Adhesion Capacity of Weissella cibaria to Bovine Mammary Tissue and the Effect of Bio-Sealant Topical Application on Physicochemical Properties of Milk


The ability of a probiotic strain (Weissella cibaria) to adhere on tissue and the effect of its topical application in nipples of lactating cows on physicochemical characteristics of milk were evaluated. An ex vivo model was used to demonstrate the adhesion capacity of W. cibaria. Tissue samples were randomly distributed in three different solutions corresponding to three treatments (a nipple bio-sealant formulation, sterile PBS solution and biomass of W. cibaria, sterile PBS solution without microorganism addition). The number of bacteria adhered in tissue was quantified and observed using electron microscopy. Additionally, a bio-sealant prepared with W. cibaria was topically applied to nipples of dairy cows. Milk samples were taken every 7 days for 60 days. Two controls were used. California mastitis test (CMT), somatic cell count, electrical conductivity, pH, density, and acidity were evaluated. The adhesion capacity of W. cibaria strain to epithelial cells of bovine teat tissue samples was demonstrated. When the strain was added as a bio-sealant, the adhesion capacity of W. cibaria was 80.44%. The response variables did not show significant differences among treatments; these results indicate the safety of the topical application of W. cibaria on the bovine mammary gland. In this study, a new safe way of administering probiotic microorganisms in nipples of lactating cows was demonstrated. W. cibaria adheres to the bovine mammary tissue and can be topically applied in nipples of lactating cows without affecting the physicochemical characteristics of milk.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2


  1. 1.

    Ren D, Li C, Qin Y, Yin R, Li X, Tian M, et al (2012) Inhibition of Staphylococcus aureus adherence to Caco-2 cells by lactobacilli and cell surface properties that influence attachment. Anaerobe [Internet]. [cited 2017 Jan 11] 18(5):508–15. Available from:

  2. 2.

    Pringsulaka O, Rueangyotchanthana K, Suwannasai N, Watanapokasin R, Amnueysit P, Sunthornthummas S, et al (2015) In vitro screening of lactic acid bacteria for multi-strain probiotics. Livest Sci [Internet]. Elsevier; [cited 2016 May 9];174:66–73. Available from:

  3. 3.

    García-Ruiz A, González de Llano D, Esteban-Fernández A, Requena T, Bartolomé B, Moreno-Arribas MV (2014) Assessment of probiotic properties in lactic acid bacteria isolated from wine. Food Microbiol [Internet]. [cited 2017 Jan 11];44:220–5. Available from:

  4. 4.

    Blajman J, Gaziano C, Zbrun MV, Soto L, Astesana D, Berisvil A, et al (2015) In vitro and in vivo screening of native lactic acid bacteria toward their selection as a probiotic in broiler chickens. Res Vet Sci [Internet]. Elsevier Ltd; [cited 2017 Jan 12];101:50–6. Available from:

  5. 5.

    Hynönen U, Kant R, Lähteinen T, Pietilä TE, Beganovi J, Smidt H et al (2014) Functional characterization of probiotic surface layer protein-carrying Lactobacillus amylovorus strains. BMC Microbiol 14:2–16

    Article  Google Scholar 

  6. 6.

    Govender M, Choonara YE, Kumar P, Toit LC, Van Vuuren S, Pillay V (2014) Review article: A review of the advancements in probiotic delivery: conventional vs. non-conventional formulations for intestinal flora supplementation. AAPS PharmSciTech 15(1):29–43

  7. 7.

    Sheil B, McCarthy J, O’Mahony L, Bennett MW, Ryan P, Fitzgibbon JJ, et al (2004) Is the mucosal route of administration essential for probiotic function? Subcutaneous administration is associated with attenuation of murine colitis and arthritis. Gut [Internet]. [cited 2017 Jan 16] 53(5):694–700. Available from:

  8. 8.

    Serna-cock L, Enríquez CE, Campos Gaona R, Vásquez A (2013) Response of the bovine mammary gland after Weissella confusa infusion to control Streptococcus agalactiae. Rev Colomb Cienc Pecu 26:280–287

  9. 9.

    Sánchez-González L, Quintero Saavedra JI, Chiralt A (2014) Antilisterial and physical properties of biopolymer films containing lactic acid bacteria. Food Control [Internet]. Elsevier Ltd; [cited 2017 Jan 12] 35(1):200–6. Available from:

  10. 10.

