筛选到能够有效改善口臭并且具有良好口腔益生特性的口腔益生菌。通过抑制H2S和挥发性硫化物的产生、降低产硫化物基因的表达量、抑制生物膜形成量等指标对乳杆菌进行筛选评估。评价2株有效的乳杆菌(植物乳杆菌CCFM1214和唾液乳杆菌CCFM1215)产H2O2的能力、自聚能力以及与具核梭杆菌的共聚能力。结果表明,植物乳杆菌CCFM1214和唾液乳杆菌CCFM1215均能显著降低具核梭杆菌的挥发性硫化物(volatile sulfur compounds, VSCs)的产生量、生物膜形成量及生长,效果显著优于唾液链球菌K12,并且有良好的自聚、共聚的能力。经过口腔益生特性评价,植物乳杆菌CCFM1214和唾液乳杆菌CCFM1215均有作为改善口臭的口腔益生菌的应用潜力。
Halitosis, also known as bad breath or oral malodor, refers to the occurrence of foul-smelling gases when breathing. Halitosis is not only an oral disease, but also has negative impact on patients′ social communication and mental health. Halitosis is caused by the decomposition of sulfur-containing amino acids or serum by oral anaerobic microorganisms, resulting in metabolites dominated by volatile sulfur compounds (VSCs), especially H2S. Fusobacterium nucleatum is one of the main H2S producing strain. Probiotics can inhibit the growth and biofilm formation of pathogens, regulate oral flora, and it is reported to be good for oral health. The aim of this study was to screen oral probiotics which can effectively improve halitosis from 148 Lactobacillus strains isolated from difference sources. Among them, 28 strains had the ability to inhibit F. nucleatum producing H2S. Then the 28 Lactobacillus strains were evaluated by the effects on the VSCs producing, the Fn1220 gene expression and the biofilm formation ability of F. nucleatum. The commercial probiotic Streptococcus saliva K12 was set as a control. Two strains, Lactiplantibacillus plantarum CCFM1214 and Ligilactobacillus salivarius CCFM1215 were selected due to their significant inhibition rate of 90.94% and 90.19%, respectively. At the same time, the inhibition effect of S. saliva K12 was 71.16%. The supernatant of L. plantarum CCFM1214 and L. salivarius CCFM1215 could also inhibit the expression of Fn1220 gene and the formation of F. nucleatum biofilm, which were significantly better than S. saliva K12. As a result, L. plantarum CCFM1214 and L. salivarius CCFM1215 could both inhibit the growth of F. nucleatum. Besides, they also had good self-aggregation, co-aggregation ability and H2O2 production ability. After the evaluation of oral probiotics characteristics, they had the application potential as oral probiotics to improve halitosis. These results contribute to the application of probiotics in the field of oral cavity and the development of the diversity of probiotic strains in functional food.
[1] LU H X, CHEN X L, WONG M C M, et al.Oral health impact of halitosis in Chinese adults[J].International Journal of Dental Hygiene, 2017, 15(4):e85-e92.
[2] LIU X N, SHINADA K, CHEN X C, et al.Oral malodor-related parameters in the Chinese general population[J].Journal of Clinical Periodontology, 2006, 33(1):31-36.
[3] PERSSON S, EDLUND M B, CLAESSON R, et al.The formation of hydrogen sulfide and methyl mercaptan by oral bacteria[J].Oral Microbiology and Immunology, 1990, 5(4):195-201.
[4] BASIC A, BLOMQVIST M, DAHLÉN G, et al.The proteins of Fusobacterium spp.involved in hydrogen sulfide production from L-cysteine[J].BMC Microbiology, 2017, 17(1):61.
[5] PIANOTTI R, LACHETTE S, DILLS S.Desulfuration of cysteine and methionine by Fusobacterium nucleatum[J].Journal of Dental Research, 1986, 65(6):913-917.
[6] OOI X J, TAN K S.Reduced glutathione mediates resistance to H2S toxicity in oral Streptococci[J].Applied and Environmental Microbiology, 2016, 82(7):2 078-2 085.
[7] LODHIA P, YAEGAKI K, KHAKBAZNEJAD A, et al.Effect of green tea on volatile sulfur compounds in mouth air[J].Journal of Nutritional Science and Vitaminology, 2008, 54(1):89-94.
[8] BOSY A, KULKARNI G V, ROSENBERG M, et al.Relationship of oral malodor to periodontitis:Evidence of independence in discrete subpopulations[J].Journal of Periodontology, 1994, 65(1):37-46.
