To explore the regularity and characteristics of biofilm formation of Bifidobacterium, the biofilm-forming ability of 114 strains distributing in six species of Bifidobacterium, including B. longum, B. breve, B. animalis, B. bifidum, B. adolescentis, and B. pseudocatenulatum, was analyzed by microplate assay. The results showed that according to the absorbance, Bifidobacterium was divided into three types: non-biofilm formers (OD600 nm≤1), weak biofilm formers (1<OD600 nm<3) and strong biofilm formers (OD600 nm≥ 3). All strains of B. bifidum were strong biofilm formers, while B.longum strains were not strong biofilm formers, 57.90% of which were non-biofilm formers. The biofilm-forming ability of other Bifidobacterium was different, including all three types of strains. It was found that the biofilm structures of B.animalis and B.bifidum were mushroom-shaped and all of other Bifidobacterium were flat using scanning electron microscopy, and the biofilms of B.longum and B.bifidum contained a higher proportion of viable bacteria using confocal laser scanning microscopy. The effect of surface properties of Bifidobacterium on the biofilm formation was performed using the adhesion of bacteria to the solvent. The results showed that the biofilm-forming ability of B. pseudocatenulatum was only related to the Lewis acid properties, B. breve, B. animalis and B. adolescentis were positively correlated with the surface hydrophobicity and Lewis acid properties, but there was no correlation between surface properties and biofilm formation for B. longum. The strains of B. bifidum with strong biofilm-forming ability, which exhibited high hydrophobicity and Lewis acid properties, had no relation between its surface properties to its biofilm-forming ability. In summary, the biofilm-forming characteristics of Bifidobacterium exerted species-specific and strain-specific effects and related to the surface characteristics.
[1] SIVAN A, CORRALES L, HUBERT N, et al. Commensal Bifidobacterium promotes antitumor immunity and facilitates anti-PD-L1 efficacy[J]. Science, 2015, 350(6 264): 1 084-1 089.
[2] CHEN H, TIAN M, CHEN L, et al. Optimization of composite cryoprotectant for freeze-drying Bifidobacterium bifidum BB01 by response surface methodology[J]. Artificial Cells, Nanomedicine, and Biotechnology, 2019, 47(1): 1 559-1 569.
[3] SHOKRI Z, FAZELI M R, ARDJMAND M, et al. Factors affecting viability of Bifidobacterium bifidum during spray drying[J]. Journal of Faculty of Pharmacy,2015, 23(1): 7.
[4] VERRUCK S, DE CARVALHO M W, DE LIZ G R, et al. Survival of Bifidobacterium BB-12 microencapsulated with full-fat goat’s milk and prebiotics when exposed to simulated gastrointestinal conditions and thermal treatments[J]. Small Ruminant Research, 2017, 153: 48-56.
[5] CHEOW W S, HADINOTO K. Biofilm-like Lactobacillus rhamnosus probiotics encapsulated in alginate and carrageenan microcapsules exhibiting enhanced thermotolerance and freeze-drying resistance[J]. Biomacromolecules, 2013, 14(9): 3 214-3 222.
[6] BUCHER T, KARTVELISHVILY E, KOLODKIN-GAL I. Methodologies for studying B. subtilis biofilms as a model for characterizing small molecule biofilm inhibitors[J]. Journal of Visualized Experiments: JoVE, 2016 (116): e54612.:
[7] 林炳谕. 双歧杆菌的生物膜样化及其应用[D]. 无锡:江南大学,2017.
[8] JAMAL M, AHMAD W, ANDLEEB S, et al. Bacterial biofilm and associated infections[J]. Journal of the Chinese Medical Association: JCMA, 2018, 81(1): 7-11.
[9] TOYOFUKU M, INABA T, KIYOKAWA T, et al. Environmental factors that shape biofilm formation[J]. Bioscience, Biotechnology, and Biochemistry, 2016, 80(1): 7-12.
[10] THEWES N, LOSKILL P, JUNG P, et al. Hydrophobic interaction governs unspecific adhesion of staphylococci: a single cell force spectroscopy study[J]. Beilstein Journal of Nanotechnology, 2014, 5(1): 1 501-1 512.
[11] OLENA R, SHOGHIK H, ROHIT R, et al. The surface charge of anti-bacterial coatings alters motility and biofilm architecture[J]. Biomaterials Science, 2013, 1(6): 589-602.
