Hostname: page-component-848d4c4894-2xdlg Total loading time: 0 Render date: 2024-06-25T20:09:39.109Z Has data issue: false hasContentIssue false

Screening of cell surface properties of potential probiotic lactobacilli isolated from human milk

Published online by Cambridge University Press:  02 July 2018

Namita Rokana
Department of Dairy Microbiology, College of Dairy Science and Technology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana – 141004, Punjab, India
Brij Pal Singh
Department of Dairy Microbiology, College of Dairy Science and Technology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana – 141004, Punjab, India
Nishchal Thakur
Department of Dairy Microbiology, College of Dairy Science and Technology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana – 141004, Punjab, India
Chetan Sharma
Department of Dairy Microbiology, College of Dairy Science and Technology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana – 141004, Punjab, India
Rohini Devidas Gulhane
Department of Dairy Microbiology, College of Dairy Science and Technology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana – 141004, Punjab, India
Harsh Panwar*
Department of Dairy Microbiology, College of Dairy Science and Technology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana – 141004, Punjab, India
*For correspondence; e-mail:


Evaluation of eleven candidate probiotic Lactobacillus strains isolated from human milk showed that some of the strains were well endowed with desirable cell surface and attachment attributes. The cell surface properties (hydrophobicity, auto-aggregation, attachment to collagen and HT-29 monolayer) of probiotic Lactobacillus species of human milk origin were compared with reference probiotic/ non-probiotic species and pathogenic strains. The bacterial adhesion to hydrocarbons (BATH) was determined using three aliphatic (Chloroform, n-Hexane and n-Octane) and two aromatic (Toluene and Xylene) solvents. Maximum affinity of Lactobacillus strains towards chloroform and toluene indicated the presence of low electron acceptor/ acidic surface components on cell surface of most of the strains. The highest value of per cent hydrophobicity was recorded with chloroform in HM1 (L. casei) (97·10 ± 3·35%) and LGG (98·92 ± 1·24%). A moderate auto-aggregation attribute was observed in all of our Lactobacillus isolates. Only HM10, HM12 and HM13 exhibited comparatively enhanced precipitation rate after 7 h of incubation period. The adhesion potential to collagen matrix was highest in LGG (26·94 ± 5·83%), followed by HM1 (11·07 ± 3·54%) and HM9 (10·85 ± 1·74%) whereas, on HT-29 cells, HM8 (14·99 ± 3·61%), HM3 (13·73 ± 1·14%) and HM1 (11·21 ± 3·18%) could adhere effectively. In this manner, we noticed that although the cell surface properties and adhesion prospective of probiotic bacteria were strain dependent, five of our isolates viz. HM1, HM3, HM8, HM9 and HM10 exhibited promising cell surface properties, which could be further targeted as indigenous probiotic.

Research Article
Copyright © Hannah Dairy Research Foundation 2018 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)


