Bacteriostatic and Bactericidal Activities of Camel Lactoferrins Against Salmonella enterica Serovar Typhi

  • Hussein A. Almehdar
  • Nawal Abd El-Baky
  • Abdulqader A. Alhaider
  • Saud A. Almuhaideb
  • Abdullah A. Alhaider
  • Raed S. Albiheyri
  • Vladimir N. UverskyEmail author
  • Elrashdy M. RedwanEmail author


Lactoferrin is an iron-binding glycoprotein present in various secretions (e.g., milk, tears, saliva, pancreatic juice), which performs multiple functions, with one of them being the antimicrobial defense. Purified camel lactoferrins (cLfs) from different Saudi camel clans, as well as human and bovine lactoferrins (hLf or bLf) were tested as antimicrobial agents against Salmonella enterica serovar Typhi (S. Typhi). All cLfs showed superior antibacterial potentials relative to hLf or bLf, while there was no noticeable difference in the antimicrobial capabilities between the cLfs from different camel clans. We observed synergy between the inhibitory activities of Lfs and antibiotics against bacterial growth. Expression of numerous bacterial proteins was affected by the treatment with Lf and its combinations, giving insight into the molecular mechanisms of the Lf action. Furthermore, several bacterial proteins were shown to interact with cLf-biotin. Scanning and transmission electron microscopy revealed the presence of obvious extracellular and intracellular changes after S. Typhi treatment by antibiotic (carbenicillin) or cLf alone, and in combination. The effects of antibiotics and Lf were synergistic, supporting the potential of the use of Lf-antibiotic combinations.


Antimicrobial Bovine and human lactoferrins Camel lactoferrin Salmonella Typhi Synergy 


Author Contributions

E.M.R. conceived the idea, supervised the project, organized and analyzed data, contributed to discussion, and wrote the manuscript. N.A.E.-B., H.A.A., A.A.A., S.A.A., and A.A.A. collected and analyzed data, contributed to discussion, and participated in writing of the manuscript. V.N.U. contributed to the data analysis, discussion of the results, and wrote, reviewed, and edited the manuscript.

Funding Information

This work was supported by the King Abdulaziz City for Science and Technology General Directorate of Research Grants Programs, under grant No. LGP–35–84.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no competing interests.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

12602_2019_9520_MOESM1_ESM.docx (1.4 mb)
ESM 1 (DOCX 1475 kb)


