BG4294U1 - Biologically active product - Google Patents

Abstract

The utility model relates to a biologically active product for nutritional support of human immunological disorders, comprising (in weight %): goat's milk with fat content from 0.1% to 30% - from 98.0 to 98.8; active cells of the microorganism strains Lactobacillus delbrueckii ssp. bulgaricus DWT1 CCM No. 7992, Streptococcus thermophilus DWT4 CCM No. 7992, Streptococcus thermophilus DWT5 CCM No. 7993, Streptococcus thermophilus DWT6 CCM No. 7994, Streptococcus thermophilus DWT7 CCM No. 7995 and Streptococcus thermophilus DWT8 CCM No. 7996 in a ratio 1:0. 2:0.2:0.2:0.2:0.2 - from 0.1 to 0.2 and fructooligosaccharides with a degree of polymerization of 95% - from 1.0 to 2.0. The application of the biologically active product as a dietary probiotic supplement accompanying the complex treatment regimen in patients with COVID-19 leads to successful control of the viral infection and has promising qualities of the food of the future for fighting viral infections.

Description

Field of technique

The utility model relates to a biologically active product with probiotic properties applicable as a nutritional support agent for human immunological disorders.

Prior art

It is known that products obtained by lactic acid fermentation have been produced for centuries. They are a healthy food that most people take daily from early childhood. For the preparation of these foods, various lactic acid microorganisms are used both alone and in combination, including Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus acidophilus, Streptococcus thermophilus and others with proven probiotic properties.

It is also known that Lactobacillus delbrueckii subsp. bulgaricus is a specific type of microorganism, significantly different from other probiotic bacteria. It is believed that this bacterium under natural conditions develops only in a certain geographical area, coinciding with the territory of Bulgaria and corresponding to a large extent to the territory of ancient Thrace. Outside this geographical area, the microorganism mutates and does not develop. It is also known that Lactobacillus delbrueckii subsp. bulgaricus can cooperate only with Streptococcus thermophilus in lactic acid fermentation in humans and animals. In this unique cooperation, they mutually support each other, which leads to a multiple strengthening of the beneficial and balancing of their side effects. These beneficial effects include a positive effect on the natural human microflora, competitive exclusion of pathogens and stimulation/modulation of mucosal immunity, and the knowledge of this has been used in a number of developments to obtain probiotic products. For example, a product containing selected probiotic strains of Lactobacillus bulgaricus and Streptococcus thermophilus was found to suppress the proliferation of human colon carcinoma cell lines HT-29 in a dose-dependent manner [1]; Lactobacillus bulgaricus DWT1 strain in particular was also found to have hepatoprotective activity against paracetamol-induced liver injury in Wistar rats [2], and a probiotic formulation containing Lactobacillus bulgaricus DWT1 and Streptococcus thermophilus DWT4 strains repolarized macrophages from the M2 to M1 state with the potential to development of newer strategies to intervene in tumor growth [3].

All these publications, based on detailed studies, point to a possibility of application of probiotic microorganisms for the development of means for maintaining the normal immunity of man, for nutritional support of his immunity and as means for probiotic immunotherapy.

Technical nature of the utility model

The task of the present utility model is to offer a biologically active product combining the beneficial properties of specially selected probiotic strains of lactic acid microorganisms to provide nutritional support for human immunological disorders.

In its essence, the useful model is a biologically active product with probiotic properties, representing a combination of milk or milk ingredients with valuable metabolites from the vital activity of specially selected strains of lactic acid bacteria.

In particular, the biologically active product, according to the useful model, includes the following components in their quantitative proportions (in % by weight):

- goat milk with fat content from 0.1% to 30% - from 98.0 to 98.8;

- active cells of the strains of microorganisms Lactobacillus delbrueckii ssp. bulgaricus DWT1 CCM No. 7992, Streptococcus thermophilus DWT4 CCM No. 7992, Streptococcus thermophilus DWT5 CCM No. 7993, Streptococcus thermophilus DWT6 CCM No. 7994, Streptococcus thermophilus DWT7 CCM No. 7995 and

Streptococcus thermophilus DWT8 CCM No. 7996 in a ratio of 1:0.2:0.2:0.2:0.2:0.2 - from 0.1 to 0.2;

- fructooligosaccharides with a degree of polymerization of 95% - from 1.0 to 2.0.