Andriantsoanirina, V, Teolis, AC, Xin, LX, Butel, MJ & Aires, J 2014 Bifidobacterium longum and Bifidobacterium breve isolates from preterm and full term neonates: comparison of cell surface properties. Anaerobe 28 212215Google Scholar
Beaussart, A, El-Kirat-Chatel, S, Herman, P, Alsteens, D, Mahillon, J, Hols, P & Dufrêne, YF 2013 Single-cell force spectroscopy of probiotic bacteria. Biophysical Journal 104 18861892Google Scholar
Beaussart, A, El-Kirat-Chatel, S, Sullan, RMA, Alsteens, D, Herman, P, Derclaye, S & Dufrêne, YF 2014 Quantifying the forces guiding microbial cell adhesion using single-cell force spectroscopy. Nature Protocols 9 10491055Google Scholar
Benoit, M, Gabriel, D, Gerisch, G & Gaub, HE 2000 Discrete interactions in cell adhesion measured by single-molecule force spectroscopy. Nature Cell Biology 2 313317Google Scholar
Bron, PA, Tomita, S, Mercenier, A & Kleerebezem, M 2013 Cell surface-associated compounds of probiotic lactobacilli sustain the strain-specificity dogma. Current Opinions in Microbiology 16 262269Google Scholar
Collado, MC, Surono, I, Meriluoto, J & Salminen, S 2007 Indigenous dadih lactic acid bacteria: cell-surface properties and interactions with pathogens. Journal of Food Science 72 M89M93Google Scholar
Damaceno, QS, Souza, JP, Nicoli, JR, Paula, RL, Assis, GB, Figueiredo, HC, Azevedo, V & Martins, FS 2017 Evaluation of potential probiotics isolated from human milk and colostrum. Probiotics and Antimicrobial Proteins 9 371379Google Scholar
Duary, RK, Rajput, YS, Batish, VK & Grover, S 2011 Assessing the adhesion of putative indigenous probiotic lactobacilli to human colonic epithelial cells. The Indian Journal of Medical Research 134 664Google Scholar
Dufrêne, YF 2015 Sticky microbes: forces in microbial cell adhesion. Trends in Microbiology 23 376382Google Scholar
García-Cayuela, T, Korany, AM, Bustos, I, de Cadiñanos, LPG, Requena, T, Peláez, C & Martínez-Cuesta, MC 2014 Adhesion abilities of dairy Lactobacillus plantarum strains showing an aggregation phenotype. Food Research International 57 4450Google Scholar
Jost, T, Lacroix, C, Braegger, CP, Rochat, F & Chassard, C 2014 Vertical mother–neonate transfer of maternal gut bacteria via breastfeeding. Environmental Microbiology 16 28912904Google Scholar
Kainulainen, V & Korhonen, TK 2014 Dancing to another tune – adhesive moonlighting proteins in bacteria. Biology 3 178204Google Scholar
Kos, B, Suskovic, J, Vukovic, S, Simpraga, M, Frece, J & Matosic, S 2003 Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92. Journal of Applied Microbiology 94 981987Google Scholar
Li, Q, Liu, X, Dong, M, Zhou, J & Wang, J 2015 Aggregation and adhesion abilities of 18 lactic acid bacteria strains isolated from traditional fermented food. International Journal of Agriculture Policy and Research 3 8492Google Scholar
Lukic, J, Strahinic, I, Milenkovic, M, Nikolic, M, Tolinacki, M, Kojic, M & Begovic, J 2014 Aggregation factor as an inhibitor of bacterial binding to gut mucosa. Microbial Ecology 68 633644Google Scholar
Mahmoudi, I, Moussa, OB, Khaldi, TEM, Kebouchi, M, Soligot, C, Le Roux, Y & Hassouna, M 2016 Functional in vitro screening of Lactobacillus strains isolated from Tunisian camel raw milk toward their selection as probiotic. Small Ruminant Research 137 9198Google Scholar
Malik, S, Petrova, MI, Claes, IJ, Verhoeven, TL, Busschaert, P, Vaneechoutte, M, Lievens, B, Lambrichts, I, Siezen, RJ, Balzarini, J & Vanderleyden, J 2013 The highly autoaggregative and adhesive phenotype of the vaginal Lactobacillus plantarum strain CMPG5300 is sortase dependent. Applied and Environmental Microbiology 79 45764585Google Scholar
Miljkovic, M, Strahinic, I, Tolinacki, M, Zivkovic, M, Kojic, S, Golic, N & Kojic, M 2015 Agglb is the largest cell-aggregation factor from Lactobacillus paracasei subsp. paracasei BGNJ1-64, functions in collagen adhesion, and pathogen exclusion in vitro. PLoS ONE 10 e0126387Google Scholar
Moles, L, Escribano, E, de Andrés, J, Montes, MT, Rodríguez, JM, Jiménez, E, Sáenz de Pipaón, M & Espinosa-Martos, I 2015 Administration of Bifidobacterium breve PS12929 and Lactobacillus salivarius PS12934, two strains isolated from human milk, to very low and extremely low birth weight preterm infants: a pilot study. Journal of Immunology Research 2015 538171 doi: 10.1155/2015/53171Google Scholar
Nishiyama, K, Nakazato, A, Ueno, S, Seto, Y, Kakuda, T, Takai, S, Yamamoto, Y & Mukai, T 2015 Cell surface-associated aggregation-promoting factor from Lactobacillus gasseri SBT2055 facilitates host colonization and competitive exclusion of Campylobacter jejuni. Molecular Microbiology 98 712726Google Scholar
Nivoliez, A, Veisseire, P, Alaterre, E, Dausset, C, Baptiste, F, Camarès, O, Paquet-Gachinat, M, Bonnet, M, Forestier, C & Bornes, S 2015 Influence of manufacturing processes on cell surface properties of probiotic strain Lactobacillus rhamnosus Lcr35®. Applied Microbiology and Biotechnology 99 399411Google Scholar
