Antibiofilm Activities in Skin Secretions of Malaysian Frogs

  • Authors

    • A Abdul-Aziz
    • M S Razali
    • W M S Wan Azmi
    • Z A Rahman
    • M F F Abdullah
    2019-12-24
    https://doi.org/10.14419/ijet.v7i4.14.27463
  • Antibiofilm, Peptide, Biofilm formation, Biofilm inhibition assay, Frog skin secretions.

  • Abstract

    Biofilms are extracellular structures formed by many species of bacteria that attached on various surfaces. The formation of biofilms is responsible for many health problems as they are very difficult to treat effectively due to enhanced resistant to antibiotics. Frog skin secretions contain bioactive compounds that may exhibit antibiofilms properties against biofilms infection. This study aims to screen the presence of antibiofilm activities from lyophilized skin secretions collected from Malaysian frogs. In this study, lyophilized skin secretions from four species of Malaysian frogs were tested for biofilm inhibition activities against the bacteria Staphylococcus epidermidis, ATCC 35984. Antibiofilm activity was tested at three different stages of biofilm formation which were biofilm maturation, biofilm attachment and biofilm dispersion. Biofilm treated with skin secretion from Fejervarya multistriata showed the presence of antibiofilm activities of 45 % which was able to reduce biofilm formation to 55 % at the attachment stage. Similarly, the biofilm formation at the maturation stage was reduced to 65.8 % only. No antibiofilm activities of Fejervarya multistriata was detected at the dispersion stage. The sample showing positive results was fractionated using column chromatography and further purified by C18 reverse phase high-performance liquid chromatography column (HPLC). Mass spectroscopy revealed the presence of a single peptide. In conclusion, frog skin secretion of F. multistriata contains peptide that inhibits biofilm at both attachment and maturation stages.

     

  • References

    1. [1] Costerton JW, Lewandowski Z., Caldwell DE, Korber DR & Lappin-Scott HM (1995), Microbial biofilms. Annu Rev Microbiol 49, 711–745.

      [2] Donlan RM & Costerton JW (2002), Biofilms: survival mechanisms of clinically relevant microorganisms. ClinMicrobiol Rev 15, 167–193.

      [3] Costerton JW, Stewart PS & Greenberg EP (1999), Bacterial biofilms: a common cause of persistent infections. Science 284, (5418), 1318-1322.

      [4] de la Fuente-Nu´n˜ez C, Reffuveille F, Ferna´ndez L, Hancock REW (2013), Bacterial biofilm development as a multicellular adaptation: antibiotic resistance and new therapeutic strategies. Curr Opin Microbiol 16, 580–9.

      [5] O’Neill E, Pozzi C, Houston P, Humphreys H, Robinson DA, Loughman DA, Foster TJ & O’Gara J (2008), A novel Staphylococcus aureus biofilm phenotype mediated by the fibronectin-binding proteins, FnBPA and FnBPB. J Bacteriol 190, 3835–3850.

      [6] Garrett TG, Bhakoo M & Zhang Z (2008), Bacterial adhesion and biofilms on surfaces. Prog Nat Sci 18, 1049–1056.

      [7] Francois P, Schrenzel J, Stoerman-Chopard C, Favre H, Hermann M., Foster TJ, Lew DP & Vaudaux P (2000), Identification of plasma proteins adsorbed on hemodialysis tubing that promote Staphylococcus aureus adhesion. J Lab Clin Med 135, 32–42.

      [8] Fey PD (2010), Modality of bacterial growth presents unique targets: how do we treat biofilm mediated infections?. Curr Opin Microbiol 13, 610–615.

      [9] Hoiby N, Bjarnsholt T, Givskov M, Molin S & Ciofu O (2010), Antibiotic resistance of bacterial biofilms. Int J Antimicrob Ag 35, 322–332.

      [10] Boles BR & Horswill AR (2011), Staphylococcal biofilms disassembly. Trends Microbiol 19, 449–455.

      [11] Ramlah Zainudin M &Rahman Badr MM Za (2010), Mating Calls Description of Five Species of Frogs from the Genus Hylarana tschudi 1838 (Amphibia, Anura, Ranidae) from Sarawak, Malaysia. Sains Malaysian 39(3), 363-369.

      [12] Leonardo de Azevedo Calderon, Alexandre de Almeid E Silva, Pietro Ciancaglini & Rodrigo Guerino Sta´beli (2010), Antimicrobial Peptides from Phyllomedusa frogs: from Biomolecular Diversity to Potential Nanotechnologic Medical Applications. Amino Acids DOI 10.1007/s 00726 - 010-0622-3.

