对分离于健康儿童口咽部的2株对金黄色葡萄球菌等常见致病菌具有广谱拮抗作用的甲型链球菌菌株1-4和4-2的安全性进行评价, 为其作为呼吸道益生菌产品的研发和制备提供安全性依据。通过给予SPF级小鼠口服含有菌株1-4和4-2不同剂量的菌液的急性毒性实验以及耐药性的测定, 观察动物是否出现死亡, 急性毒性体征, 细菌移位生长等情况。未发现动物死亡情况, 实验组的体重变化采食量以及脏器指数均与对照组无显著差异, 未有细菌移位生长的现象发生。菌株1-4和4-2均对9种常见抗生素的检测结果为敏感, 其中菌株4-2对红霉素具有抗性。菌株1-4和4-2的安全性评价初步显示对小鼠无毒副作用, 具有良好的安全性, 可以作为呼吸道微生态制剂等产品的备用菌株。
The safety of strains 1-4 and 4-2, two potential probiotics isolated from the oropharynx of healthy children with broad-spectrum antagonism against Staphylococcus aureus and other common pathogenic bacteria, were evaluated so as to provide safety basis for the development and preparation of probiotics as respiratory tract products. The acute toxicity test and the determination of drug resistance of SPF mice with different doses of bacterial liquid containing strains 1-4 and 4-2 were conducted, and evaluation were made by observing death of the animals, signs of acute toxicity, bacterial translocation and growth, etc. Death of animals, or significant difference between the experimental group and the control group in weight change, food intake and organ index were not observed. Strains 1-4 and 4-2 were sensitive to 8 antibiotics tested and were resistant to cefotaxime and ceftriaxone, in addition strain 4-2 was also resistant to erythromycin. The safety evaluation of strains 1-4 and 4-2 showed that they were harmless to mice and had good safety, and could be used as backup strains for preparation of respiratory microecological and other products.
[1] PETTIGREW M M, LAUFER A S, GENT J F, et al.Upper respiratory tract microbial communities, acute otitis media pathogens, and antibiotic use in healthy and sick children[J].Applied and Environmental Microbiology, 2012, 78(17):6 262-6 270.
[2] SANDERS M E.Probiotics in 2015:Their scope and use[J].Journal of Clinical Gastroenterology, 2015, 49(1):2-6.
[3] EJTAHED H S, MOHTADI-NIA J, HOMAYOUNI-RAD A, et al.Effect of probiotic yogurt containing Lactobacillus acidophilus and Bifidobacterium lactis on lipid profile in individuals with type 2 diabetes mellitus[J].Journal of Dairy Science, 2011, 94(7):3 288-3 294.
[4] KALAM A M A, MANOBENDRO S, TIEJUN L, et al.Probiotic species in the modulation of gut microbiota:An overview[J].Biomed Research International, 2018, 2018(8):9 478 630.
[5] CITI S.Intestinal barriers protect against disease[J].Science, 2018, 359(6 380):1 097-1 098.
[6] CAGLAR E, KARGUL B, TANBOGA I.Bacteriotherapy and probiotics' role on oral health[J].Oral Diseases, 2010, 11(3):131-137.
[7] 张研, 蒋鸿阳, 刘殿媛, 等.益生菌应用对2型糖尿病患者肠道菌群的影响[J].中国老年保健医学, 2019(4):63-66.
ZHANG Y, JIANG, H Y, LIU D Y, et al.Effects of probiotics on gut microbiota in type 2 diabetes patients[J].Chinese Journal of Geriatric Care, 2019(4):63-66.
[8] Van BEVER H P, NAGARAJAN S, SHEK L P, et al.Option:Primary prevention of allergy:Will it soon become a reality?[J].Pediatr Allergy Immunol, 2016, 27(1):6-12.
[9] GAREAU M G, SHERMAN P M, WALKER W A.Probiotics and the gut microbiota in intestinal health and disease[J].Nat Rev Gastroenterol Hepatol, 2010, 7(9):503-514.
[10] URTON J P, WESCOMBE P A, MOORE C J, et al.Safety assessment of the oral cavity probiotic Streptococcus salivarius K12[J].Appl Environ Microbiol, 2006, 72(4):3 050-3 053.
[11] CARLSSON J, GRAHNEN H, JONSSON G, et al.Early establishment of Streptococcus salivarius in the mouth of infants[J].Journal of Dental Research, 1970, 49(2):415.
[12] KAZOR C E, MITCHELL P M, LEE A M, et al.Diversity of bacterial populations on the tongue dorsa of patients with halitosis and healthy patients[J].Journal of Clinical Microbiology, 2003, 41(2):558-563.
[13] JOHANSON W G JR, BLACKSTOCK R, PIERCE A K, et al.The role of bacterial antagonism in pneumococcal colonization of the human pharynx[J].Lab Clin Med, 1970, 75(6):946-952.
[14] BIDOSSI A, GRANDI D R, TOSCANO M, et al.Probiotics Streptococcus salivarius 24SMB and Streptococcus oralis 89a interfere with biofilm formation of pathogens of the upper respiratory tract[J].BMC Infect Dis, 2018, 18(2):653.
[15] 王冲, 何亮, 曾忠铭.小鼠口咽部甲型链球菌的微生态研究[J].中华全科医学, 2010, 8(8):944-946.
WANG C, HE L, ZENG Z M.Study on oropharyngeal ecosystem of alpha-haemolytic Streptococci[J].Chinese Journal of General Practice, 2010, 8(8):944-946.
[16] ROOS K, HAKANSSON E G, HOLM S.Effect of recolonisation with ‘interfering' α Streptococci on recurrences of acute and secretory otitis media in children:Randomised placebo controlled trial[J].British Medical Journal, 2001, 322(7 280):210-212.
[17] SANTAGATI M, SCILLATO M, PATANÈ F, et al.Bacteriocin-producing oral Streptococci and inhibition of respiratory pathogens[J].Pathogens & Disease, 2012, 65(1):23-31.
[18] 王坚镪, 丁在咸, 张旻.益生菌对炎症性肠病小鼠肠道菌群紊乱及细菌移位的影响[J].上海交通大学学报(医学版), 2010(2):186-190.
WANG J Q, DING Z X, ZHANG M.Effect of probiotics on intestinal flora disturbance and bacterial translocation in mice with spontaneous colitis[J].Journal of Shanghai Jiaotong University, 2010(2):186-190.
[19] 冯大伟, 周家春.益生乳酸菌的纸片扩散法药敏性试验评价[J].微生物学通报, 2010, 37(3):454-464.
FENG D W, ZHOU J C.Evaluation of agar disc diffusion method for determining antibiotic susceptobility of lactic acid bacteria strains by a selected culture medium[J].Microbiology China, 2020, 37(3):454-464.