Goat's milk, included in the composition, according to the useful model, is a very good source of calcium and the amino acid tryptophan. It is a good source of proteins, phosphorus, riboflavin (vitamin B2) and potassium, which is why it is both a suitable raw material for the multiplication of selected, according to the beneficial model, probiotic microorganisms, and as a component in the composition of the biologically active product.

In turn, the fructooligosaccharides included in the biologically active product are short-chain, low-energy carbohydrates (1.5 Kcal/g) that belong to the group of indigestible oligosaccharides. They are obtained through bioenzymatic reactions from onion, garlic, burdock and banana plants. They are characterized by a low molecular mass and a content of different numbers of monosaccharide units in their molecules. They have a polymerization degree of 95% with a polymer length of 3 to 5 monomers, of which: glucose-2(fructose) - 37%, glucose-3(fructose) - 53% and glucose-4(fructose) - 10%.

Fructooligosaccharides appear as ballast substances (dietary fiber), which implies their strong bifidogenic effect. They influence the physiological functions of digestion by improving the pH level in the stomach and colon and improving hepatic lipogenesis, i.e. they assist in the reformation of carbohydrates into triglycerides, thereby supporting lipid metabolism and the maintenance of homeostasis, which predetermines their huge physiological role for the implementation of important processes in the body - they have a supportive, healing and protective effect on the gastrointestinal microflora. These properties and qualities of fructooligosaccharides make them a suitable component in a biologically active agent for modulating human immunity.

The association of six specially selected strains of probiotic microorganisms, each of which has been deposited in the Czech Microorganism Depository Bank for the purpose of the patent procedure, according to the Budapest Treaty (BTM), are part of the isolated for the first time in the world by N. Aleksandrov and D. Petrova strains of lactic acid microorganisms from pure spring water in Stara planina, Bulgaria [4]. They have extremely effective qualities and properties that make them suitable for inclusion as an active component in the composition of the biologically active agent, according to the useful model, namely: proven good antimicrobial activity, including a remarkable bacteriostatic effect and a moderate bactericidal effect; proven safety for human consumption and very good survival and resistance in extreme conditions of the human stomach and intestine; proven high viability during long-term storage at room temperature, as well as proven high survival after lyophilization.

The components included in the composition - goat's milk and fructo-oligosaccharides allow microorganisms to multiply together, which leads to obtaining a product extremely rich in living microorganisms, the quality of which is especially valuable for modulating human immunity through healthy nutrition.

The utility model is illustrated by the following exemplary embodiments without limiting its scope.

Example 1. According to the present example, the biologically active product comprises (g):

- 10% aqueous solution of dry standardized full-fat or skimmed goat's milk - 5,000 g;

- dry starter culture, including active cells of the strains of microorganisms Lactobacillus delbrueckii ssp. bulgaricus DWT1 CCM #7992, Streptococcus thermophilus DWT4 CCM #7992, Streptococcus thermophilus DWT5 CCM #7993, Streptococcus thermophilus DWT6 CCM #7994, Streptococcus thermophilus DWT7 CCM #7995 and Streptococcus thermophilus DWT8 CCM #7996 in a ratio of 1:0.2:0.2:0.2: 0.2:0.2 5.0 g;

- fructooligosaccharides with degree of polymerization 95% - 50.0 g.

Preparation: To obtain the biologically active product, according to the useful model, a starter culture is first obtained from the selected strains of probiotic microorganisms, after which a 10% aqueous solution of dry standardized full-fat or skimmed goat's milk is subjected to sterilization at 95°С-96°С for 30 min, cooled to 45°С and neutralized with a 10% solution of sodium base until reaching pH 6.6-6.7. Next, a 1:1 aqueous solution of fructooligosaccharides is prepared, which is heated to boiling, allowed to cool, and added to the aqueous milk solution in a 10:1 milk:fructooligosaccharides ratio.

The starter culture obtained as above is added to the mixture thus obtained, followed by fermentation for 12 h at 43°C-45°C and the resulting product is subjected to lyophilization. The product thus obtained is stored at room temperature.

Example 2. Effectiveness of the biologically active product, according to the utility model, to affect cellular immunity in patients with COVID-19.

