In order to obtain the lactic acid bacteria that can inhibit oral pathogens, nine strains of lactic acid bacteria were isolated and screened from cottage pickles in Daliangshan, Sichuan. Antibacterial tests of selected isolates were conducted using Oxford cup method, with Streptococcus mutans, Candida albicans, Fusobacterium nucleatum, and Porphyromonas gingivalis as indicator bacteria. Then 16S rRNA gene sequencing was performed to identify the screened lactic acid bacteria. The pH of SD26 supernatant and its antibacterial activity against C. albicans were measured every 4 hours. Further, the SD26 supernatant of different pH was co-cultured with C. albicans to determine its effect on the growth of C. albicans. The SD26 supernatant was treated with acid, base and salt to determine its antibacterial activity against C. albicans. Finally, this study determined the antimicrobial spectrum of SD26 supernatant against foodborne pathogens. The results showed that SD26 had the highest antibacterial activity against C. albicans among the nine isolates, and the diameter of inhibition zone was (26.63±0.30) mm. SD26 was identified as Lactobacillus plantarum. The pH value of the supernatant was 3.81 when cultured for 20 hours, and the diameter of the inhibition zone against C. albicans was the largest. Furthermore, the inhibitory effect of SD26 supernatant on C. albicans depended on low pH environment, and the salting-out neutral precipitation complex solution had no antibacterial activity. SD26 supernatant had the strongest inhibitory effect on Listeria monocytogenes CMCC54002 among foodborne pathogens, with the inhibitory zone diameter of (22.68±0.06) mm. The results indicated that organic acids were the main antibacterial substances against C. albicans in the supernatant of SD26, which lays a foundation for the application of lactic acid bacteria in the prevention and treatment of oral pathogens.
[1] KILIAN M,CHAPPLE IL C,HANNIG M,et al.The oral microbiome-an update for oral healthcare professionals[J].British Dental Journal,2016,221(10):657-666.
[2] SAMPAIO-MAIA B,CALDAS I M,PEREIRA M L,et al.Chapter four-the oral microbiome in health and its implication in oral and systemic diseases[J].Advances in Applied Microbiology,2016,97:171-210.
[3] 施文元, 周学东.人体口腔微生物群的相互作用[J].华西口腔医学杂志,2010,28(1):1-4.
SHI W Y,ZHOU X D.Interaction of human oral microbiota[J].West China Journal of Stomatology,2010,28(1):1-4.
[4] ABELES S R,ROBLES-SIKISAKA R,LY M,et al.Human oral viruses are personal,persistent and gender-consistent[J].The Isme Journal,2014,8(9):1 753-1 767.
[5] JORGENSEN M R,KRAGELUND C,JENSEN P O,et al.Probiotic Lactobacillus reuterihas antifungal effects on oral Candida species in vitro[J].Journal of Oral Microbiology,2017,9(1):274-282.
[6] 董雨馨, 张君超,张红星,等.1株产细菌素植物乳杆菌的筛选及其细菌素抑菌性质研究[J].食品与发酵工业,2016,42(7):74-79.
DONG Y X,ZHANG J C,ZHANG H X,et al.Screening of bacteriocin-producing Lactobacillus plantarum and study on antimicrobial characteristics of bacteriocin[J].Food and Fermentation Industries,2016,42(7):74-79.
[7] OUWEHAND A C.A review of dose-responses of probiotics in human studies[J].Beneficial microbes,2017,8(2):143-151.
[8] BERNARD B,GIUSEPPE P,GIOVANNI S,et al.Probiotics and oral health[J].Current Pharmaceutical Design,2012,18(34):5 522-5 531.
[9] AGGETT P J,CARLO A,IRENE A,et al.Nondigestible carbohydrates in the diets of infants and young children:A commentary by the espghan committee on nutrition[J].Journal of Pediatric Gastroenterology and Nutrition,2003,36(3):329-337.
[10] PHAM L,VAN S,ROLING W,et al.Effects of probiotic Lactobacillus salivarius W24 on the compositional stability of oral microbial communities[J].Archives of Oral Biology,2009,54(2):132-137.
[11] KOLL-KLAIS P,MANDAR R,LEIBUR E,et al.Oral lactobacilli in chronic periodontitis and periodontal health:Species composition and antimicrobial activity[J].Oral Microbiology and Immunology,2005,20(6):354-361.
