Development of New Strategy for Non-Antibiotic Therapy: Bovine Lactoferrin Has a Potent Antimicrobial and Immunomodulator Effects


The antibiotic therapy has many problems, such as antibiotics resistance, hypersensitivity, direct toxicity, antibiotic-induced immunosuppresion and super-infections. This is highlighting the need for a new strategy for non-antibiotic therapy through the use of novel immunomodulators as naturally released ones (Lactoferrin). The present study investigates the potential of bovine lactoferrin (bLf), isolated from bovine milk whey, to prevent Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), Streptococcus agalactiae (S. agalactiae) and Pseudomonas aeruginosa (P. aerogenosa) growth and then evaluates its immunomodulator effect. First, bLf isolation was attempted from bovine milk whey using a cation exchange chromatography by SP-Sepharose. Second, the antimicrobial activity assays were trailed to study the antimicrobial activity of bLf. Finally, the immune effect of bLf was studied by lymphocyte transformation test. It was found that bLf was separated around molecular weight of 80 kDa and showed significant inhibitory effect against E. coli followed by P. aeruginosa, S. agalactiae and S. aureus. bLf increased lymphocyte transformations mean values in a dose dependant manner. The highest transformations mean value was determined at 50 μg/mL. In conclusion, these results suggest that bLf is a potent natural antimicrobials and immunomodulator agent.

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S. Hafez, A. Ismael, M. Mahmoud and A. Elaraby, "Development of New Strategy for Non-Antibiotic Therapy: Bovine Lactoferrin Has a Potent Antimicrobial and Immunomodulator Effects," Advances in Infectious Diseases, Vol. 3 No. 3, 2013, pp. 185-192. doi: 10.4236/aid.2013.33027.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] I. Phillips, M. Casewell, T. Cox, B. De Groot, C. Friis, R. Jones, C. Nightingale, R. Preston and J. Waddell, “Does the Use of Antibiotics in Food Animals Pose a Risk to Human Health? A Critical Review of Published Data,” Journal of Antimicrobial Chemotherapy, Vol. 53, No. 1, 2004, pp. 28-52. doi:10.1093/jac/dkg483
[2] P. J. Rajala-Schultz, K. L. Smith, J. S. Hogan and B. C. Love, “Antimicrobial Susceptibility of Mastitis Pathogens from First Lactation and Older Cows,” Veterinary Microbiology, Vol. 102, No. 1-2, 2004, pp. 33-42. doi:10.1016/j.vetmic.2004.04.010
[3] A. A. Adesiyun, S. Stoute and B. David, “Pre-Processed Bovine Milk Quality in Trinidad: Prevalence and Characteristics of Bacterial Pathogens and Occurrence of Antimicrobial Residues in Milk from Collection Centres,” Food Control, Vol. 18, No. 4, 2007, pp. 312-320. doi:10.1016/j.foodcont.2005.10.012
[4] Y. A. Suzuki, K. Shi and B. Lonnerdal, “Molecular Cloning and Functional Expression of a Human Intestinal Lactoferrin Receptor,” Biochemistry, Vol. 40, No. 51, 2001, pp. 15771-15779. doi:10.1021/bi0155899
[5] P. P. Ward, E. Paz and O. M. Conneely, “Multifunctional Roles of Lactoferrin: A Critical Overview,” Cellular and Molecular Life Sciences, Vol. 62, No. 22, 2005, pp. 2540-2548. doi:10.1007/s00018-005-5369-8
[6] W. P. Pyong, B. P. Gerald, T. H. Michael and M. Bernfield, “Exploitation of Syndecan-1 Shedding by Pseudomonas aeruginosa Enhances Virulence,” Nature, Vol. 411, 2001, pp. 98-102. doi:10.1038/35075100
[7] R. K. Chand, P. K. Sahoo, J. Kumari, B. R. Pillai and B. K. Mishra, “Dietary Administration of Bovine Lactoferrin Influences the Immune Ability of the Giant Freshwater Prawn Macrobrachium rosenbergii (de Man) and Its Resistance against Aeromonas hydrophila Infection and Nitrite Stress,” Fish & Shellfish Immunology, Vol. 21, No. 2, 2006, pp. 119-129. doi:10.1016/j.fsi.2005.10.010
[8] E. D. Robblee, P. S. Erickson, N. L. Whitehouse, A. M. McLaughlin, C. G. Schwab, J. J. Rejman and R. E. Rompala, “Supplemental Lactoferrin Improves Health and Growth of Holstein Calves during the Preweaning Phase,” Journal of Dairy Science, Vol. 86, No. 4, 2003, pp. 1458-1464. doi:10.3168/jds.S0022-0302(03)73729-1
[9] D. G. Burrin, H. Wang, J. Heath and M. A. Dudley, “Orally Administered Lactoferrin Increases Hepatic Protein Synthesis in Formula-Fed Newborn Pigs,” Pediatric Research, Vol. 40, 1996, pp. 72-76. doi:10.1203/00006450-199607000-00013