[9] HAUKIOJA A.Probiotics and oral health[J].European Journal of Dentistry, 2010, 4(3):348-355.
[10] MOUSQUER C R, DELLA BONA A, MILANI D C, et al.Are Lactobacillus salivarius G60 and inulin more efficacious to treat patients with oral halitosis and tongue coating than the probiotic alone and placebo? A randomized clinical trial[J].Journal of Periodontology, 2020, 91(6):775-783.
[11] LEE S H, BAEK D H.Effects of Streptococcus thermophilus on volatile sulfur compounds produced by Porphyromonas gingivalis[J].Archives of Oral Biology, 2014, 59(11):1 205-1 210.
[12] YOO H J, JWA S K, KIM D H, et al.Inhibitory effect of Streptococcus salivarius K12 and M18 on halitosis in vitro[J].Clinical and Experimental Dental Research, 2020, 6(2):207-214.
[13] 王小玉. 柠檬精油对口臭抑制作用的研究[D].天津:天津医科大学, 2017.
WANG X Y.Inhibitation of lemon essential oil on halitosis[D].Tianjin:Tianjin Medical University, 2017.
[14] KOS B, SUSKOVIC′ J, VUKOVIC′ S, et al.Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92[J].Journal of Applied Microbiology, 2003, 94(6):981-987.
[15] 王江. 健康妇女阴道乳杆菌多样性分析及具有益生菌特征菌株的筛选[D].重庆:重庆医科大学, 2012.
WANG J.Diversity analysis of Lactobacillus in vagina and screening of particular strains with probiotic properties in healthy women[D].Chongqing:Chongqing Medical University, 2012.
[16] SUWABE K, YOSHIDA Y, NAGANO K, et al.Identification of an L-methionine γ-lyase involved in the production of hydrogen sulfide from L-cysteine in Fusobacterium nucleatum subsp.nucleatum ATCC 25586[J].Microbiology (Reading, England), 2011, 157(Pt10):2 992-3 000.
[17] YOSHIDA Y, SUWABE K, NAGANO K, et al.Identification and enzymic analysis of a novel protein associated with production of hydrogen sulfide and L-serine from L-cysteine in Fusobacterium nucleatum subsp.Nucleatum ATCC 25586[J].Microbiology, 2011, 157(7):2 164-2 171.
[18] ABDULLAH M A, ALASQAH M, SANAA M S, et al.The relationship between volatile sulfur compounds and the severity of chronic periodontitis:A cross-sectional study[J].Journal of Pharmacy & Bioallied Sciences, 2020, 12(Suppl 1):S268-S273.
[19] BERNARDI S, KARYGIANNI L, FILIPPI A, et al.Combining culture and culture-independent methods reveals new microbial composition of halitosis patients’ tongue biofilm[J].MicrobiologyOpen, 2020, 9(2):e958
[20] 钟焱婷, 杨虹.植物乳杆菌UD01对具核梭杆菌生物膜形成的抑制[J].生物学杂志, 2021, 38(1):36-40.
ZHONG Y T, YANG H.Inhibition of Lactobacillus plantarum UD01 on biofilm formation of Fusobacterium nucleatum[J]. Journal of Biology, 2021, 38(1):36-40.
[21] FONSECA H C, DE SOUSA MELO D, RAMOS C L, et al.Probiotic properties of Lactobacilli and their ability to inhibit the adhesion of enteropathogenic bacteria to Caco-2 and HT-29 cells[J].Probiotics and Antimicrobial Proteins, 2021, 13(1):102-112.
[22] GRAF K, LAST A, GRATZ R, et al.Keeping Candida commensal:How Lactobacilli antagonize pathogenicity of Candida albicans in an in vitro gut model[J].Disease Models & Mechanisms, 2019, 12(9):dmm039719.
[23] REDANZ S, CHENG X, GIACAMAN R A, et al.Live and let die:Hydrogen peroxide production by the commensal flora and its role in maintaining a symbiotic microbiome[J].Molecular Oral Microbiology, 2018, 33(5):337-352.
[24] 章可可, 周学东, 徐欣.口腔中过氧化氢的来源及在微生态平衡中的作用[J].华西口腔医学杂志, 2017, 35(2):215-220.
ZHANG K K, ZHOU X D, XU X.The origin of hydrogen peroxide in oral cavity and its role in oral microecology balance[J]. West China Journal of Stomatology, 2017, 35(2):215-220.
