Mountain Food and Natural Probiotic in Nutrition

Study Overview
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Brief Summary
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Background/Objectives: The research discusses the importance of mountain foods/products and services in the context of healthier nutrition and behavior with the application to agribusiness. The purpose of the research is to highlight the necessity of human behavior in consuming mountain products as natural probiotics. Subjects/methods: As evidence, the research analyses mountain food and the associated entrepreneurship for Austria and other Central and Eastern European countries (foods/products and services), especially from the mountain area. The authors realized experimental research regarding representative mountain food from Central and Eastern European areas and used the Eurostat database. Data has been taken from Eurostat and processed in Excel and SPSS, using similar models of analysis from published research. Experimental analysis has been realized by the authors and collected from different recognized sources.

Detailed Description
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The study is focused on performing qualitative research for natural yogurt (animal resource), Allium sativum - garlic (plant resource), experimental analysis for natural honey (apicultural resource), namely honey-dew (manna) which is a mountain product from Maramureș, a region of Romania, and polyflora honey (Tilia - linden with mountain flowers) which is a hilly product from Moldova, a region of Romania, and clinical study on all the mountain products reminded before. After this study, it will be a comparative study of consumer behavior of mountain products from Central and Eastern European countries and Central and Southern states from the USA. This study is referring strictly to Central and Eastern European countries, especially Romania (but a country like Romania is representative of the other studied countries). The members of the family which will participate in the study will consume the probiotics reminded before. The family members, participants in the study, are consuming daily natural honey (one member of the family), yogurt (one member of the family), and garlic (two of them). After every 40 days, it will be verified, through laboratory analysis, the level of organism immunity. The analysis will be realized in Synevo Romania. It will be verified Biochemistry: Serum Calcium, Serum Magnesium, Serum Iron (Sideremia); Immunochemistry: Vitamin B12, Serum Folate, Vitamin 25-OH; Hematology: Leukocyte blood count with Hb, Ht, and indices (Leukocyte count, Erythrocyte count, Hemoglobin, Hematocrit, Mean erythrocyte volume, Mean erythrocyte hemoglobin, Mean Hb / erythrocyte concentration, Erythrocyte distribution width, Platelet count, L platelet count platelet distribution, Neutrophil, Lymphocyte, Monocyte, Eosinophil, Basophil); Zinc in the blood. It will be verified if properly used mountain animal and vegetal resources can become powerful fighters as natural probiotics against various diseases, or to prevent different immunity problems. Fermented natural dairy products are an extremely important component in the diet of the CEE population, especially for the rural population. Dairy products are often used as natural probiotics, especially within CEE countries. And, the most used mountain product is yogurt (or derivate products similar wit it, such as kefir, ayran, and so on). Within CEE countries, the most important species isolated from lactic bacteria fermented or raw products of dairy origin, are Lactococcus lactis, Leuconostoc sp., and Enterococcus sp. Probiotic dairy products have been analyzed and shown that in addition to nutritional benefits, they positively influence the physiology of the human body. The best-known dairy products are ice cream, cheese, yogurt, milk enriched with strains of Acidophilus and Bifidus, ayran, kefir, and kumis. Dairy drinks (fermented or unfermented) are considered foods that provide probiotics. In the fermentation process, lactic, acetic, and citric acid are naturally obtained, acids that are frequently used in the food industry to improve the organoleptic qualities of several products intended for consumption. The father of the paradigm according to which yogurt is one of the most powerful probiotics, especially for the digestive system, was Elie Metchnikoff, Nobel Laureate for Medicine. He postulated in the famous public presentation Old Age, held at the Society of French Agriculturalists in Paris on June 8, 1904, that the dependence between lactic acid bacteria and food makes it possible to apply methods to modify the intestinal flora by replacing bad bacteria with beneficial ones. He suggested that yogurt bacteria prevent and annihilate intestinal bacterial infections and hypothesize