# Screening of Some Romanian Raw Honeys and Their Probiotic Potential Evaluation
## 4 Discussion  
This study represents a screening of some Romanian raw honeys collected from different geographic area and investigated the chemical characterization, microbiological examination and probiotic effect for these honeys. The general effects of geographic origin on the chemical and microbiological characterization for honey have been reported previously [1; 5; 6; 7; 8; 16; 30; 51; 52; 53; 54]. From the ten different geographical locations, the botanical origin shows similarities, and, according to palynological examination and chemical characterization, the samples can be framed as polyfloral honey. Generally, this type of honey has water content below 20% [51; 52], which can be observed also in this study. Regarding the sugar spectrum, the higher concentrations in fructose and glucose was registered for west (P9) and south-east (P8) of the country, while sucrose was in higher amounts in north-west of the country (P5 and P1). Fructooligosaccharides (sucrose), but also other oligosaccharides, can contribute as a nutritional ingredient, together with other prebiotic bacteria [9] from honeybee gut, to growing the probiotic bacteria, which have efficacy against pathogens [55]. Another important parameter for stimulating the growth and/or activity of the probiotic bacteria is the acidity. These bacteria need low acidity for development. The raw honey samples with low acidity and pH was registered in north-west (P1) and south-east (P8) of the country. The free acidity content of honey describes the presence and amount of organic acids, like butyric, acetic, formic, lactic, succinic, pyrogtumamic, malic, and citric acids in equilibrium with their corresponding lactones, or internal esters, and some inorganic ions, such as phosphate [56; 57]. Higher values were registered in samples from south-east (P8) of the country. The quality of honey is very important when this bee product is used as  
Figure 7: Antimicrobial activity of Bacillus spp. strains (1 = B. subtilis, 2 = B. thuringiensis 1, 3 = B. mycooides, 4 = B. amyloliquefaciens, 5 = B. velezensis 1, 6 = B. velezensis 2, 7 = B. thuringiensis 2, 8 = B. velezensis 3, N = negative control-central) against Enterococcus faecalis (**a**), Staphylococcus aureus (**b**), Listeria monocytogenes (**c**), Escherichia coli (**d**), Salmonella enteritidis (**e**), Pseudomonas aeruginosa (**f**). natural antibiotic or as a nutritional supplement. In this context, we evaluated the HMF, diastase, contaminants from the class of antibiotics, and heavy metals from analyzed honey samples. In addition, in the north-west (P3) and in the center (P10) of the country, we found higher levels of Zn. This microelement together with the prebiotic and probiotic bacteria from honey increasing its antibacterial activity. After the microbiological examination seven _Bacillus spp._ isolates were identified in the raw honey samples: in the north-west (P1, P5) of the country: _Bacillus mycoides_ and _Bacillus amyloliquefaciens_; in the north (P4) of the country: _Bacillus thuringiensis_; in the west of the country (P9): _Bacillus subtilis_, _Bacillus velezensis_; and in the center (P10) of the country: _B. thuringiensis_, _Bacillus velezensis_. _Leuconostoc mesenteroides_ ssp _cremoris_[58] was identified in a single sample. It is recognized as a potential probiotic, and its effect on _Candida albicans_[59] was studied before. Probiotic bacteria can be found in raw, unfiltered honey, including Lactobacillus and Bifidobacteria [17]. [PERSON] et al. (2012) [17] also found and isolated from different honey sources, probiotic bacteria, such as _B.subtilis_, _B.licheniforms_, _B. amyloliquefaciens_, _B. thuringiensis_, _B. cereus_, _B. pseudomycoides_, and _B. mycoides_. In the present study, no Lactobacillus or Bifidobacteria were identified. In addition, opportunistic pathogens were identified: _Ewingella americana_[60], _Staphylococcus hominis_ ssp. _Hominis_[61], _S. haemolyticus_, _S. uarnieri_, _S.