植物乳杆菌FB-T9抑制变异链球菌及其生物膜形成的研究
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摘要
为筛选得到对变异链球菌及其生物膜形成具有抑制作用的益生菌,从健康及龋病病人口腔、健康人粪便以及新鲜菜叶等样品中分离出乳酸菌,从中挑选出12株对变异链球菌具有良好抑菌作用的菌株进行抑制变异链球菌生物膜形成实验,并对这些菌株的共聚和自聚能力、自身生物膜形成能力、表面疏水性、酸碱电荷等性质进行测定。通过PCA综合分析得到一株性能优良的植物乳杆菌FB-T9,并探究了FB-T9在不同时期介导对变异链球菌生物膜形成的影响。为考察FB-T9在口腔环境中的生存适应性,对该菌粘附唾液包裹的羟磷灰石的能力和溶菌酶的耐受能力进行了体外分析。结果表明植物乳杆菌FB-T9具有良好的抑制变异链球菌及其生物膜形成的能力,同时表现出较理想的自聚和共聚能力、表面疏水性质。菌体表面酸电荷较低、碱电荷较高,自身生物膜形成能力低。对唾液包裹的羟磷灰石的粘附率为32.93%,溶菌酶最高耐受质量浓度为1.2 mg/mL,说明植物乳杆菌FB-T9具有作为预防龋病的口腔益生菌的潜力。
In order to screen probiotics that had a inhibitory effect on Streptococcus mutans and its biofilm formation,lactic acid bacteria were isolated from oral samples and feces of healthy people and fresh vegetable leaves,among which 12 strains that could significantly inhibit the growth of S.mutans were selected to detect their abilities of inhibiting the biofilm formation of S. mutans. The abilities of auto-aggregation,co-aggregation,self-biofilm formation,surface hydrophobicity and surface acid-base charge of these strains were tested,and finally Lactobacillus plantarum FB-T9 was selected by PCA analysis. The inhibition of biofilm formation of S. mutans by FB-T9 at different time was further studied. To investigate the survival adaptation of FB-T9 in oral environment,the adhesive capacity to saliva-coated hydroxyapatite and lysozyme tolerance of FB-T9 were measured.L. plantarum could strongly inhibit the growth of S. mutans and its biofilm formation and showed ideal characters. The adhesive capacity of this strain was 32.93% and the maximum tolerated concentration of lysozyme was 1.2 mg/mL. The results showed that L. plantarum FB-T9 could be a potential probiotic to prevent dental caries.
In order to screen probiotics that had a inhibitory effect on Streptococcus mutans and its biofilm formation,lactic acid bacteria were isolated from oral samples and feces of healthy people and fresh vegetable leaves,among which 12 strains that could significantly inhibit the growth of S.mutans were selected to detect their abilities of inhibiting the biofilm formation of S. mutans. The abilities of auto-aggregation,co-aggregation,self-biofilm formation,surface hydrophobicity and surface acid-base charge of these strains were tested,and finally Lactobacillus plantarum FB-T9 was selected by PCA analysis. The inhibition of biofilm formation of S. mutans by FB-T9 at different time was further studied. To investigate the survival adaptation of FB-T9 in oral environment,the adhesive capacity to saliva-coated hydroxyapatite and lysozyme tolerance of FB-T9 were measured.L. plantarum could strongly inhibit the growth of S. mutans and its biofilm formation and showed ideal characters. The adhesive capacity of this strain was 32.93% and the maximum tolerated concentration of lysozyme was 1.2 mg/mL. The results showed that L. plantarum FB-T9 could be a potential probiotic to prevent dental caries.
引文
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[12] LI Jing,WANG Yao,DENG Maocheng. Screening and characterization of an excellent Lactobacillus isolated from Pikle[J]. Food Industry Science and Technology Magazine,2016,37(6):229-232.(in Chinese)
[13] KHAN R,ZAKIR M,KHANAM Z,et al. Novel compound from Trachyspermum ammi(Ajowan caraway)seeds with antibiofilm and antiadherence activities against Streptococcus mutans:a potential chemotherapeutic agent against dental caries[J]. Journal of Applied Microbiology,2010,109(6):2151-2159.