that regular yogurt consumption prolongs life. Lactic acid bacteria produce microbial metabolism, and the effect is a decrease in intestinal pH, which stops the proliferation of various proteolytic bacterial species. In this way, Metchnikoff kicks off the popularization of yogurt in Europe as a functional food with health benefits. Regarding plant resources, CEE countries are specialized in using different plants as natural probiotics. But, the most used plant resource is Allium sativum (garlic). Garlic consumption, as a natural probiotic, is just as old in the CEE as in many other countries around the world. Garlic is listed in the Dietary Guide for Americans 2015-2020. Garlic intake variants (capsules, aqueous extract, old alcoholic extract, etc.) are not very popular in the CEE, the preference being clear for the raw version. The experimental analysis of the study, realized by the authors in accordance with Romanian Research-Development National Institute for Apiculture - Bulletin analysis 39/29.01.2021, show that both types of natural honey have the same diastase index, meaning 29.4±0.18 unities on Goethe; mountain honey has sucrose (β-D-Fructofuranosyl α-D-glucopyranoside) of 2.41±0.43 %, g/g, while hilly products have 2.19±0.39 %, g/g; reducing sugar is in percent of 76.18±3.40 %, g/g and to the other of 75.67±3.37 %, g/g; 5-Hydroxymethylfurfural has a value of 0.10±0.01 mg/100g of honey for the mountain product and 0.19±0.03; mg/100g of honey for the hilly product; mountain honey has a superior PH value of 3.00±0.46 milliliters Sodium hydroxide solutions 1N/100 g of honey and the other of 2.80±0.43 milliliters Sodium hydroxide solutions 1N/100 g of honey; moisture contents has a percentage of 17.20±0.24 %, g/g for the mountain product and of 16.80±0.24 %, g/g for the other type of honey; pollen analysis present percentages of 98% of manna and 2% of mountain polyflora for the mountain product and of 95% of linden and 5% of manna for the other type of mountain; electrical conductivity present a value of 0.55 mS/cm, while ashes are in the value of 0,21±0,02 %, g/g (mountain product) and 0,44±0,03 %, g/g (hilly product); fructose (3S,4R,5R)-1,3,4,5,6-Pentahydroxyhexan-2-one) present a percentage of 41.06 %, g/g (mountain honey) and 40.10 %, g/g (hilly honey) and glucose (2R,3S,4R,5R)-2,3,4,5,6-Pentahydroxyhexanal) a percentage of 35.12 %, g/g (mountain product) and 35.57 %, g/g (hilly product). The physicochemical analysis shows that mountain honey has more qualitative values than hilly honey. Honey-dew (manna) contains minerals (calcium, magnesium, iron, potassium, phosphorus, selenium), antioxidants, organic acids, bioflavonoids, vitamins (especially C and group B), inhibin (strong bactericide), and enzymes. Minerals, proteins, and some acids are found in amounts five to ten times higher in mountain honey than in hilly honey. It has been verified if properly used mountain animals and vegetal resources (as natural probiotics) can prevent or fight for human immunity. Regarding Yogurt consumption, the influence on human immunity presents considerable values for magnesium, vitamin D, leukocyte, mean corpuscular hemoglobin, average hemoglobin/erythrocyte concentration, lymphocyte, monocyte, eosinophilic, basophil, and zinc. Different studies confirm the positive influence of yogurt consumption on immunity, as Yang et al. (2022), Ravindran et al. (2022), Kim et al. (2020), Gasparri et al. (2019), Meydani (2000), Leonard (2018), Jafari et al. (2016), Ajibola et al. (2012), Asemi et al. (2012). Allium sativum (garlic) influence positively the immunity of the organism, modifying the values of vitamin D, erythrocyte distribution width, platelets, and platelet distribution width, neutrophil, monocyte, and zinc. The influence of garlic on immunity is confirmed by numerous studies, such as Bandyopadhyay et al. (2021), Singh et al. (2021), Eor et al. (2020), Lee et al. (2020), Champagne et al. (2018), Divya et. al (2017), Percival (2016), Gholipour Kanani et al. (2014), Ried et al. (2014), Hamlaoui-Gasmi et al. (2012), Samson et al. (2012), Mukherjee et al. (2006), Hodge et al. (2002), Oluwole (2001), etc. Natural honey, especially manna honey, is considered to be one of the most powerful fighters of human immunity. According to our clinical analysis mountain honey has a positive effect on the adjustment of calcium, magnesium, vitamin B12, folates, vitamin D, mean corpuscular hemoglobin (MCH), Average Hgb / erythrocyte concentration, erythrocyte distribution width, platelets, lymphocyte, monocyte and eosinophilic. Complex research, clinical and experimental, sustain the importance of natural honey in immunodeficiency, studies as Attia et al. (2022), Bakour et al. (2022), Martínez-Puc et al. (2022), Ismail et al. (2022), Abdulrhman (2018), Sell et al. (2012), Ahmed et al. (2011), Al-Waili (2003), Kitzes (1943).