Resistance to viral diseases depends mainly on the activity of the immune response of the human body. The main lymphocyte populations of cellular immunity are T-cytotoxic lymphocytes, T-helpers, NK cells and B lymphocytes. NK cells are the "natural killers" that exhibit spontaneous cytotoxicity against virally infected cells and tumor cells. T-cytotoxic lymphocytes have a cytotoxic effect and directly destroy genetically foreign cells (cells infected with viruses, bacteria or tumor cells). T-helpers have an auxiliary function - they produce cytokinins and support cells of innate immunity - macrophages and cells of cellular immunity - T-cytotoxic lymphocytes and B-lymphocytes. B-lymphocytes are responsible for humoral immunity by mediating the production of antigen-specific immunoglobulins and provide immunological memory upon re-encounter with the antigen.

SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2) is a strain of the coronavirus that causes the pandemic disease COVID-19. The disease affects the immune system but also changes the gut microbiota [5-7]. COVID-19 is inhaled and most often damages the microbiome of the respiratory tract. The disease is more severe in patients with accompanying diseases, among which the most common are arterial hypertension, cardiovascular and cerebrovascular diseases, diabetes mellitus and metabolic syndrome. The treatment of COVID-19 requires an interdisciplinary approach [8]. In addition, pregnant patients are much more severely affected by COVID-19 due to the lack of drugs that can be used during pregnancy - SARS-CoV-2 compromises placental antibody transfer [9]. For these reasons, non-traditional methods such as monoclonal antibodies are more often used to treat patients with COVID-19 [10], and for prevention as well as for rehabilitation, people turn to a healthy lifestyle and a balanced diet, so that they can to exercise some control over their immune system [11-12]. In this aspect, the biologically active product, according to the utility model, is suitable as a means of nutritional support of immunity in COVID-19.

The study of the effect of the biologically active product containing the probiotic strains Lactobacillus delbrueckii ssp. bulgaricus DWT1 CCM No. 7992, Streptococcus thermophilus DWT4 CCM No. 7992, Streptococcus thermophilus DWT5 CCM No. 7993, Streptococcus thermophilus DWT6 CCM No. 7994, Streptococcus thermophilus DWT7 CCM No. 7995 and Streptococcus thermophilus DWT8 CCM No. 7996 as agents for modulating cellular immunity in patients with COVID-19, was performed on three types of sites: inpatients with COVID-19; ambulatory patients with COVID-19 and pregnant with COVID-19.

1. Hospitalized patients with COVID-19

The clinical study was conducted on 78 patients hospitalized in the period from 29.04.2020 to 16.12.2020 in the University General Hospital for Active Treatment "Dr. G. Stranski" EAD Pleven with Dg: U07.1 2019-nCoV. The study was a prospective, cohort, randomized, open-label study.

Statistics were done by IBM SPSS version 23. Patients gave informed consent.

For the purpose of the study, patients were randomly divided into two groups: Group A - 47 patients received the biologically active product, according to the useful model, in an amount of 15 to 45 g depending on the severity of the clinical picture as part of a complex treatment scheme for COVID -19 and Group B - 31 patients who received a standard treatment regimen for COVID-19 without receiving a biologically active product.

From group A, 31 patients (65.96%) were male and 16 (34.04%) were female. The patients were aged from 16 to 88 years and were distributed as follows: from 16 to 35 years - 4 (8.51%); from 35 to 55 years - 15 (31.91%); from 55 to 65 years - 18 (38.30%) and over 65 years - 10 (21.28%).

From group A, 29 patients (61.70%) had comorbidities, with 18 (38.30%) of them having more than one comorbidity. The most common accompanying diseases are: arterial hypertension and cerebral and cardiovascular disease in 25 of the patients (53.19%); diabetes mellitus and metabolic syndrome in 17 patients (36.17%); two of the patients (4.26%) had carcinoma.

From group B, 20 patients (64.52%) were male and 11 (35.48%) were female. The patients were aged from 20 to 82 years and were distributed as follows: from 20 to 35 years - 2 (6.45%); from 35 to 55 years - 8 (25.81%); from 55 to 65 years - 13 (41.93%) and over 65 years - 8 (25.81%).

From group B, 21 patients (67.74%) had comorbidities, with 13 (41.94%) of them having more than one comorbidity. The most common accompanying diseases are: arterial hypertension and cerebral and cardiovascular disease in 17 of the patients (54.84%); diabetes mellitus and metabolic syndrome in 12 patients (38.71%); one patient (3.23%) had carcinoma.

Obviously, the two groups of patients, A and B, do not differ significantly in gender and age distribution, frequency and type of accompanying diseases.