  1. 1.
    CDC (2013) Antibiotic resistance threats in the United States, 2013. Centers for Disease Control and Prevention, AtlantaGoogle Scholar
  2. 2.
    Johnson NB, Hayes LD, Brown K, Hoo EC, Ethier KA, Centers for Disease C, Prevention (2014) CDC National Health Report: leading causes of morbidity and mortality and associated behavioral risk and protective factors--United States, 2005-2013. MMWR Suppl 63(4):3–27PubMedGoogle Scholar
  3. 3.
    Ventola CL (2015) The antibiotic resistance crisis: part 2: management strategies and new agents. P T 40(5):344–352PubMedPubMedCentralGoogle Scholar
  4. 4.
    Ventola CL (2015) The antibiotic resistance crisis: part 1: causes and threats. P T 40(4):277–283PubMedPubMedCentralGoogle Scholar
  5. 5.
    ECDC, EMEA (2009) The bacterial challenge: time to react European Centre for Disease Prevention and Control European Medicines AgencyGoogle Scholar
  6. 6.
    Fish DN, Ohlinger MJ (2006) Antimicrobial resistance: factors and outcomes. Crit Care Clin 22(2):291–311 vii CrossRefGoogle Scholar
  7. 7.
    Fabrega A, Vila J (2013) Salmonella enterica serovar typhimurium skills to succeed in the host: virulence and regulation. Clin Microbiol Rev 26(2):308–341CrossRefGoogle Scholar
  8. 8.
    Buckle GC, Walker CL, Black RE (2012) Typhoid fever and paratyphoid fever: systematic review to estimate global morbidity and mortality for 2010. J Glob Health 2(1):010401CrossRefGoogle Scholar
  9. 9.
    Mogasale V, Maskery B, Ochiai RL, Lee JS, Mogasale VV, Ramani E, Kim YE, Park JK, Wierzba TF (2014) Burden of typhoid fever in low-income and middle-income countries: a systematic, literature-based update with risk-factor adjustment. Lancet Glob Health 2(10):e570–e580CrossRefGoogle Scholar
  10. 10.
    Radhakrishnan A, Als D, Mintz ED, Crump JA, Stanaway J, Breiman RF, Bhutta ZA (2018) Introductory article on global burden and epidemiology of typhoid fever. Am J Trop Med Hyg 99(3_Suppl):4–9CrossRefGoogle Scholar
  11. 11.
    Abe K, Nozaki A, Tamura K, Ikeda M, Naka K, Dansako H, Hoshino HO, Tanaka K, Kato N (2007) Tandem repeats of lactoferrin-derived anti-hepatitis C virus peptide enhance antiviral activity in cultured human hepatocytes. Microbiol Immunol 51(1):117–125CrossRefGoogle Scholar
  12. 12.
    Tatavarthy A, Luna VA, Amuso PT (2014) How multidrug resistance in typhoid fever affects treatment options. Ann N Y Acad Sci 1323:76–90CrossRefGoogle Scholar
  13. 13.
    Redwan EM, Tabll A (2007) Camel lactoferrin markedly inhibits hepatitis C virus genotype 4 infection of human peripheral blood leukocytes. J Immunoassay Immunochem 28(3):267–277CrossRefGoogle Scholar
  14. 14.
    Vitorino R (2018) Digging deep into peptidomics applied to body fluids. Proteomics 18(2).
  15. 15.
    Ganz T (2003) The role of antimicrobial peptides in innate immunity. Integr Comp Biol 43(2):300–304CrossRefGoogle Scholar
  16. 16.
    Legrand D, Pierce A, Elass E, Carpentier M, Mariller C, Mazurier J (2008) Lactoferrin structure and functions. Adv Exp Med Biol 606:163–194CrossRefGoogle Scholar
  17. 17.
    Lonnerdal B (2009) Nutritional roles of lactoferrin. Curr Opin Clin Nutr Metab Care 12(3):293–297CrossRefGoogle Scholar
  18. 18.
    Conesa C, Sanchez L, Rota C, Perez MD, Calvo M, Farnaud S, Evans RW (2008) Isolation of lactoferrin from milk of different species: calorimetric and antimicrobial studies. Comp Biochem Physiol B Biochem Mol Biol 150(1):131–139CrossRefGoogle Scholar
  19. 19.
    El-Agamy EI (2006) Camel milk. In: Park YW, Haenlan GFW (eds) Handbook of milk of non-bovine mammals. Wiley-Blackwell, HobokenGoogle Scholar
  20. 20.