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[16]陈佩.益生乳酸菌的降糖作用及其机制研究[D].无锡：江南大学，2014.
[17] CUI Yanting,WANG Yang,LI Pinglan. Selection and ldentification of lactic acid bacteria strains for fermented meat starter cultures[J]. Journal of Food and Biological Technology,2011,30(2):239-244.(in Chinese)
[18] LYNCH D J,FOUNTAIN T L,MAZURKIEWICZ J E,et al. Glucan-binding proteins are essential for shaping Streptococcus mutans biofilm architecture[J]. FEMS Microbiology Letters,2007,268(2):158-165.
[19] ZHAO Hongping,Wu Buling,Su Linyun,et al. In vitro investigation of the adhension properties of glucan-binding protein D gene inactivated mutant of Streptococcus mutans[J]. Journal of Clinical Oral Medicine,2012,28(6):604-607.(in Chinese)
[20] WANG Qiangyi,WANG Shuai,DONG Li. Identification of vaginal Lactobaciluus isolated by 16S r DNA saquencing and characteriztion of the hydrogen peroxide production and autoaggregation activities of the predominant strain[J]. Chinese Journal of Microecology,2014,26(10):1117-1122.(in Chinese)
[21] XUE Long,WANG Lihe,LIE Hongbing,et al. Antibacterial effects of Lactobacillus salivarius W22a on the periodontal pathogens[J]. Journal of Oral Science Research,2016,32(10):1042-1046.(in Chinese)
[22] TORLAKOVIC L,KLEPACCERAJ V,OGAARD B,et al. Microbial community succession on developing lesions on human enamel[J]. Journal of Oral Microbiology,2011,4:16125-16132.
[23] RIBEIRO S C,O'CONNOR P M,ROSS R P,et al. An anti-listerial Lactococcus lactis strain isolated from Azorean Pico cheese produces lacticin 481[J]. International Dairy Journal,2016,63:18-28.
[24] FAVARO L,BASAGLIA M,CASELLA S,et al. Bacteriocinogenic potential and safety evaluation of non-starter Enterococcus faecium strains isolated from home made white brine cheese[J]. Food Microbiology,2014,38(2):228-239.
[25] TANG Zisheng,ZHU Min,LIU Zheng. The observation of the architecture of Streptococcus mutans biofilms[J]. Chinese Journal of Microbiology and Immunology,2004,24(2):103-106.(in Chinese)
[26] YANG Juan,OU Cuocheng,CHEN Jian,et al. Screening of oral lactobacilli and research of its probiotic properties[J]. Journal of Microbiology,2013,53(4):403-408.(in Chinese)
[2] MARSH P D. Dental plaque as a biofilm and a microbial community-implications for health and disease[J]. BMC Oral Health,2006,6(1):S14.
[3] WANG Chenglong,SU Donghua,LIU Jjiaojiao,et al. Isolation and identification of Streptococcus mutans strains with different genotype from clinical samples[J]. West China Journal of Stomatology,2013,31(1):80-85.(in Chinese)
[4] SHI Airong. The characteristics and situation of prevention and control of domestic children's dental decay[J]. Youjiang Medical Journal,2014,42(4):487-490.(in Chinese)
[5] QU Yanmei. The harm and prevention and control of children's dental decay[J]. General Oral Medical Electronic Magazine,2015,7(2):14-15.(in Chinese)
[6] ZOU Jing. The risk assessment of children's dental decay[J]. West China Journal of Stomatology,2014,32(1):1-4.(in Chinese)
[7] ZHAO Chen,HUA Hong,YAN Zhimin. The research progress of probiotics for the prevention and treatment of oral diseases[J].Journal of Oral Science Research,2016,32(4):4184-4120.(in Chinese)
[8] CAGLAR E,CILDIR S K,ERGENELI S,et al. Salivary mutans streptococci and lactobacilli levels after ingestion of the probiotic bacterium Lactobacillus reuteri ATCC 55730 by straws or tablets[J]. Acta Odontologica Scandinavica,2006,64(5):314-318.