[10] K. Hayashid, T. Kaneko, T. Takeuchi, H. Shimizu, K. Ando and E. Harad, “Oral Administration of Lactoferrin Inhibits Inflammation and Nociception in Rat Adjuvant-Induced Arthritis,” Journal of Veterinary Medical Science, Vol. 66, No. 2, 2004, pp. 149-154. doi:10.1292/jvms.66.149
[11] J. Kumari and P. K. Sahoo, “Dietary Immunostimulants Influence Specific Immune Response and Resistance of Healthy and Immunocompromised Asian Catfish Clarias batrachus to Aeromonas hydrophila Infection,” Diseases of Aquatic Organism, Vol. 70, No. 1-2, 2006, pp. 63-70. doi:10.3354/dao070063
[12] K. Ishii, N. Takamura, M. Shinohara, N. Wakui, H. Shin, Y. Sumino, Y, Ohmoto, S, Teraguchi and K. Yamauchi, “Long-Term Follow-Up of Chronic Hepatitis C Patients Treated with Oral Lactoferrin for 12 Months,” Hepatology Research, Vol. 25, No. 3, 2003, pp. 226-233. doi:10.1016/S1386-6346(02)00279-6
[13] H. Tsuda, K. Sekine, K. Fujita and M. Iigo, “Cancer Prevention by Bovine Lactoferrin and Underlying Mechanisms—A Review of Experimental and Clinical Studies,” Biochemistry and Cell Biology, Vol. 80, No. 1, 2002, pp. 131-136. doi:10.1139/o01-239
[14] A. Varadhachary, J. S. Wolf, K. Petrak, B. W. O’Malley Jr., M. Spadaro, C. Curcio, G. Forni and F. Pericle, “Oral Lactoferrin Inhibits Growth of Established Tumors and Potentiates Conventional Chemo-Therapy,” International Journal of Cancer, Vol. 111, No. 3, 2004, pp. 398-403. doi:10.1002/ijc.20271
[15] M. Tomita, H. Wakabayashi, K. Shin, K. Yamau-chi, T. Yaeshima and K. Iwatsuki, “Twenty-Five Years of Research on Bovine Lactoferrin Applications,” Biochimie, Vol. 91, No. 1, 2009, pp. 52-57. doi:10.1016/j.biochi.2008.05.021
[16] H. Wakabayashi, M. Kurokawa, K. Shin, S. Teraguchi, Y. Tamura and K. Shiraki, “Oral Lactoferrin Prevents Body Weight Loss and Increases Cytokine Responses during Herpes Simplex Virus Type 1 Infection of Mice,” Bioscience, Biotechnology, and Biochemistry, Vol. 68, No. 3, 2004, pp. 537-544. doi:10.1271/bbb.68.537
[17] N. Takakura, H. Wakabayashi, H. Ishibashi, S. Teraguchi, Y. Tamura, H. Yamaguchi and S. Abe, “Oral Lactoferrin Treatment of Experimental Oral Candidiasis in Mice,” Antimicrobial Agents and Chemotherapy, Vol. 47, No. 8, 2003, pp. 2619-2623. doi:10.1128/AAC.47.8.2619-2623.2003
[18] K. Shin, H. Wakabayashi, K. Yamauchi, S. Teraguchi, Y. Tamura, M. Kurokaw and K. Shiraki, “Effects of Orally Administered Bovine Lactoferrin and Lactoperoxidase on Influenza Virus Infection in Mice,” Journal of Medical Microbiology, Vol. 54, No. 8, 2005, pp. 717-723. doi:10.1099/jmm.0.46018-0
[19] H. Wakabayashi, K. Uchida, K. Yamauchi, S. Teraguchi, H. Hayasawa and H. Yamaguchi, “Lactoferrin Given in Food Facilitates Dermatophytosis Cure in Guinea Pig Models,” Journal of Antimicrobial Chemotherapy, Vol. 46, No. 4, 2000, pp. 595-601. doi:10.1093/jac/46.4.595
[20] H. Wakabayashi, K. Yamauchi and M. Takase, “Lactoferrin Research, Technology and Applications,” International Dairy Journal, Vol. 16, No. 11, 2006, pp. 1241-1251. doi:10.1016/j.idairyj.2006.06.013
[21] M. Egashira, T. Takayanagi, M. Moriuchi and H. Moriuchi, “Does Daily Intake of Bovine Lactoferrin-Containing Products Ameliorate Rotaviral Gastroenteritis?” Acta Paediatrica, Vol. 96, No. 8, 2007, pp. 1238-1244. doi:10.1111/j.1651-2227.2007.00393.x
[22] R. Hahn, P. M. Schultz, C. Schaupp and A. Jungbauer, “Bovine Whey Fractionation Based on Cation-Exchange Chromatography,” Journal of Chromatography A, Vol. 795, No. 2, 1998, pp. 277-287. doi:10.1016/S0021-9673(97)01030-3
[23] L. Chen, C. Guo, Y. Guan and H. Liu, “Isolation of Lactoferrin from Acid Whey by Magnetic Affinity Separation,” Separation and Purification Technology, Vol. 56, No. 2, 2007, pp. 168-174. doi:10.1016/j.seppur.2007.01.019
[24] P. H. C. Van Berkel, M. E. J. Geerts, H. A. van Veen, P. M. Kooiman, H. A. Pieper, A. de Boer and J. H. Nuijens, “Glycosylated and Unglycosylated Human Lactoferrins Both Bind Iron and Show Identical Affinities towards Human Lysozyme and Bacterial Lipopolysaccharide, but Differ in Their Susceptibilities towards Tryptic Proteolysis,” Biochemical Journal, Vol. 312, Pt. 1, 1995, pp. 107-114.