[12] NGUYEN H D N, YANG Y S, YUK H G. Biofilm formation of Salmonella typhimurium on stainless steel and acrylic surfaces as affected by temperature and pH level[J]. LWT-Food Science and Technology, 2014,55(1): 383-388.
[13] OSS C J V, GOOD R J, CHAUDHURY M K. The role of van der Waals forces and hydrogen bonds in “hydrophobic interactions” between biopolymers and low energy surfaces[J]. Journal of Colloid & Interface Science, 1986, 111(2): 378-390.
[14] ABDALLAH M, BENOLIEL C, JAMA C, et al. Thermodynamic prediction of growth temperature dependence in the adhesion of Pseudomonas aeruginosa and Staphylococcus aureus to stainless steel and polycarbonate[J]. Journal of Food Protection, 2014, 77(7): 1 116-1 126.
[15] KHELISSA S O, JAMA C, ABDALLAH M, et al. Effect of incubation duration, growth temperature, and abiotic surface type on cell surface properties, adhesion and pathogenicity of biofilm-detached Staphylococcus aureus cells[J]. AMB Express, 2017,7(1): 191.
[16] KIELY L J, OLSONl N F, MORTENSEN G. The physicochemical surface characteristics of Brevibacterium linens[J]. Colloids and Surfaces. B: Biointerfaces, 1997, 9(6):297-304.
[17] STEPANOVIC S, VUKOVIC D, DAKIC I, et al. A modified microtiter-plate test for quantification of staphylococcal biofilm formation[J]. Journal of Microbiological Methods, 2000, 40(2): 175-179.
[18] LANDETA G, CURIEL J A, CARRASCOSA A V, et al. Technological and safety properties of lactic acid bacteria isolated from Spanish dry-cured sausages[J]. Meat Science, 2013, 95(2): 272-280.
[19] RODRIGUEZ-MELCON C, RIESCO-PELAEZ F, CARBALLO J, et al. Structure and viability of 24- and 72-h-old biofilms formed by four pathogenic bacteria on polystyrene and glass contact surfaces[J]. Food Microbiology, 2018, 76(5): 513-517.
[20] KOS B, ŠUŠKOVIC′ 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.
[21] BASSON A, FLEMMING L A, CHENIA H Y. Evaluation of adherence, hydrophobicity, aggregation, and biofilm development of Flavobacterium johnsoniae-like isolates[J]. Microbial Ecology, 2008, 55(1): 1-14.
[22] COLLOCA M E, AHUMADA M C, LÓPEZ M E, et al. Surface properties of Lactobacilli isolated from healthy subjects[J]. Oral Diseases, 2000, 6(4): 227-233.
[23] BUJNAKOVA D, KMET V. Functional properties of Lactobacillus strains isolated from dairy products[J]. Folia Microbiologica, 2012, 57(4): 263-267.
[24] TOLKER-NIELSEN T, MOLIN S. Spatial organization of microbial biofilm communities[J]. Microbial Ecology, 2000, 40(2): 75-84.
[25] SUTHERLAND I W. Biofilm exopolysaccharides: a strong and sticky framework[J]. Microbiology, 2001, 147(1): 3-9.
[26] RZHEPISHEVSKA O, HAKOBYAN S, RUHAL R, et al. The surface charge of anti-bacterial coatings alters motility and biofilm architecture[J]. Biomaterials Science, 2013, 1(6): 589-602.
[27] HASSAN A N, FRANK J F. Attachment of Escherichia coli O157:H7 grown in tryptic soy broth and nutrient broth to apple and lettuce surfaces as related to cell hydrophobicity, surface charge, and capsule production[J]. International Journal of Food Microbiology, 2004, 96(1): 103-109.
[28] AMBALAM P, KONDEPUDI K K, NILSSONil I, et al. Bile enhances cell surface hydrophobicity and biofilm formation of bifidobacterial[J]. Applied Biochemistry and Biotechnology, 2014, 172(4): 1 970-1 981.
[29] MAZUMDER S, FALKINHAM III J O, DIETRICH A M, et al. Role of hydrophobicity in bacterial adherence to carbon nanostructures and biofilm formation[J]. Biofouling, 2010, 26(3): 333-339.
[30] SPERANZA G, GOTTARDI G, PEDERZOLLI C, et al. Role of chemical interactions in bacterial adhesion to polymer surfaces[J]. Biomaterials, 2003,25(11): 2 029-2 037.
[31] SAMOT J, LEBRETON J, BADET C. Adherence capacities of oral Lactobacilli for potential probiotic purposes[J]. Anaerobe, 2011, 17(2): 69-72.
