# Screening of Some Romanian Raw Honeys and Their Probiotic Potential Evaluation
###### Abstract  
This study aimed to characterize raw honeys from different geographical origins in Romania, in respect of chemical composition, microbiological examination and evaluate their probiotic potential. The physico-chemical determinations were performed in APHIS-DIA Laboratory, Cluj-Napoca, Romania, using standard validated methods. Bacterial identification was performed for each sample and each colony type using Vitek\({}^{\circledR}\) 2 Compact 15 system and PCR amplification using 165 rDNA bacterial universal primers (27F, 1492R), species being confirm by sequences analysis. In five raw honey samples, we have identified probiotic bacteria, such as: _ Bacillus mycoides_, _ Bacillus thuringiensis_, _ Bacillus amploliquefaciens_, _ Bacillus subtilis_, and _ Bacillus velezensis_. Generally, all honey samples meet the standard values for chemical composition. However, one sample having 7.44% sucrose was found to have also probiotics bacteria from the genus Bacillus because sucrose is a substrate for probiotics development. In conclusion, the Romanian raw honey can be a potential reservoir of probiotics, which confer a health benefit for consumers. honeys, fructose oligosaccharides from honey; probiotic _ Bacillus_ strains; physicochemical characterization; antimicrobial activity +
Footnote †: journal: _Sci. 2021, 11_, 5816. [[https://doi.org/10.3390/app11135816](https://doi.org/10.3390/app11135816)]([https://doi.org/10.3390/app11135816](https://doi.org/10.3390/app11135816))
parts of plants or excretions of plant sucking insects on the living parts of plants which the bees collect, transform by combining with specific substances of their own, deposit, dehydrate, store and leave in honeycombs to ripen and mature.\" Its composition is rather variable and primarily depends on the botanical and geographical origin of the floral source, although certain external factors also play an important role, such as seasonal and environmental factors and its processing. Honey is a natural prebiotic composed of sugars, amino acids, enzymes, vitamins, and minerals [7]. Monosaccharides, glucose and fructose, are the major sugars (nearly 75%) and disaccharides, sucrose, maltose, turnanose, isomaltose, and maltulose, are in the small amount in honey [8]. Non-digestible carbohydrates (oligosaccharides and polysaccharides), some peptides and proteins, and certain lipids (esters and ethers) are considered prebiotics [9]. Prebiotics are non-digestible food ingredients that beneficially affect the host by selectively stimulating the growth and/or activity of one or a limited number of probiotic bacteria [10; 11; 12; 13; 14]. Fructooligosaccharides (FOS) are non-digestible carbohydrates, composed mainly of chains of fructose units with a terminal glucose molecule, such as fructans, oligofructans, glucofructans, inulins, or oligosaccharides, such as sucrose, inulobiose, and levanobiose [9; 15]. Sucrose is a fructooligosaccharide, which is prebiotic, and prebiotics are substrates for develop probiotics. If the concentration in fructooligosaccharides is higher, the probiotics are developing fast. High sucrose values in honeys are related to its botanical origin, for example, the honey dew or different honey maturity stages, or high nectar flux or artificial feeding of honey bees with sugar syrup [16]. Honeybees' relationship with probiotic microorganisms starts from the early metamorphosis stage of the larvae (day 4), when the feed is changed to honey, replacing royal jelly, until day 21 when adult larvae is ready to eclosionate. In this process, the larvae are immunologically stimulated due to probiotics contained in honey [2; 17]. For a long time, researchers believed that the source of lactic acid bacteria in honey was pollen and secretions of flowers that arrived in honey transported by honey bees. However, later studies proved that the lactic acid bacteria are present in the stomach of the honeybees; therefore, the source of lactic acid bacteria is the bee itself [7].