Streptococcus pyogenes is a common gram-positive virulent bacterium (1). Because S. pyogenes contains various virulent factors such as streptolysin O, streptolysin S, NADase, and SpeB protease, it causes various types of infectious diseases such as pharyngitis, tonsillitis, nephritis, cellulitis, and necrotizing fasciitis (1). Because the antibiotic resistance rate of S. pyogenes has been gradually increasing worldwide (2,3), identification of novel anti-S. pyogenes therapies is of great importance. Lonicera caerulea var. emphyllocalyx (LCE) belongs to the honeysuckle family (Caprifoliaceae) and the honeysuckle genus, which is best-known for having edible berries (4). LCE is distributed in northern temperate zones such as the northern part of Japan (Hokkaido) and the northern part of the Eurasian continent (4). LCE is cultivated commercially in Japan, Canada, and Russia (4). LCE berries contain several beneficial phytochemicals, including carbohydrates, lipids, proteins, sugar, organic acids, vitamins, and mineral such as iron, magnesium, phosphorus, calcium, and potassium (5). They also Summary Streptococcus pyogenes causes several infectious diseases such as tonsillitis, cellulitis, and streptococcal toxic shock syndrome. As antibiotics are used for the general treatment of S. pyogenes infection, cases of treatment failure due to drug-resistant bacteria have increased. Lonicera caerulea var. emphyllocalyx (LCE) has been used as a folk medicine in northern Japan (Hokkaido). In this study, we investigated the antibacterial effect of methanol extracts of the fruit, stem, and leaf of LCE (LCEEs) against S. pyogenes using disk diffusion assay. As LCEE (fruit) had the strongest antibacterial activity among the three LCEEs, we focused on functional analysis of antibacterial effects of LCEE (fruit). LCEE (fruit) suppressed the growth of S. pyogenes in a dose-dependent manner. Morphological analysis by transmission electron microscopy demonstrated that LCEE (fruit) damaged the shape of S. pyogenes. Microplate and confocal laser microscopy analysis showed that biofilm formation was also suppressed by LCEE (fruit) in a dose-dependent manner. To further evaluate the surface structure of these biofilms, we performed hydrophobic analysis, which demonstrated that LCEE (fruit) reduced the hydrophobicity of the bacterial surface structure. Our data demonstrated that LCEE (fruit) had anti-bacterial and anti-biofilm effects on S. pyogenes in vitro, suggesting that the direct anti-bacterial effects of the LCEE (fruit) may be useful for treatment of local S. pyogenes infection.