Effect of Natural Food on Gut Microbiome and Phospholipid Spectrum of Immune Cells in COVID-19 Patients

Study Overview
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Brief Summary
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The efficacy of natural foods such as freeze-dried mare's milk (Saumal) in post-COVID syndrome therapy has not been studied. The literature review has shown that researchers have focused more on evidence-based medications and less on natural products. Some raw foods, such as freeze-dried mare's milk, contribute to forming complete immune complexes and have antioxidant, membrane stabilizing, and antiviral effects. The use of Saumal proved its effectiveness in patients suffering from chronic hepatitis C. After 4 weeks of using freeze-dried mare's milk, the biodiversity of the intestinal microbiome was increased. The content of bacteria secreting short-chain fatty acids also increased. The study aims to confirm these effects at the gene level in patients who underwent COVID-19. This study will allow us to develop a highly evidence-based component of rehabilitation therapy in patients after COVID-19.

Detailed Description
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The study consists of three stages. Stage I. Randomized division of 76 patients under study into a main group of 38 patients receiving Saumal for 4 weeks and a control group of 38 patients not receiving Saumal. All respondents have suffered from COVID-19 in the last 6 months. At the moment of the research, all patients will be staying in rehabilitation centers in Almaty City. The study's biological material will be collected from the respondents (gut microbiome and antiphospholipid antibodies). Stage II. The main group will be given Saumal for four weeks, after which the gut microbiome and antiphospholipid antibodies will be collected from both groups once again. This will allow us to assess the impact of freeze-dried mare's milk on the condition of the gut microbiome and antiphospholipid antibodies. Stage III. Data analysis. Based on the results of gut microbiome sequencing and ELISA (antiphospholipid antibodies) in the main and control groups, we determine the dynamic changes of that parameters during COVID-19 rehabilitation. According to the results, the effectiveness of Saumal itself regarding these 2 parameters will be evaluated. Develop recommendations and an algorithm for patient management after COVID-19 to prevent complications.

Official Title
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Effect of Natural Food on Gut Microbiome and Phospholipid Spectrum of Immune Cells in COVID-19 Patients

Conditions
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COVID-19

Intervention / Treatment
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* Dietary Supplement: Freeze-dried Mare Milk (Saumal)


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Age 18 years and older; Patients in rehabilitation after COVID-19; Signed informed consent; Presence of patient's history of COVID-19, reliably established by PCR+ /presence of IgG/ diagnosis of coronavirus pneumonia on Computer Tomography based on discharge from hospital or outpatient records. Exclusion Criteria: Chronic inflammatory bowel disease; Gut microbiota transplantation; Chronic pancreatic disease, period of exacerbation; Liver cirrhosis, Metavir stage 3-4; Any disease in the decompensation stage; Neuralgic and psychological disorders that interfere with the study; Cancer; Non-transportable patients; Patients who do not reside in Almaty; Patients who have refused to participate in the study.

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 75 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Non-Randomized
Intervention Model: Parallel Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Main Group<br>Patients who had suffered from COVID-19 in the six months before the start of the study. Patients will receive Saumal (freeze-dried mare milk) for four weeks. | Dietary Supplement: Freeze-dried Mare Milk (Saumal)<br>* Freeze-dried Mare Milk (Saumal) is frequently reported for having therapeutic and dietary properties associated with specific chemical composition and certain physical properties of the product. It contains a total of about 40 biologically active components, the most important of them vitamins A, C, B1, B2, B6, and B12, amino acids, enzymes, and trace elements (low molecular weight peptides, lactalbumins, and globulins). Saumal will be administered to patients at a dose of 20 grams of dry powder per 200 ml of water, 2 times a day, 30 minutes before the meal, for 4 weeks.<br>|
| No Intervention: Control Group<br>Patients who had suffered from COVID-19 in the six months before the start of the study. There will be no intervention. | |

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Gut Microbiome | Intestinal microbiome studies will be performed using the Ion 16S™ Metagenomics Kit and Ion Torrent Next Generation Sequencing (NGS) technology. Creation of libraries for NGS sequencing. The commercial Ion 16S™ The Metagenomics Kit includes two primer pools that can amplify the hypervariable regions of 16S rRNA in bacteria. The first pool produces V2-4-8, while the second pool allows for the production of V3-6 and 7-9 regions of 16S rRNA.Sequencing will be performed on an Ion S5 analyzer using an Ion 520 Chip Kit. Once the PCR products have been run, it is important to follow standard procedures for fragment end recovery, adaptor ligation, nick gap repair, qualitative and quantitative assessment of non-amplified libraries, and pooling. | 4 weeks | 
| Antiphospholipid antibodies | IgG or IgM autoantibodies to cardiolipin, phosphatidyl serine, phosphatidyl-inositol, phosphatidyl acid, and β2-glycoprotein I are determined in human venous blood serum by enzyme immunoassay (ELISA) using the Anti-Phospholipid Screen IgG/IgM kit. | 4 weeks | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Immune status changes | Levels of immune status markers (Immunoglobulin G, Immunoglobulin M, Immunoglobulin A) will be detected from blood samples. | 4 weeks | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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COVID-19, gut microbiome, antiphospholipid antibodies, rehabilitation, food products