[9] MEURMAN J H,STAMATOVA I. Probiotics:contributions to oral health[J]. Oral Diseases,2007,13(5):443-451.
[10] BOSCH M,NART J,AUDIVERT S,et al. Isolation and characterization of probiotic strains for improving oral health[J].Archives of Oral Biology,2012,57(5):539-549.
[11] YAO Peilin,LIU Sisi,ZHANG Ning,et al. Screening and research of a probiotic lactic acid bacteria against dental caries in vitro[J]. Food Industry Science and Technology Magazine,2015,19:347-351.(in Chinese)
[12] LI Jing,WANG Yao,DENG Maocheng. Screening and characterization of an excellent Lactobacillus isolated from Pikle[J]. Food Industry Science and Technology Magazine,2016,37(6):229-232.(in Chinese)
[13] KHAN R,ZAKIR M,KHANAM Z,et al. Novel compound from Trachyspermum ammi(Ajowan caraway)seeds with antibiofilm and antiadherence activities against Streptococcus mutans:a potential chemotherapeutic agent against dental caries[J]. Journal of Applied Microbiology,2010,109(6):2151-2159.
[14] WEN Z T,SUNTHARALIGHAM P,CVITKOVITCH D G,et al. Trigger factor in streptococcus mutans is involved in stress tolerance,competence development,and biofilm formation[J]. Infection&Immunity,2005,73(1):219-225.
[15] SAMOT J,LEBRETON J,BADET C. Adherence capacities of oral lactobacilli for potential probiotic purposes[J]. Anaerobe,2011,17(2):69-72.
[16]陈佩.益生乳酸菌的降糖作用及其机制研究[D].无锡：江南大学，2014.
[17] CUI Yanting,WANG Yang,LI Pinglan. Selection and ldentification of lactic acid bacteria strains for fermented meat starter cultures[J]. Journal of Food and Biological Technology,2011,30(2):239-244.(in Chinese)
[18] LYNCH D J,FOUNTAIN T L,MAZURKIEWICZ J E,et al. Glucan-binding proteins are essential for shaping Streptococcus mutans biofilm architecture[J]. FEMS Microbiology Letters,2007,268(2):158-165.
[19] ZHAO Hongping,Wu Buling,Su Linyun,et al. In vitro investigation of the adhension properties of glucan-binding protein D gene inactivated mutant of Streptococcus mutans[J]. Journal of Clinical Oral Medicine,2012,28(6):604-607.(in Chinese)
[20] WANG Qiangyi,WANG Shuai,DONG Li. Identification of vaginal Lactobaciluus isolated by 16S r DNA saquencing and characteriztion of the hydrogen peroxide production and autoaggregation activities of the predominant strain[J]. Chinese Journal of Microecology,2014,26(10):1117-1122.(in Chinese)
[21] XUE Long,WANG Lihe,LIE Hongbing,et al. Antibacterial effects of Lactobacillus salivarius W22a on the periodontal pathogens[J]. Journal of Oral Science Research,2016,32(10):1042-1046.(in Chinese)
[22] TORLAKOVIC L,KLEPACCERAJ V,OGAARD B,et al. Microbial community succession on developing lesions on human enamel[J]. Journal of Oral Microbiology,2011,4:16125-16132.
[23] RIBEIRO S C,O'CONNOR P M,ROSS R P,et al. An anti-listerial Lactococcus lactis strain isolated from Azorean Pico cheese produces lacticin 481[J]. International Dairy Journal,2016,63:18-28.
[24] FAVARO L,BASAGLIA M,CASELLA S,et al. Bacteriocinogenic potential and safety evaluation of non-starter Enterococcus faecium strains isolated from home made white brine cheese[J]. Food Microbiology,2014,38(2):228-239.
[25] TANG Zisheng,ZHU Min,LIU Zheng. The observation of the architecture of Streptococcus mutans biofilms[J]. Chinese Journal of Microbiology and Immunology,2004,24(2):103-106.(in Chinese)
[26] YANG Juan,OU Cuocheng,CHEN Jian,et al. Screening of oral lactobacilli and research of its probiotic properties[J]. Journal of Microbiology,2013,53(4):403-408.(in Chinese)