All 78 patients from group A and group B were hospitalized with a positive PCR test for COVID-19. The most common data from the anamnesis upon admission are: persistent fever up to 39-39.5°C, which is not affected by antipyretics, cough with sputum, shortness of breath, astheno-adynamic syndrome, loss of smell. All patients in both groups had Rogr of lung and heart performed on admission.

Ro gr showed pneumonia in 43 (91.49%) of patients in group A and bronchitic changes in 4 (8.51%) of them.

Ro gr showed pneumonia in 28 (90.32%) of patients in group B and bronchitic changes in 3 (9.68%) of them.

The data showed that there were no significant differences in radiographic findings between group A patients and group B patients.

X2=0.031, df=1p=0.86 => no statistical difference in distribution according to X-ray findings.

On admission, all patients underwent paraclinical blood tests as follows: CK (er, Hb, MCV, Hct, Leuc, Thr, Sg, Mo, Ly), ESR, glucose, fibrinogen, total protein, albumin, lactate dehydrogenase (LDH), troponin, prothrombin index, prothrombin time in %, prothrombin time INR, D-dimer, aspartate aminotransferase (ASAT), alanine aminotransferase (ALAT), gammaglutamyltransferase (GGT), total bilirubin, direct bilirubin, alkaline phosphatase, CRP, serum iron, TSHK, creatinine, urea. They were followed during the hospitalization period. The acid-base status of CAS was monitored multiple times daily for partial pressure of O2 (pO2) and partial pressure of CO2 (pCO2) and oxygen saturation.

Blood sugar is monitored multiple times daily in patients with diabetes mellitus.

The most common deviations in the examined blood parameters of the patients at admission make an impression on the expressed lymphopenia with granulocytosis, a strong increase in LDH, elevated D-dimer in some of the patients, a strong decrease in pO ₂ and impaired saturation. The results were similar in both patient groups A and B.

The following lymphocyte subpopulations of cellular immunity have been traced from immunological paraclinical studies by immunophenotyping of leukocytes: total T-lymphocytes (CD3 marker); T-helpers (CD4 marker); T-cytotoxic lymphocytes (CD8 marker); Th/Tc ratio; activated T lymphocytes CD3+ HLA- DR; T-helpers CD3+ CD4+ CD8-; T-cytotoxic suppressors CD3+ CD4- CD8+; Blymphocytes (CD19 marker); peripheral NK cells (CD3-/CD16+ CD56+).

From the indicators of humoral immunity, the following were followed: total immunoglobulins, IgM, IgG, IgA, immunoglobulin E in 30 patients from group A and 21 patients from group B.

Study results: All 47 patients in group A who received the biologically active product were discharged with a negative PCR test. At discharge, a control Ro gr of lungs and heart was done. X-ray examination shows that in 26 patients (55.32%) there is no pneumonia, and in the remaining 21 (44.68%) the pneumonia is in reverse development.

From the paraclinical blood tests performed at discharge, discrete lymphopenia was detected in only 3 of the patients (6.38%), while the remaining patients had normal values of lymphocytes. Granulocytosis in control studies was found in 2 of the patients (4.26%). LDH values remained elevated, but extreme elevation of LDH over 200% was found in only 2 patients (4.26%), and moderate elevation of LDH up to 200% in 13 patients (27.66%). D-dimer in all is normal. All patients were discharged with oxygen saturation above 90%.

The results for the cellular immunity activity at discharge of the patients in group A who received the biologically active product as part of the complex treatment therapy for COVID-19 are presented in Table 1 below.

Table 1.

Values Inde*^^ Reference values in number of petton is (%) Increased values En number of patients (%) Decreased values in number of patients (%) Total T lymphocytes 24 /51.0W 19/40,42/ 4/3.52/ T helpers 39 /82.97/ 6/17,03/ Cytotoxic T lymphocytes 14/29.79/ 32/86,09/ 1 /2,12/ Β lymphocytes 44 /93.62/ 3/6,38/ Total NK calls 2/4.28/ 45 /95.74/

BG 4294 UI

None of the patients in group A required intubation, and no patients died (including patients with carcinoma as a comorbidity).

Of all 31 patients in group B, 28 were discharged with a negative PCR test and three died. X-ray examination at discharge showed that in 11 patients (35.48%) there was no pneumonia, and in 17 patients (54.83%) the pneumonia was in reverse development.