Pérez-Cano, FJ, Dong, H & Yaqoob, P 2010 In vitro immunomodulatory activity of Lactobacillus fermentum CECT5716 and Lactobacillus salivarius CECT5713: two probiotic strains isolated from human breast milk. Immunobiology 215 9961004Google Scholar
Pisano, MB, Viale, S, Conti, S, Fadda, ME, Deplano, M, Melis, MP, Deiana, M & Cosentino, S 2014 Preliminary evaluation of probiotic properties of Lactobacillus strains isolated from Sardinian dairy products. BioMed Research International 2014 286390 doi: 10.1155/2014/286390Google Scholar
Piwat, S, Sophatha, B & Teanpaisan, R 2015 An assessment of adhesion, aggregation and surface charges of Lactobacillus strains derived from the human oral cavity. Letters in Applied Microbiology 61 98105Google Scholar
Rokana, N, Mallappa, RH, Batish, VK & Grover, S 2016 Interaction between putative probiotic Lactobacillus strains of Indian gut origin and Salmonella: impact on intestinal barrier function. LWT-Food Science and Technology 84 851860Google Scholar
Rosenberg, M, Judes, H & Weiss, E 1983 Cell surface hydrophobicity of dental plaque microorganisms in situ. Infection and Immunity 42 831834Google Scholar
Salerno, MB, Logan, BE & Velegol, D 2004 Importance of molecular details in predicting bacterial adhesion to hydrophobic surfaces. Langmuir 20 1062510629Google Scholar
Schaer-Zammaretti, P & Ubbink, J 2003 Imaging of lactic acid bacteria with AFM – elasticity and adhesion maps and their relationship to biological and structural data. Ultramicroscopy 97 199208Google Scholar
Sengupta, R, Altermann, E, Anderson, RC, McNabb, WC, Moughan, PJ & Roy, NC 2013 The role of cell surface architecture of lactobacilli in host-microbe interactions in the gastrointestinal tract. Mediators of Inflammation 2013 237921 doi: 10.1155/2013/237921Google Scholar
Shahbazi, S, Nateghi, L & Aghababyan, A 2016 Effect of fatty acids on hydrophobicity of the cell membrane of Lactobacillus species. Applied Food Biotechnology 3 194200Google Scholar
Sharma, C, Singh, BP, Thakur, N, Gulati, S, Gupta, S, Mishra, SK & Panwar, H 2017a Antibacterial effects of Lactobacillus isolates of curd and human milk origin against food-borne and human pathogens. 3 Biotech 7 31Google Scholar
Sharma, C, Gulati, S, Thakur, N, Singh, BP, Gupta, S, Kaur, S, Mishra, SK, Puniya, AK, Gill, JPS & Panwar, H 2017b Antibiotic sensitivity pattern of indigenous lactobacilli isolated from curd and human milk samples. 3 Biotech 7 53Google Scholar
Terraf, L, Mendoza, LM, Tomás, J, Silva, C & Nader-Macías, MEF 2014 Phenotypic surface properties (aggregation, adhesion and biofilm formation) and presence of related genes in beneficial vaginal lactobacilli. Journal of Applied Microbiology 117 17611772Google Scholar
Tsuneda, S, Aikawa, H, Hayashi, H, Yuasa, A & Hirata, A 2003 Extracellular polymeric substances responsible for bacterial adhesion onto solid surface. FEMS Microbiology Letters 223 287292Google Scholar
Turpin, W, Humblot, C, Noordine, ML, Thomas, M & Guyot, JP 2012. Lactobacillaceae and cell adhesion: genomic and functional screening. PLoS ONE 7 e38034Google Scholar
Van der Mei, HC, Van de Belt-Gritter, B & Busscher, HJ 1995 Implications of microbial adhesion to hydrocarbons for evaluating cell surface hydrophobicity 2. Adhesion mechanisms. Colloids and Surfaces B: Biointerfaces 5 117126Google Scholar
Vanzieleghem, T, Couniot, N, Herman-Bausier, P, Flandre, D, Dufrêne, YF & Mahillon, J 2016 Role of ionic strength in staphylococcal cell aggregation. Langmuir 32 72777283Google Scholar
Wang, L, Qin, W, Yang, S, Zhai, R, Zhou, L, Sun, C, Pan, F, Ji, Q, Wang, Y, Gu, J & Feng, X 2015 The Adh adhesin domain is required for trimeric autotransporter Apa1-mediated Actinobacillus pleuropneumoniae adhesion, autoaggregation, biofilm formation and pathogenicity. Veterinary Microbiology 177 175183Google Scholar
Wilson, WW, Wade, MM, Holman, SC & Champlin, FR 2001 Status of methods for assessing bacterial cell surface charge properties based on zeta potential measurements. Journal of Microbiological Methods 43 153164Google Scholar
Wu, Q & Shah, NP 2014 Effects of elaidic acid, a predominant industrial trans fatty acid, on bacterial growth and cell surface hydrophobicity of lactobacilli. Journal of Food Science 79 M2485M2490Google Scholar
Xu, H, Jeong, HS, Lee, HY & Ahn, J 2009 Assessment of cell surface properties and adhesion potential of selected probiotic strains. Letters in Applied Microbiology 49 434442Google Scholar
Yadav, AK, Tyagi, A, Kaushik, JK, Saklani, AC, Grover, S & Batish, VK 2013 Role of surface layer collagen binding protein from indigenous Lactobacillus plantarum 91 in adhesion and its anti-adhesion potential against gut pathogen. Microbiological Research 168 639645Google Scholar