The honey production is one of the most important agriculture branches of Ukraine. Since the beginning of 2000 year, honey becomes one of the first products of agriculture production that was allowed to export from Ukraine to the EU countries. The present study aims to evaluate microbial composition of honey of higher and first grades from south of Ukraine, which differ in water, sugar content and acidity. The correlation beetween physicochemical properties, botanical origin and microbial composition of honey were investigated. Total aerobic count (TAC), total count of fungi (TFC) and yeast (TYC), number of sporeforming anaerobes (SFA), Bacillus cereus and Clostridium botulinus were determined. Plate dilution method with specific cultural mediums and incubation conditions was used for indication and identification of microorganisms as well for quantitative CFU count determination in 1g of honey. The results indicated that the value of TAC was from 1.4 ±0.1 x 102 to 3.7 ±0.5 x 102 CFU.g-1 in higher grade honey and from 2.1 ±0.3 x 102 to 4.6 ±0.4 x 102 CFU.g-1 in first grade honey. Average amounts of fungi and yest in the first grade honey were more than in higher grade honey and amounted 15.±0.9 CFU.g-1, 6.48 ±0.7 CFU.g-1 and 17.98 ±1.2 CFU.g-1, 8.04 ±0.8 CFU.g-1 respectively. Also it was found that, 25.8% and 37.1% of higher and first grade honey samples had sporeforming anaerobes. The more of samples with Bacillus cereus and Clostridium botulinum were among samples of first-grade honey samples. In addition, we have determined that, samples of buckwheat honey and honey from fodder crops of both grades were more contaminated with all microorganisms than samples of sunflower honey and honey from fruit and non-fruit trees.Results of this work has revealed the relationship between the number of different groups and species of microorganisms in higher and first grade honey from south of Ukraine and its physicochemical properties and botanical origin. Our research could help to substantiate the sources of contamination of honey and confirm the existing findings of scientists about the direct proportional dependence of the content of microorganisms in honey, depending on its moisture content, acidity and sugars.