Export file:


  • RIS(for EndNote,Reference Manager,ProCite)
  • BibTex
  • Text


  • Citation Only
  • Citation and Abstract

Biofilm formation and adhesion to bovine udder epithelium of potentially probiotic lactic acid bacteria

Faculty II, Department for Bioprocess Engineering and Microbiology of the University of Applied Sciences and Arts Hannover, Lower Saxony, Germany

Special Issues: Bacterial adherence: much more than a bond

Mastitis is one of the most important diseases threatening modern dairy herds. The idea of fighting the disease through colonising the udder with lactic acid bacteria (LAB), thereby building a beneficial biofilm, is the base for a probiotic approach towards mastitis control. The purpose of this study was to screen 13 LAB strains (eleven wild strains, two ATCC strains) inhibitory to the growth of mastitis-causing pathogens for their in vitro ability to form a biofilm and to adhere to bovine glandular mammary epithelium in order to assess their probiotic potential. Furthermore, we aimed to gain knowledge about the chemical nature of the adhesins involved by subjecting the bacteria to various chemical and enzymatical pre-treatments. The biofilms were grown on hydrophilic glass and on hydrophobic polypropylene in de Man, Rogosa and Sharpe (MRS) broth and afterwards quantified with a crystal violet assay. Biofilm formation was observed in all strains. However, the extent strongly depended on the strain, surface charge and medium. The adhesion assay also revealed a strong strain dependency, but this trait was also present in all of the investigated LAB isolates. Depending on the strain, chemical or enzymatical pre-treatment revealed carbohydrate molecules as well as proteins and lipids to be crucial for the adhesion of LAB to epithelial cells. The seven strains showing the strongest biofilm formation and/or adhesion represent promising candidates for further investigation in order to develop a probiotic remedy for the treatment of mastitis. Still, their safety for consumers and patients as well as their capability to colonise the udder remain to be investigated in in vivo studies.
  Article Metrics

Keywords probiotic potential; biofilm; adhesion; lactic acid bacteria; mastitis

Citation: Jonathan K. Wallis, Volker Krömker, Jan-Hendrik Paduch. Biofilm formation and adhesion to bovine udder epithelium of potentially probiotic lactic acid bacteria. AIMS Microbiology, 2018, 4(2): 209-224. doi: 10.3934/microbiol.2018.2.209