From the paraclinical tests performed at discharge, lymphopenia was found in 10 of the patients (32.26%), while in the remaining patients the lymphocyte values were at the lower limit of the norm. Follow-up studies revealed granulocytosis in 9 patients (29.03%). LDH values remained elevated, as an extreme increase of LDH above 200% was found in 8 patients (25.81%), and a moderate increase of LDH up to 200% - in 17 patients (54.84%). D-dimer was normal in all 28 discharged patients (90.32%). Oxygen saturation was above 90% in all 28 patients at discharge.

The results of the cellular immunity activity of all 31 patients in group B who did not receive the biologically active product, according to the utility model, as part of the complex treatment regimen for COVID-19, are presented in Table 2 below.

Table 2.

Go to them Index Reference values In number of patients (%) Increased values in number of patients (%) Decreased veki&s in number of patients (%J Total T lymphocytes 14/45.16/ 5/16.13/ 12 /36.71/ T helpers 15/+8.38/ 3/9.68/ 13/41.94/ Cytotoxic T lymphocytes 17/54,34/ 5/10.35/ 8/25,81/ In lymphocytes 28 /90,32/ 1 /3.23/ 2/6.45/ Total NK cells 7 /22.58/ 22/70.97/ 2/6.45/

BG 4294 UI

4 of the patients (12.90%) from group B required intubation and were transferred to the intensive care unit. They stayed an additional 15 to 30 days, and 3 of them (9.68%) died. One of the deceased had an accompanying disease - colon carcinoma. The other two have accompanying diseases - arterial hypertension, diabetes mellitus and cardiovascular disease. On admission, two of the decedents had elevated values of Helper lymphocyte populations and decreased values of total NK cells, T-cytotoxic lymphocytes, and total T lymphocytes. At exit there was a significant increase in T-helper cells and retention of low levels of NK cells and T-cytotoxic lymphocytes compared to levels at hospitalization.

2. Outpatients treated with COVID-19

In the period from June 2020 to April 2021, doctors from MC Eurohealth-Bulgaria Ltd. are monitoring 54 patients with COVID-19. During active infection with COVID-19, these patients are consulted by phone or online. All patients were positive for COVID-19, with 10 (18.52%) of them being diagnosed with a PCR test, and the remaining 44 (81.48%) with a rapid coronavirus antigen test to determine the presence of viral antigens in the patient's nasopharynx. All patients were treated at home.

(66.67%) of the observed patients were male and 18 (33.33%) were female aged between 30 and 80 years. The most common accompanying diseases are arterial hypertension - in 38 (70.37%) patients, diabetes mellitus - in 6 (11.11%), of which 3 (5.56%) are on insulin. Three of the patients (5.56%) had neoplastic processes, and one of them, a 62-year-old man, had diffuse large cell Intracellular non-Hodgkin's lymphoma and was undergoing chemotherapy. Patients take a test for COVID-19 due to complaints of temperature up to 39.5°C, runny nose, sore throat, cough, astheno-adynamic syndrome. In 18 patients (33.33%), pneumonia was diagnosed by radiographic examination of the lungs and heart. At the start of treatment, only 11 of the patients (20.37%) had paraclinical blood tests, including PKK, ESR. It is impressive that 8 of the patients have marked lymphopenia. Patients with diabetes mellitus monitor blood sugar several times a day with a glucometer and once a day monitor acetone in the urine with a test strip. Patients with pneumonia monitor saturation at home with a pulse oximeter.

All patients were treated with antiplatelet agents (Clopidogrel), vitamins, antipyretics, nutrients and the biologically active product, according to the useful model, in a daily dose of 30 g. A quinolone antibiotic (Levofloxacin) was included in the therapy of 45 patients (83.33%). In 4 of the patients (7.40%), corticosteroids were included in the scheme. 27 of the patients (50%) were taking antihistamines (Claritin). All medications were taken orally.

Depending on the blood sugar values, the amount of insulin to be administered to type 2 diabetes patients with insulin depletion is determined by a doctor dynamically, over the phone, several times a day.

Patients with pneumonia monitor Og saturation with a pulse oximeter.

Patients with arterial hypertension monitor the RR values three times a day and, if necessary, the therapy for arterial hypertension is adjusted.