[25] U. K. Laemmli, “Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4,” Nature, Vol. 227, 1970, pp. 680-685. doi:10.1038/227680a0
[26] H. A. E. Asfour, M. H. Yassin and A. M. Gomaa, “Antibacterial Activity of Bovine Milk Lactoferrin against Some Mastitis Causative Pathogens with Special Regards to Mycoplasmas,” International Journal of Microbiology Research, Vol. 1, No. 3, 2010, pp. 97-105.
[27] N. Richardson, “Standard Methods for the Examination of Dairy Product,” 15th Edition, APHA, Washington DC, 1985.
[28] G. Rai-Elbalhaa, J. L. Pellerin, G. Bodin, H. A. Abdullah and H. Hiron, “Lymphoblastic Transformation Assay of Sheep Peripheral Blood Lymphocytes: A New Rapid and Easy-to-Read Technique,” Comparative Immunology, Microbiology and Infectious Diseases, Vol. 8, 1985, pp. 311-318. doi:10.1016/0147-9571(85)90010-4
[29] M. Borenstein, H. Rothstein and J. Cohen, “Sample Power Statistics 10,” SPSS Inc., Chicago, 1997.
[30] E. Fuda, P. Jauregi and D. L. Pyle, “Recovery of Lactoferrin and Lactoperoxidase from Sweet Whey Using Colloidal Gas Aphrons (CGAs) Generated from an Anionic Surfactant,” AOT Biotechnology Progress, Vol. 20, No. 2, 2004, pp. 514-525. doi:10.1021/bp034198d
[31] R. Ghosh, “Protein Separation Using Membrane Chromatography: Opportunities and Challenges,” Journal of Chromatography A, Vol. 952, No. 1-2, 2002, pp. 13-27.
[32] N. B. Iannucci, F. J. Wolman, S. A. Camperi, A. A. N. Canizo, M. Grasselli and O. Cascone, “Affinity Chromatography with Pseudobiospecific Ligands on High-Performance Supports for Purification of Proteins of Biotechnological Interest,” Brazilian Journal of Chemical Engineering, Vol. 20, No. 1, 2003, p. 27. doi:10.1590/S0104-66322003000100006
[33] D. L. Stewart, D. Purvis and C. R. Lowe, “Affinity Chromatography on Novel Perfluorocarbon Supports: Immobilisation of C.I. Reactive Blue 2 on a Polyvinyl Alcohol-Coated Perfluoropolymer Support and Its Application in Affinity Chromatography,” Journal of Chromatography A, Vol. 510, 1990, pp. 177-187.
[34] F. J. Wolman, D. Gonzalez Maglio, M. Grasselli and O. Cascone, “One-Step Lactoferrin Purification from Bovine Whey and Colostrum by Affinity Membrane Chromatography,” Journal of Membrane Science, Vol. 288, No. 1-2, 2007, pp. 132-138. doi:10.1016/j.memsci.2006.11.011
[35] S. Yoshida and X. Y. Ye, “Isolation of Lactoperoxidase and Lactoferrin from Bovine Milk Acid Whey by Carboxymethyl Cation Exchange Chromatography,” Journal of Dairy Science, Vol. 74, No. 5, 1991, pp. 1439-1444. doi:10.3168/jds.S0022-0302(91)78301-X
[36] R. S. Bhimani, Y. Vendrov and P. Furmanski, “Influence of Lactoferrin Feeding and Injection against Systemic Staphylococcal Infections in Mice,” Journal of Applied Microbiology, Vol. 86, No. 1, 1999, pp. 135-144. doi.10.1046/j.1365-2672.1999.00644.x
[37] K. Yamauchi, M. Tomita, T. J. Giehl and R. T. Ellison III, “Antibacterial Activity of Lactoferrin and a Pepsin-Derived Lactoferrin Peptide Fragment,” Infection and Immunity, Vol. 61, No. 2, 1993, pp. 719-728.