Official Title
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Mountain Food, as a Natural Probiotic. Evidence From Central and Eastern European Nutrition and Behavior

Conditions
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Immune Deficiency

Intervention / Treatment
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* Combination Product: Natural probiotics


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: members of the same extended family Exclusion Criteria: outside of the family

Ages Eligible for Study
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Minimum Age: 25 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Probiotic 1<br>Four members of an extended family: 1 white man of 71 years, 1 white woman of 75 years, 1 white man of 44 years, and 1 white woman of 44 years. | Combination Product: Natural probiotics<br>* Two of the group members use daily Allium Sativum - garlic, one use Natural honey, and 1 Yogurt. It is verifying the immunity of the organism after 40 days of using natural probitotics.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Laboratory blood tests 1 | Serum Calcium | September 2021 - April 2022 | 
| Laboratory blood tests 2 | Serum Magnesium | September 2021 - April 2022 | 
| Laboratory blood tests 3 | Serum Iron (Sideremia) | September 2021 - April 2022 | 
| Laboratory blood tests 4 | Vitamin B12 | September 2021 - April 2022 | 
| Laboratory blood tests 5 | Serum Folate | September 2021 - April 2022 | 
| Laboratory blood tests 6 | Vitamin 25-OH | September 2021 - April 2022 | 
| Laboratory blood tests 7 | Leukocyte count | September 2021 - April 2022 | 
| Laboratory blood tests 8 | Erythrocyte count | September 2021 - April 2022 | 
| Laboratory blood tests 9 | Hemoglobin | September 2021 - April 2022 | 
| Laboratory blood tests 10 | Hematocrit | September 2021 - April 2022 | 
| Laboratory blood tests 11 | Mean erythrocyte volume | September 2021 - April 2022 | 
| Laboratory blood tests 12 | Mean erythrocyte hemoglobin | September 2021 - April 2022 | 
| Laboratory blood tests 13 | Mean Hb / erythrocyte concentration | September 2021 - April 2022 | 
| Laboratory blood tests 14 | Erythrocyte distribution width | September 2021 - April 2022 | 
| Laboratory blood tests 15 | Platelet count | September 2021 - April 2022 | 
| Laboratory blood tests 16 | L platelet count platelet distribution | September 2021 - April 2022 | 
| Laboratory blood tests 17 | Neutrophil | September 2021 - April 2022 | 
| Laboratory blood tests 18 | Lymphocyte | September 2021 - April 2022 | 
| Laboratory blood tests 19 | Monocyte | September 2021 - April 2022 | 
| Laboratory blood tests 20 | Eosinophil | September 2021 - April 2022 | 
| Laboratory blood tests 21 | Basophil | September 2021 - April 2022 | 
| Laboratory blood tests 22 | Zinc in the blood | September 2021 - April 2022 |