Due to a drop in saturation below 70% in a 79-year-old female patient with comorbidities of arterial hypertension and type 2 diabetes mellitus with insulin depletion, Og was required to be given by mask with an oxygen cylinder at a dose of 6 Ι/h at home for two days. The patient with non-Hodgkin's lymphoma temporarily (for 20 days) interrupts the chemotherapy treatment course.

Research results: By the end of the quarantine period (14 days), all patients recovered without complications. None of the observed 54 patients required hospitalization. After completion of the home treatment of COVID-19, the patients continued to take an oral antiplatelet agent (Clopidogrel) and the biologically active product, according to the utility model, at a daily dose of 15 g for another 3 months.

Patients with concomitant diseases continued to take their therapy for arterial hypertension and diabetes mellitus. The patient with non-Hodgkin's lymphoma continued the course of chemotherapy.

After completion of the course of treatment for COVID-19, tests for the activity of cellular immunity were performed on an outpatient basis in 21 of the 54 patients observed. The test results of these 21 patients (100%) are presented in Table 3 below.

Table 3.

values Index Reference values In number of patients (%) Increased values in number of patients (¾) Orcronsed values in number of patients (%) To'^l T lymptiocytea 9 /42,56/ 10/47,52/ 2/9.52/ T helpers $1 /75.19/ 5/23.80/ Cytdlcude t IVffiphocM®s 6 /28.67/ 15/71,42/ in Ivmonocvtea 19/50.48/ 2/9.52/ Total NK cells 2 /5.52/ 19/90.4"

BG 4294 UI

3. Pregnant women and COVID-19

The object of clinical observation are 8 pregnant women hospitalized in the period June 2020 - April 2021 in the Saint Anna University General Hospital for Active Treatment, Sofia, in the Obstetrics and Gynecology Clinic with a diagnosis of COVID-19.

The patients were aged between 25 and 38 years and all were pregnant with one child. 7 of them are at 3336 weeks of gestation and one of them is at 26 weeks of gestation. Before hospitalization, the 8 women were treated for COVID-19 on an outpatient basis at home for 3 to 5 days with symptomatic agents. Due to deterioration of the clinical condition - onset of shortness of breath, cough, high temperature 39-39.5°C, they were admitted for hospital treatment. The patient in the 26th week of gestation was brought for treatment from a hospital in another city. None of them have accompanying diseases. At hospitalization, all have a positive PCR test for COVID 19.

X-ray examination of the lung is contraindicated in pregnancy, which is why a computed tomography (CT) examination is performed in pregnant women. Lung CT showed pneumonia in all 8 women, with inflammatory changes affecting more than 20% of both lung halves. In the 38-year-old woman (elderly) inflammatory changes affect more than 30% of the lungs.

On admission, all patients underwent paraclinical blood tests as follows: PKC, ESR, glucose, fibrinogen, total protein, albumin, lactate dehydrogenase LDH, troponin, prothrombin index, prothrombin time in %, prothrombin time INR, D-dimer, AST, ALAT, GGT, total bilirubin, direct bilirubin, alkaline phosphatase, CRP, serum iron, TSH, creatinine, urea, serum magnesium level. They were followed during the hospitalization period. Acid-base status (AAC) was monitored multiple times daily for partial pressure of _O2 (pO2 ₎ and pCO2 and oxygen saturation.

D-dimer was monitored daily in all. Serum magnesium level was monitored twice a week.

Microbiological cultures of vaginal, nasal and throat secretions and urocultures are carried out in the monitored pregnant women every 5 days.

Fetal tones are monitored daily.

The reference values of the blood indicators of pregnant women differ from those of other patients. For example, the reference values of leukocytes (WBC) are from 3.5 to 10.5 G/l, but in pregnancy if the leukocytes are below 15 G/l, leukopenia is considered. Reference values of D-dimer are 100-500 ng/ml, and in pregnant women the norm is up to 1500 ng/ml.

The most frequent deviations from the studied blood indicators at admission are: lymphopenia, granulocytosis, leukopenia and increase of LDH and D-dimer. In the 38-year-old top woman, D-dimer reached 15,000 ng/ml. Oxygen saturation was reduced in all of them.

Treatment options for pregnant women are severely limited, as many of the medications to treat COVID-19 are contraindicated in pregnancy.