  • 1. De VS, Fox LK, Piepers S, et al. (2012) Invited review: Mastitis in dairy heifers: Nature of the disease, potential impact, prevention, and control. J Dairy Sci 95: 1025–1040.    
  • 2. Rollin E, Dhuyvetter KC, Overton MW (2015) The cost of clinical mastitis in the first 30 days of lactation: An economic modeling tool. Prev Vet Med 122: 257–264.    
  • 3. Schönborn S, Krömker V (2016) Detection of the biofilm component polysaccharide intercellular adhesin in Staphylococcus aureus infected cow udders. Vet Microbiol 196: 126–128.    
  • 4. Varhimo E, Varmanen P, Fallarero A, et al. (2011) Alpha- and beta-casein components of host milk induce biofilm formation in the mastitis bacterium Streptococcus uberis. Vet Microbiol 149: 381–389.    
  • 5. Donlan RM (2002) Biofilms: Microbial life on surfaces. Emerg Infect Dis 8: 881–890.    
  • 6. An YH, Friedman RJ (1998) Concise review of mechanisms of bacterial adhesion to biomaterial surfaces. J Biomed Mater Res 43: 338–348.    
  • 7. Anderl JN, Franklin MJ, Stewart PS (2000) Role of antibiotic penetration limitation in Klebsiella pneumoniae biofilm resistance to ampicillin and ciprofloxacin. Antimicrob Agents Ch 44: 1818–1824.    
  • 8. Zahller J, Stewart PS (2002) Transmission electron microscopic study of antibiotic action on Klebsiella pneumoniae biofilm. Antimicrob Agents Ch 46: 2679–2683.    
  • 9. Schönborn S, Wente N, Paduch JH, et al. (2017) In vitro ability of mastitis causing pathogens to form biofilms. J Dairy Res 84: 198–201.    
  • 10. Klostermann K, Crispie F, Flynn J, et al. (2008) Intramammary infusion of a live culture of Lactococcus lactis for treatment of bovine mastitis: Comparison with antibiotic treatment in field trials. J Dairy Res 75: 365–373.
  • 11. Espeche MC, Pellegrino M, Frola I, et al. (2012) Lactic acid bacteria from raw milk as potentially beneficial strains to prevent bovine mastitis. Anaerobe 18: 103–109.    
  • 12. Frola ID, Pellegrino MS, Espeche MC, et al. (2012) Effects of intramammary inoculation of Lactobacillus perolens CRL1724 in lactating cows' udders. J Dairy Res 79: 84–92.    
  • 13. Bajagai YS, Klieve AV, Dart V, et al. (2016) Probiotics in animal nutrition: production, impact and regulation. FAO Animal Production and Health Paper, Rome: Food and Agriculture Organization of the United Nations (FAO).
  • 14. Reid G, Sanders ME, Gaskins HR, et al. (2003) New scientific paradigms for probiotics and prebiotics. J Clin Gastroenter 37: 105–118.    
  • 15. Hagi T, Sasaki K, Aso H, et al. (2013) Adhesive properties of predominant bacteria in raw cow's milk to bovine mammary gland epithelial cells. Folia Microbiol 58: 515–522.    
  • 16. Crispie F, Alonsogómez M, O'Loughlin C, et al. (2008) Intramammary infusion of a live culture for treatment of bovine mastitis: Effect of live lactococci on the mammary immune response. J Dairy Res 75: 374–384.
  • 17. Beecher C, Daly M, Berry DP, et al. (2009) Administration of a live culture of Lactococcus lactis DPC 3147 into the bovine mammary gland stimulates the local host immune response, particularly IL-1β and IL-8 gene expression. J Dairy Res 76: 340.    
  • 18. Tallon R, Arias S, Bressollier P, et al. (2007) Strain- and matrix-dependent adhesion of Lactobacillus plantarum is mediated by proteinaceous bacterial compounds. J Appl Microbiol 102: 442–451.
  • 19. Bouchard DS, Rault L, Berkova N, et al. (2013) Inhibition of Staphylococcus aureus invasion into bovine mammary epithelial cells by contact with live Lactobacillus casei. Appl Environ Microb 79: 877–885.    
  • 20. Rzhepishevska O, Hakobyan S, Ruhal R, et al. (2013) The surface charge of anti-bacterial coatings alters motility and biofilm architecture. Biomater Sci 1: 589–602.    
  • 21. Diepers AC, Krömker V, Zinke C, et al. (2017) In vitro ability of lactic acid bacteria to inhibit mastitis-causing pathogens. Sustain Chem Pharm 5: 84–92.    
  • 22. Terraf MC, Tomás J, Nader-Macías MEF, et al. (2012) Screening of biofilm formation by beneficial vaginal lactobacilli and influence of culture media components. J Appl Microbiol 113: 1517–1529.    
  • 23. Otero MC, Nader-Macías ME (2007) Lactobacillus adhesion to epithelial cells from bovine vagina, In: Méndez-Vilas A, Editor, Communicating current research and educational topics and trends in applied microbiology, Badajoz: Formatex, 749–757.
  • 24. Greene JD, Klaenhammer TR (1994) Factors involved in adherence of lactobacilli to human Caco-2 cells. Appl Environ Microb 60: 4487–4494.
  • 25. Roos S, Jonsson H (2002) A high-molecular-mass cell-surface protein from Lactobacillus reuteri 1063 adheres to mucus components. Microbiology 148: 433–442.    
  • 26. Wasko A, Polak-Berecka M, Paduch R, et al. (2014) The effect of moonlighting proteins on the adhesion and aggregation ability of Lactobacillus helveticus. Anaerobe 30: 161–168.    
  • 27. Iturralde M, Aguilar B, Baselga R, et al. (1993) Adherence of ruminant mastitis Staphylococcus aureus strains to epithelial cells from ovine mammry gland primary cultures and from a rat intestinal cell line. Vet Microbiol 38: 115–127.    
  • 28. Zobell CE (1943) The effect of solid surfaces upon bacterial activity. J Bacteriol 46: 39–56.
  • 29. Li XJ, Yue LY, Guan XS, et al. (2008) The adhesion of putative probiotic lactobacilli to cultured epithelial cells and porcine intestinal mucus. J Appl Microbiol 104: 1082.    
  • 30. Ramírez MDF, Smid EJ, Abee T, et al. (2015) Characterisation of biofilms formed by Lactobacillus plantarum WCFS1 and food spoilage isolates. Int J Food Microbiol 207: 23–29.    
  • 31. Opdebeeck JP, Frost AJ, O'Boyle D (1988) Adhesion of Staphylococcus aureus and Escherichia coli to bovine udder epithelial cells. Vet Microbiol 16: 77–86.    
  • 32. Kinoshita H, Ohuchi S, Arakawa K, et al. (2016) Isolation of lactic acid bacteria bound to the porcine intestinal mucosa and an analysis of their moonlighting adhesins. Biosci Microbiota Food Health 35: 185–196.    
  • 33. Hamadi F, Asserne F, Elabed S, et al. (2014) Adhesion of Staphylococcus aureus on stainless steel treated with three types of milk. Food Control 38: 104–108.    
  • 34. Dat NM, Hamanaka D, Tanaka F, et al. (2010) Surface conditioning of stainless steel coupons with skim milk solutions at different pH values and its effect on bacterial adherence. Food Control 21: 1769–1773.    
  • 35. Plumed-Ferrer C, Uusikyla K, Korhonen J, et al. (2013) Characterization of Lactococcus lactis isolates from bovine mastitis. Vet Microbiol 167: 592–599.    