    El-Fakharany EM, Abedelbaky N, Haroun BM, Sanchez L, Redwan NA, Redwan EM (2012) Anti-infectivity of camel polyclonal antibodies against hepatitis C virus in Huh7.5 hepatoma. Virol J 9:201CrossRefGoogle Scholar
  21. 21.
    Liao Y, El-Fakkarany E, Lonnerdal B, Redwan EM (2012) Inhibitory effects of native and recombinant full-length camel lactoferrin and its N and C lobes on hepatitis C virus infection of Huh7.5 cells. J Med Microbiol 61(Pt 3):375–383CrossRefGoogle Scholar
  22. 22.
    Conesa C, Calvo M, Sanchez L (2010) Recombinant human lactoferrin: a valuable protein for pharmaceutical products and functional foods. Biotechnol Adv 28(6):831–838CrossRefGoogle Scholar
  23. 23.
    Redwan EM, Uversky VN, El-Fakharany EM, Al-Mehdar H (2014) Potential lactoferrin activity against pathogenic viruses. C R Biol 337(10):581–595CrossRefGoogle Scholar
  24. 24.
    Almahdy O, El-Fakharany EM, El-Dabaa E, Ng TB, Redwan EM (2011) Examination of the activity of camel milk casein against hepatitis C virus (genotype-4a) and its apoptotic potential in hepatoma and hela cell lines. Hepat Mon 11(9):724–730CrossRefGoogle Scholar
  25. 25.
    El-Fakharany EM, El-Baky NA, Linjawi MH, Aljaddawi AA, Saleem TH, Nassar AY, Osman A, Redwan EM (2017) Influence of camel milk on the hepatitis C virus burden of infected patients. Exp Ther Med 13(4):1313–1320CrossRefGoogle Scholar
  26. 26.
    El-Fakharany EM, Haroun BM, Ng TB, Redwan ER (2010) Oyster mushroom laccase inhibits hepatitis C virus entry into peripheral blood cells and hepatoma cells. Protein Pept Lett 17(8):1031–1039CrossRefGoogle Scholar
  27. 27.
    El-Fakharany EM, Serour EA, Abdelrahman AM, Haroun BM, Redwan el RM (2009) Purification and characterization of camel (Camelus dromedarius) milk amylase. Prep Biochem Biotechnol 39(2):105–123CrossRefGoogle Scholar
  28. 28.
    El-Fakharany EM, Tabll A, Wahab AA, Redwan EM (2008) Potential activity of camel milk-amylase and lactoferrin against hepatitis C virus infectivity in HepG2 and lymphocytes. Hepat Mon 8(2):101–109Google Scholar
  29. 29.
    Redwan EM (2009) Animal-derived pharmaceutical proteins. J Immunoassay Immunochem 30(3):262–290CrossRefGoogle Scholar
  30. 30.
    el Agamy EI, Ruppanner R, Ismail A, Champagne CP, Assaf R (1992) Antibacterial and antiviral activity of camel milk protective proteins. J Dairy Res 59(2):169–175CrossRefGoogle Scholar
  31. 31.
    Redwan EM, El-Baky NA, Al-Hejin AM, Baeshen MN, Almehdar HA, Elsaway A, Gomaa AB, Al-Masaudi SB, Al-Fassi FA, AbuZeid IE, Uversky VN (2016) Significant antibacterial activity and synergistic effects of camel lactoferrin with antibiotics against methicillin-resistant Staphylococcus aureus (MRSA). Res Microbiol 167(6):480–491CrossRefGoogle Scholar
  32. 32.
    Redwan EM, El-Fakharany EM, Uversky VN, Linjawi MH (2014) Screening the anti infectivity potentials of native N- and C-lobes derived from the camel lactoferrin against hepatitis C virus. BMC Complement Altern Med 14:219CrossRefGoogle Scholar
  33. 33.
    Redwan EM, Larsen NA, Wilson IA (2003) Simplified procedure for elimination of co-purified contaminant proteins from human colostrums IgA1. J Egypt Ger Soc Zool 40(A):251–260Google Scholar
  34. 34.
    Elass-Rochard E, Roseanu A, Legrand D, Trif M, Salmon V, Motas C, Montreuil J, Spik G (1995) Lactoferrin-lipopolysaccharide interaction: involvement of the 28-34 loop region of human lactoferrin in the high-affinity binding to Escherichia coli 055B5 lipopolysaccharide. Biochem J 312(Pt 3):839–845CrossRefGoogle Scholar
  35. 35.