7 patients were treated with a double antibiotic combination, including Meronem (from the group of carbapenems) and Sulperazon (a cephalosporin antibiotic and Sulbactam), administered i.v., infusion of glucose-salt solutions, vitamin C and antipyretic paracetamol. Anticoagulant treatment is also dosed with daily monitoring of Ddimer. Clexan was administered as an antithrombotic medication in pregnant women in a dose of 2 x 60 mg (0.4 ml) to 1 x 60 mg (0.2 ml), depending on the D-dimer values and the severity of the clinical picture. Aspirin and Clopidogrel were not administered to pregnant patients. Oxygen saturation was improved with mask oxygen at a dose of 6 l/min.

The seven pregnant patients also take a biologically active product, according to the useful model, in a daily dose of 2 x 15 g. They also take the standard vitamins and supplements for normal pregnancy - vit. D, folic acid, Omega 3 fatty acids, vitamins and minerals for pregnant women. Of the minerals, the most important is the intake of magnesium in a daily dose of 800-1200 mg as a tocolytic, cardioprotector and means of preventing premature birth.

Due to a rapid deterioration of the general condition with very severe shortness of breath, the 38-year-old woman in the 34th week of gestation was transferred to an intensive care and resuscitation clinic 6 h after hospitalization. The treatment carried out there included the complex treatment scheme described above, with the anticoagulant Clexane in a maximum dose of 3 x 60 mg (0.6 ml) in the first few days. The biologically active product is now in a daily dose of 3 x 15 g. In addition, the patient took a corticosteroid (Dexamethasone 2 x 8 mg daily dose) only for 48 h in order to improve the maturity of the fetus. Treatment with hyperimmune plasma was also applied.

Research results: The general condition and paraclinical indicators of the seven women treated at the Obstetrics and Gynecology Clinic, who were additionally given nutritional therapy with a biologically active product, improved on the 3rd-4th day of hospitalization. The patient's D-dimer in the Intensive Care and Resuscitation Clinic dropped from 15,000 to 4,500 ng/ml as early as the 3rd day of hospitalization (the more severe the course of COVID-19 in pregnant women, the higher the D -dimera), and pO ₂ increased and no intubation was required.

Inpatient treatment of COVID-19 in the 8 pregnant patients lasted from 20 to 28 days until resorption of the inflammatory foci in the lung and a negative PCR test.

All patients, except the 38-year-old, who was voluntarily discharged after a negative PCR test in good general condition (subject to pregnancy follow-up and delivery in another city), continued to take standard pregnancy supplements and the biologically active product, according to the useful model, in a daily dose of 2 x 15 g. During the hospital stay, they were given birth by sectio caesarea, performed with spinal anesthesia. Women have a smooth, uncomplicated postoperative period, and a sharp improvement in their condition is observed after delivery. Of the 7 women with singleton pregnancies, 7 live births were born - all healthy and in good general condition. After birth, newborns are quarantined. All infants underwent PCR testing on postnatal days 1 and 8, and all PCR tests were negative.

Mothers and newborns were discharged in good general condition with negative PCR tests. Their home therapy also includes the biologically active product in a daily dose of 15 g for 3 months.

All the results of the conducted experiments on the effect of the biologically active product, according to the useful model, to modulate human immunity in the viral disease COVID-19, indisputably lead to the conclusion that the probiotic strains of microorganisms included in the product actively participate in the complex, not yet fully known, a cascade of immune responses from macrophages, T lymphocytes, cytokinins and occupy a central place in the orchestration of the immune response. The application of the biologically active product in a daily dose of 30-45 g as nutritional probiotic therapy, accompanying the complex treatment scheme in patients with COVID-19, leads to successful control of the viral infection and has the promising qualities of a food of the future to fight viral infections.

Claims (1)

Biologically active product for nutritional support in human immunological disorders with the participation of selected strains of probiotic microorganisms, characterized by the fact that it includes (in wt. %): - goat milk with a fat content of 0.1% to 30% - from 98.0 to 98.8; - active cells of the strains of microorganisms Lactobacillus delbrueckii ssp. bulgaricus DWT1 CCM #7992, Streptococcus thermophilus DWT4 CCM #7992, Streptococcus thermophilus DWT5 CCM #7993, Streptococcus thermophilus DWT6 CCM #7994, Streptococcus thermophilus DWT7 CCM #7995 and Streptococcus thermophilus DWT8 CCM #7996 in a ratio of 1:0.2:0.2:0.2: 0.2:0.2 - from 0.1 to 0.2; - fructooligosaccharides with a degree of polymerization of 95% - from 1.0 to 2.0