor or a kind of biological antidote in people placed in the conditions of an extreme physical and mental state.

The subject of the study – 20 sailors on the board of the vessel of Nikola Vaptsarov Naval Academy. They are divided into two groups of 10 people. The first group will take probiotic food Laktera at a dose of 15 g per day during the entire voyage and stay in Antarctica. The second group will not take Laktera.

All the participants in the research project will be tested for various biometrics: heart rate, breathing rate, blood oxygen, electrocardiogram, blood pressure, glycose levels, lipids, immune status, weight, bone mass, body fat, muscle mass, body fluids, body temperature, etc.

Also, the state of anxiety, depression and emotions, sleep quality and “burn out” syndrome will be examined by various psychological tests.

This project is expected to establish the effect of the probiotics Laktera on the human body for stimulating and maintaining the vital indicators when the body is subjected to extreme impact and stress. It is also expected to prove the efficacy of the Bulgarian probiotics, containing the first in the world strain of water origin Lactobacillus bulgaricus DWT1 under stress and social isolation.

Kosher certificate

The probiotics from Laktera family now have two types of Kosher Certificate – dairy and parve!

Certificate for ISO 22 000-2018 and GMP

For another year our company “Daflorn” – producer of the first probiotic of water origin in the world – successfully passed the audit by MG Global and received the highest level of international certification for food – ISO 22 000-2018 and GMP!

This Certificate is a guarantee for reliability and responsibility to all our partners and clients. Certification would not be possible without our new modern production facility, meeting all global requirements for food safety.

We will continue to create Laktera probiotics in our new home with care for your health and comfort!

Halal certificate

The probiotics from the Laktera family already have Halal Certificate issued by the Grand Mufti of the Muslim Denomination in Bulgaria!
Muslims are concerned whether the food they eat meets the Islamic religious tenets and traditions, humanitarian requirements, hygiene criteria and production methods. Halal certification is not just a marketing model for foods and food supplements, but proof that they are high quality and healthy. The Halal Certificate of Laktera family probiotics is valid all over the world.

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.