[12] NASE L,HATAKKA K,SAVILAHTI E,et al.Effect of long-term consumption of a probiotic bacterium,Lactobacillus rhamnosus GG,in milk on dental caries and caries risk in children[J].Caries Research,2001,35(6):412-420.
[13] HATAKKA K,AHOLA A J,YLI-KNUUTTILA H,et al.Probiotics reduce the prevalence of oral Candida in the elderly—a randomized controlled trial[J].Journal of Dental Research,2007,86(2):125-130.
[14] GHOLIZADEH P,ESLAMI H,YOUSE M,et al.Role of oral microbiome on oral cancers,a review[J].Biomedicine & Pharmacotherapy,2016,84:552-558.
[15] D'AIMMO M R,MODESTO M,BIAVATI B.Antibiotic resistance of lactic acid bacteria and Bifidobacterium spp.isolated from dairy and pharmaceutical products[J].International Journal of Food Microbiology,2007,115(1):35-42.
[16] 刘慧.现代食品微生物学实验技术[M].北京:中国轻工业出版社,2011.
LIU H.Modern food microbiology experimental technology[M].Beijing:China Light Industry Press,2011.
[17] 李清, 王英,刘小莉,等.一株广谱抑菌活性乳酸菌的筛选及特性研究[J].微生物学通报,2015,42(2):332-339.
LI Q,WANG Y,LIU X L,et al.Screening and characterization of a broad-spectrum antibacterial active Lactobacillus[J].Microbiology China,2015,42(2):332-339.
[18] WASFI R,ABD El-RAHMAN O A,ZAFER M M,et al.Probiotic Lactobacillus sp.inhibit growth,biofilm formation and gene expression of caries-inducing Streptococcus mutans[J].Journal of Cellular and Molecular Medicine,2018,22(3):1 972-1 983.
[19] 李转羽, 郝艳芳,张红星,等.1株产乳酸菌素植物乳杆菌的鉴定及功能研究[J].中国食品学报,2015,15(2):207-213.
LI Z Y,HAO Y F,ZHANG H X,et al.Identification and function studying of a Lactobacillus plantarum producing lactobacillin[J].Journal of Chinese Institute of Food Science and Technology,2015,15(2):207-213.
[20] JANCZAREK M,BACHANEK T,MAZUR E,et al.The role of probiotics in prevention of oral diseases[J].Postepy Higieny I Medycyny Doswiadczalnej,2016,70:850-857.
[21] 张英春, 韩雪,单毓娟,等.益生菌抑制致病菌作用的机制研究进展[J].微生物学通报,2012,39(9):1 306-1 313.
ZHANG Y C,HAN X,DAN Y J,et al.Recent advances in mechanism on probiotic bacteria inhibition of pathogens[J].Microbiology China,2012,39(9):1 306-1 313.
[22] STACKEBRAND E,GOEBEL B M.Taxonomic note:A place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology[J].International Journal of Systematic Bacteriology,1994,44(4):846-849.
[23] SAMOT J,LEBRETON J,BADET C.Adherence capacities of oral lactobacilli for potential probiotic purposes[J].Anaerobe,2011,17(2):69-72.
[24] MARSH P D.Microbiologic aspects of dental plaque and dental caries[J].Dental Clinics of North America,1999,43(4):599-614.
[25] 郭夏蕾, 张健,杨贞耐.抑制口腔变异链球菌的乳酸菌筛选及其抑菌机理[J].食品科学,2016,37(19):117-122.
GUO X L,ZHANG J,YANG Z N.Screening of lactic acid bacteria for inhibiting oral Streptococcus mutans and preliminary study of antibacterial mechanism[J].Food Science,2016,37(19):117-122.
[26] BRUL S,COOTE P.Preservative agents in foods:Mode of action and microbial resistance mechanisms[J].International Journal of Food Microbiology,1999,50(1-2):1-17.
[27] HALM M,HORNBAEK T,ARNEBORG N,HALM M,et al.Lactic acid tolerance determined by measurement of intracellular pH of singles cells of Candida krusei and Saccharomyces cerevisiae isolated from fermented maize dough[J].International Journal of Food Microbiology,2003,94(1):97-103.
[28] DONG A R,HO T T,LO R,et al.A genetic diversity study of antifungal Lactobacillus plantarum isolates[J].Food Control,2017,72:83-89.