    Cockerill FR, Wikler MA, Alder J, Dudley MN, Eliopoulos GM, Ferraro MJ, Hardy DJ, Hecht DW, Hindler JA, Patel JB, Powell M, Swenson JM, Thomson RB, Traczewski MM, Turnidge JD, Weinstein MP, Zimmer BL (2012) Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically; Approved Standard. CLSI document M07-A9.9, vol 32, 9th edn. C.L.S.I. (Clinical and Laboratory Standard Institute), PennsylvaniaGoogle Scholar
  36. 36.
    Wikler MA (2008) Performance Standards for Antimicrobial Susceptibility Testing. Eighteenth Informational Supplement. vol 28. C.L.S.I. (Clinical and Laboratory Standard Institute), PennsylvaniaGoogle Scholar
  37. 37.
    Reddy SB, Mainwaring DE, Kobaisi MA, Zeephongsekul P, Fecondo JV (2012) Acoustic wave immunosensing of a meningococcal antigen using gold nanoparticle-enhanced mass sensitivity. Biosens Bioelectron 31(1):382–387CrossRefGoogle Scholar
  38. 38.
    Almeida RA, Luther DA, Park HM, Oliver SP (2006) Identification, isolation, and partial characterization of a novel streptococcus uberis adhesion molecule (SUAM). Vet Microbiol 115(1–3):183–191CrossRefGoogle Scholar
  39. 39.
    Sitohy MZ, Mahgoub SA, Osman AO (2012) In vitro and in situ antimicrobial action and mechanism of glycinin and its basic subunit. Int J Food Microbiol 154(1–2):19–29CrossRefGoogle Scholar
  40. 40.
    Diarra MS, Lacasse P, Deschênes E, Grondin G, Paradis-Bleau C, Petitclerc D (2003) Ultrastructural and cytochemical study of cell wall modification by lactoferrin, lactoferricin and penicillin G against Staphylococcus aureus. J Electron Microsc 52(2):207–215CrossRefGoogle Scholar
  41. 41.
    Yekta MA, Cox E, Goddeeris BM, Vanrompay D (2011) Reduction of Escherichia coli O157:H7 excretion in sheep by oral lactoferrin administration. Vet Microbiol 150(3–4):373–378CrossRefGoogle Scholar
  42. 42.
    Chapple DS, Hussain R, Joannou CL, Hancock RE, Odell E, Evans RW, Siligardi G (2004) Structure and association of human lactoferrin peptides with Escherichia coli lipopolysaccharide. Antimicrob Agents Chemother 48(6):2190–2198CrossRefGoogle Scholar
  43. 43.
    Sharma AK, Paramasivam M, Srinivasan A, Yadav MP, Singh TP (1999) Three-dimensional structure of mare diferric lactoferrin at 2.6 a resolution. J Mol Biol 289(2):303–317CrossRefGoogle Scholar
  44. 44.
    Sharma S, Sinha M, Kaushik S, Kaur P, Singh TP (2013) C-lobe of lactoferrin: the whole story of the half-molecule. Biochem Res Int 2013:271641CrossRefGoogle Scholar
  45. 45.
    Sinha M, Kaushik S, Kaur P, Sharma S, Singh TP (2013) Antimicrobial lactoferrin peptides: the hidden players in the protective function of a multifunctional protein. Int J Pept 2013:390230CrossRefGoogle Scholar
  46. 46.
    Luo G, Spellberg B, Gebremariam T, Lee H, Xiong YQ, French SW, Bayer A, Ibrahim AS (2014) Combination therapy with iron chelation and vancomycin in treating murine staphylococcemia. Eur J Clin Microbiol Infect Dis 33(5):845–851CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Biological Sciences, Faculty of SciencesKing Abdulaziz UniversityJeddahSaudi Arabia
  2. 2.Therapeutic and Protective Proteins Laboratory, Protein Research Department, Genetic Engineering and Biotechnology Research InstituteCity for Scientific Research and Technology ApplicationsNew Borg EL-ArabEgypt
  3. 3.Tilad Veterinary CenterRiyadhSaudi Arabia
  4. 4.Medical schoolKing Saud bin Abdulaziz University for health ScienceRiyadhSaudi Arabia
  5. 5.Institute for Biological Instrumentation of the Russian Academy of SciencesPushchinoRussia
  6. 6.Department of Molecular Medicine and USF Health Byrd Alzheimer’s Research Institute, Morsani College of MedicineUniversity of South FloridaTampaUSA

Personalised recommendations