    Serna-Cock L, Pabón-Rodríguez OV (2016) Development of a teat bio-sealant and evaluation of its technological and functional properties. Probiotics & Antimicro. Prot [Internet]. [cited 2016 Jun 23] 8(2):111–9. Available from:

  11. 11.

    Serna-cock L, Valencia LJ, Campos-Gaona R (2011) Bacterias acido lacticas con actividad antimicrobiana contra patogenos causantes de mastitis bovina. Rev. Bio. Agro 9(1):97–104

  12. 12.

    Espeche MC, Otero MC, Sesma F, Nader-Macias MEF (2009) Screening of surface properties and antagonistic substances production by lactic acid bacteria isolated from the mammary gland of healthy and mastitic cows. Vet Microbiol [Internet]. [cited 2016 Jun 22] 135(3–4):346–57. Available from:

  13. 13.

    Espeche MC, Pellegrino M, Frola I, Larriestra A, Bogni C (2012) Nader-Macías MEF. Lactic acid bacteria from raw milk as potentially beneficial strains to prevent bovine mastitis. Anaerobe [Internet]. Elsevier Ltd; [cited 2017 Jan 10] 18(1):103–9. Available from:

  14. 14.

    Zupancic K, Kricksic V, Kovacevic I, Kovasevic D (2017) Influence of oral probiotic Streptococcus salivarius K12 on ear and oral cavity health in humans: systematic review. Probiotics & Antimicro. Prot 9(2):102–110

  15. 15.

    Wescombe PA, Hale JD, Heng NC, Tagg JR (2012) Developing oral probiotics from Streptococcus salivarius. Future Microbiol 7(12):1355–1371

  16. 16.

    Serna-Cock LS, Johana L, Hernández V, Campos R (2010) Cinética de fermentación y acción antimicrobiana de Weissella confusa contra Staphylococcus aureus y Streptococcus agalactiae kinetic of fermentation and antimicrobial activity of Weissella confusa against Staphylococcus aureus and Streptococcus agalactia. Revista Facultad de Ingeniería Universidad de Antioquia 55:55–65

  17. 17.

    Valencia-Hernández LJ, López-López K, Serna-Cock L (2016) Weissella cibaria fungistatic activity against Fusarium spp. affecting yellow pitahaya. American Journal of Applied Sciences 13(12):1354–1364

  18. 18.

    Ren D, Li C, Qin Y, Yin R, Li X, Tian M, et al (2012) Inhibition of Staphylococcus aureus adherence to Caco-2 cells by lactobacilli and cell surface properties that influence attachment. Anaerobe [Internet]. [cited 2016 Jun 23] 18(5):508–15. Available from:

  19. 19.

    Østerås O, Whist AC, Sølverød L (2008) The influence of iodine teat dipping and an external teat sealant in heifers on bacterial isolation from quarter milk culture obtained post-calving. Livest Sci [Internet]. Elsevier B.V.; [cited 2017 Jan 12];119(1–3):129–36. Available from:

  20. 20.

    Bhutto AL, Murray RD, Woldehiwet Z (2012) California mastitis test scores as indicators of subclinical intra-mammary infections at the end of lactation in dairy cows. Res Vet Sci [Internet]. Elsevier Ltd; [cited 2017 Jan 12];92(1):13–7. Available from:

  21. 21.

    ICONTEC (2002) Norma Tecnica Colombiana 506. Productos Lacteos, Leche Pasteurizada. ICONTEC, Colombia, p 11

  22. 22.

    ICONTEC (2001) Norma Tecnica Colombiana 4978. Leche y Productos Lacteos, Determinación de la Acidez Titulable - Metodo de Referencia. ICONTEC, Colombia, p 9

  23. 23.

    Ayeni FA, Sánchez B, Adeniyi BA, de Los Reyes-Gavilán CG, Margolles A, Ruas-Madiedo P (2011) Evaluation of the functional potential of Weissella and Lactobacillus isolates obtained from Nigerian traditional fermented foods and cow’s intestine. Int J Food Microbiol [Internet]. Elsevier B.V.; [cited 2017 Jan 12];147(2):97–104. Available from:

  24. 24.

    Diepers A-C, Krömker V, Zinke C, Wente N, Pan L, Paulsen K, et al (2016) In vitro ability of lactic acid bacteria to inhibit mastitis-causing pathogens. Sustainable Chemistry and Pharmacy [Internet]. Elsevier; [cited 2017 Jan 12];0–1. Available from:

  25. 25.