[38] S. Farnaud and R. W. Evans, “Lactoferrin: A Multi-Functional Protein with Antimicrobial Properties,” Molecular Immunology, Vol. 40, No. 7, 2003, pp. 395-405. doi:10.1016/S0161-5890(03)00152-4
[39] P. Valenti and G. Antonini, “Lactoferrin: An Important Host Defense against Microbial and Viral Attack,” Cellular and Molecular Life Sciences, Vol. 62, No. 22, 2005, pp. 2576-2587. doi:10.1007/s00018-005-5372-0
[40] A. Nacimiento and L. O. Giugliano, “Human Milk Fractions Inhibit the Adherence of Diffusely Adherent Escherichia coli (DAEC) and enteroaggregative E. coli (EAEC) to HeLa Cells,” FEMS Microbiology Letters, Vol. 184, No. 1, 2000, pp. 91-94.
[41] P. K. Singh, M. R. Parsek, E. P. Greenberg and M. J. Welsh, “A Component of Innate Immunity Prevents Bacterial Biofilm Development,” Nature, Vol. 417, No. 6888, 2002, pp. 552-555.
[42] E. D. Weinberg, “Suppression of Bacterial Biofilm Formation by Iron Limitation,” Medical Hypotheses, Vol. 63, No. 5, 2004, pp. 863-865. doi:10.1016/j.mehy.2004.04.010
[43] A. S. Naidu, “Lactoferrin: Natural, Multifunctional, Antimicrobial,” CRC Press LLC, Boca Raton, 2000. doi:10.1201/9781420041439
[44] R. Gonzalo, D. Yang, P. Tewary, A. Varadhachary and J. J. Oppenheim, “Lactoferrin Acts as an Alarmin to Promote the Recruitment and Activation of APCs and Antigen-Specific Immune Responses,” Journal of Immunology, Vol. 180, No. 10, 2008, pp. 6868-6876.
[45] M. Zimecki, S. Dariusz, A. S. Pniak and L. K. Marian, “Lactoferrin Regulates Proliferative Response of Human Peripheral Blood Mononuclear Cells to Phytohaemaglutinin and Mixed Lymphocyte Reaction,” Archivum Immunologiae et Therapiae Experimentalis, Vol. 49, No. 2, 2001, pp. 147-154.
[46] E. R. Richie, J. K. Hilliard, R. Gilmore and D. J. Gillespie, “Human Milk Derived Lactoferrin Inhibits Mitogen and Alloantigen Induced Human Lymphocyte Proliferation,” Journal of Reproductive Immunology, Vol. 12, No. 2, 1987, pp. 137-148. doi:10.1016/0165-0378(87)90041-6
[47] H. Wakabayashi, H. Kuwata, K. Yamauchi, S. Teraguchi and Y. Tamura, “No Detectable Transfer of Dietary Lactoferrin or Its Functional Fragments to Portal Blood in Healthy Adult Rats,” Bioscience, Biotechnology, and Biochemistry, Vol. 68, No. 4, 2004, pp. 853-860. doi:10.1271/bbb.68.853
[48] S. Teraguchi, H. Wakabayashi, H. Kuwata, K. Yamauchi and Y. Tamura, “Protection against Infections by Oral Lactoferrin: Evaluation in Animal Models,” Biometals, Vol. 17, No. 3, 2004, pp. 231-234. doi:10.1023/B:BIOM.0000027697.83706.32
[49] W. P. Wang, M. Iigo, J. Sato, K. Sekine, I. Adachi and H. Tsuda, “Activation of Intestinal Mucosal Immunity in Tumor-Bearing Mice by Lactoferrin,” Cancer Science, Vol. 91, No. 10, 2000, pp. 1022-1027. doi:10.1111/j.1349-7006.2000.tb00880.x
[50] N. Takakura, H. Wakabayashi, K. Yamauchi and M. Takase, “Influences of Orally Administered Lactoferrin on IFN-γ and IL-10 Production by Intestinal Intraepithelial Lymphocytes and Mesenteric Lymph-Node Cells,” Biochemistry and Cell Biology, Vol. 84, No. 3, 2006, pp. 363-368. doi:10.1139/o06-056

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