This article has been cited by

  • 1. Patrick Di Martino, Bacterial adherence: much more than a bond, AIMS Microbiology, 2018, 4, 3, 563, 10.3934/microbiol.2018.3.563
  • 2. Jonathan Wallis, Volker Krömker, Jan-Hendrik Paduch, Biofilm Challenge: Lactic Acid Bacteria Isolated from Bovine Udders versus Staphylococci, Foods, 2019, 8, 2, 79, 10.3390/foods8020079
  • 3. Angeliki Angelopoulou, Alicja K. Warda, Colin Hill, R. Paul Ross, Non-antibiotic microbial solutions for bovine mastitis – live biotherapeutics, bacteriophage, and phage lysins, Critical Reviews in Microbiology, 2019, 1, 10.1080/1040841X.2019.1648381
  • 4. Jan-Hendrik Paduch, Johanna Lücking, Elisabeth Mansion-de Vries, Claudia Zinke, Nicole Wente, Volker Krömker, Prevention of Intramammary Infections by Prepartum External Application of a Teat Dip Containing Lactic Acid Bacteria with Antimicrobial Properties in Dairy Heifers, Pathogens, 2020, 9, 4, 288, 10.3390/pathogens9040288
  • 5. Lisa F Stinson, Azhar S M Sindi, Ali S Cheema, Ching Tat Lai, Beverly S Mühlhäusler, Mary E Wlodek, Matthew S Payne, Donna T Geddes, The human milk microbiome: who, what, when, where, why, and how?, Nutrition Reviews, 2020, 10.1093/nutrit/nuaa029

Reader Comments

your name: *   your email: *  

© 2018 the Author(s), licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution Licese (http://creativecommons.org/licenses/by/4.0)

Download full text in PDF

Export Citation

Copyright © AIMS Press All Rights Reserved