    Jensen H, Grimmer S, Naterstad K, Axelsson L (2012) In vitro testing of commercial and potential probiotic lactic acid bacteria. Int J Food Microbiol [Internet]. Elsevier B.V.; [cited 2017 Jan 12];153(1–2):216–22. Available from:

  26. 26.

    Nam H, Ha M, Bae O, Lee Y (2002) Effect of Weissella confusa strain PL9001 on the adherence and growth of helicobacter pylori. Appl Environ Microbiol 68(9):4642–4645

    CAS  Article  Google Scholar 

  27. 27.

    Lee Y (2005) Characterization of Weissella kimchii PL9023 as a potential probiotic for women. FEMS Microbiol Lett [Internet]. [cited 2017 Jan 12];250(1):157–62. Available from:

  28. 28.

    Ruas-Madiedo P, Gueimonde M, Margolles A, de los Reyes-Gavilán CG, Salminen S (2006) Exopolysaccharides produced by probiotic strains modify the adhesion of probiotics and enteropathogens to human intestinal mucus. J Food Prot [Internet]. 69(8):2011–5. Available from:

  29. 29.

    Zhang Y-C, Zhang L-W, Ma W, Yi H-X, Yang X, Du M, et al (2012) Screening of probiotic lactobacilli for inhibition of Shigella sonnei and the macromolecules involved in inhibition. Anaerobe [Internet]. Elsevier Ltd; [cited 2017 Jan 13] 18(5):498–503. Available from:

  30. 30.

    García-Cayuela T, Korany AM, Bustos I, Gómez de Cadiñanos LP, Requena T, Peláez C, et al (2014) Adhesion abilities of dairy Lactobacillus plantarum strains showing an aggregation phenotype. Food Res Int [Internet]. Elsevier Ltd; [cited 2017 Jan 13];57:44–50. Available from:

  31. 31.

    Beecher C, Daly M, Berry DP, Klostermann K, Flynn J, Meaney W, 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-1beta and IL-8 gene expression. J Dairy Res [Internet]. [cited 2017 Jan 13];76(3):340–348. Available from:

  32. 32.

    Patoo R, Singh DV, Rukshan, Kaushl S, Singh MK (2014) Compositional changes in colostrum and milk of hill cows of Uttarakhand during different lactation stages. Indian Journal of Hill Farming 27(2):54–58

    Google Scholar 

  33. 33.

    Le Maréchal C, Thiéry R, Vautor E, Le Loir Y (2011) Mastitis impact on technological properties of milk and quality of milk products—a review. Dairy Sci Technol [Internet]. [cited 2017 Jan 13];91(3):247–82. Available from:

  34. 34.

    van den Borne BHP, Vernooij JCM, Lupindu AM, van Schaik G, Frankena K, Lam TJGM, et al (2011) Relationship between somatic cell count status and subsequent clinical mastitis in Dutch dairy cows. Preventive Veterinary Medicine [Internet]. Elsevier B.V.; [cited 2017 Jan 13];102(4):265–73. Available from:

  35. 35.

    Frola ID, Pellegrino MS, Espeche MC, Giraudo JA, Nader-Macias ME, Bogni CI (2012) Effects of intramammary inoculation of Lactobacillus perolens CRL1724 in lactating cows’ udders. Journal of Dairy Research [Internet]. [cited 2017 Jan 13];79(1):84–92. Available from:

  36. 36.

    Gaggìa F, Mattarelli P, Biavati B (2010) Probiotics and prebiotics in animal feeding for safe food production. Int J Food Microbiol [Internet]. Elsevier B.V.; [cited 2016 Oct 11];141 Suppl:S15–28. Available from:

Download references


We thank the Universidad Nacional de Colombia for the financing and logistical support of this research.

Author information



Corresponding author

Correspondence to Liliana Serna-Cock.

Ethics declarations

All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

Conflict of Interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Serna-Cock, L., Pabón-Rodríguez, O.V. & Giraldo-Gómez, G.I. Adhesion Capacity of Weissella cibaria to Bovine Mammary Tissue and the Effect of Bio-Sealant Topical Application on Physicochemical Properties of Milk. Probiotics & Antimicro. Prot. 11, 1293–1299 (2019).

Download citation


  • Adhesion ability
  • Mammary tissue
  • Mastitis
  • Probiotic strain properties