      [13] Guilherme DB, Raimunda CS, Luisa Mayumi Arake, Valdelânia GS, Leiz MCV, Vladimir C, Carlos Henrique NC, Selma SK, José Guilherme Alexandre, Maria JF & José Roberto SAL (2013), The Skin Secretion of the Amphibian Phyllomedusa nordestina: A Source of Antimicrobial and Antiprotozoal Peptides. Moleculaes 18, 2058-2070.

      [14] Pukala TL, Bowie JH, Maselli VM, Musgrave IF & Tyler MJ (2006), Host-defence peptides from the glandular secretions of amphibians: structure and activity. Nat. Prod. Rep 23, 368–393.

      [15] Kückelhaus SA, Leite JRS, Muniz-Junqueira MI, Sampaio RN, Bloch C & Tosta CE (2009), Antiplasmodial and antileishmanial activities of phylloseptin-1, an antimicrobial peptide from the skin secretion of Phyllomedusa azurea (Amphibia). Exp. Parasitol 123, 11–16.

      [16] Bowie JH, Separovic F & Tyler MJ (2012), Host-defense peptides of Australian anurans. Part 2.Structure, activity, mechanism of action, and evolutionary significance. Peptides 37, 174–188.

      [17] Yeaman MR &Yount NY (2003), Mechanisms of antimicrobial peptide action and resistance. Pharmacol. Rev 55, 27–55.

      [18] Nguyen LT, Haney EF & Vogel HJ (2011), The expanding scope of antimicrobial peptide structures and their modes of action. Trends Biotechnol 29, 464–472.

      [19] Liu J, Wu Q, Li L, Xi X, Wu D, Zhou M, Chen T, Shaw C & Wang, L (2017), Discovery of phylloseptins that defense against gram-positive bacteria and inhibit the proliferation of the non-small cell lung cancer cell line, from the skin secretions of Phyllomedusa frogs. Molecues 22, 1428.

      [20] Grassi L, Maisetta G, Maccari G, Esin S & Batoni G (2017), Analogs of the frog-skin antimicrobial peptide temporin 1tb exhibit a wider spectrum of activity and a stronger antibiofilm potential as compared to the parental peptide. Front. Chem. 5.

      [21] Davies D (2003), Understanding biofilm resistance to antibacterial agents. Nat Rev Drug Discov 2(2), 114-122.

      [22] Conlon JM & Sonnevend A (2010), Antimicrobial peptides in frog skin secretions. Methods Mol. Biol. 618, 3–14.

      [23] Ma YF, Liu CB, Liu XH, Wu J, Yang HL, Wang YP, Li JX, Yu HN & Lai R (2010), Peptidomics and genomics analysis of novel antimicrobial peptides from the frog, Rana nigrovittata. Genomics 95, 66–71

      [24] da Silva BR, de Freitas VA, Carneiro VA, Arruda FV, Lorenzón EN, de Aguiar AS, Cilli EM, Cavada BS & Teixeira EH (2013), Antimicrobial activity of the synthetic peptide Lys-a1 against oral streptococci. Peptides 42, 78–83.

      [25] Shang D, Liang H, Wei S, Yan X, Yang Q, & Sun Y (2014), Effects of antimicrobial peptide L-K6, a temporin-1CEb analog on oral pathogen growth, Streptococcus mutans biofilm formation, and anti-inflammatory activity. Appl Microbiol Biotechnol 98, 8685–8695.

      [26] Luca V, Stringaro A, Colone M, Pini A & Mangoni ML (2013) Esculentin (1–21), an amphibian skin membrane-active peptide with potent activity on both planktonic and biofilm cells of the bacterial pathogen Pseudomonas aeruginosa. Cell Mol Life Sci 70, 2773–2786.

      [27] Abraham P, Sundaram A, Asha R, George S & Kumar KS (2015), Structure activity relationship and mode of action of a frog secreted antibacterial peptide B1CTcu5 using Synthetically and Modularly Modified or Deleted (SMMD) peptides. PLoS One 10, e0124210

      [28] Rahman ZA, Abdullah MFF, Wan Azmi WMS & Abdul-Aziz A (2016), Preliminary study of antimicrobial activity of the skin secretions of Malaysian frogs. Jurnal Teknologi 78(6-5), 65-69.

      [29] Wu X, Pan J, Wu Y, Xi X, Ma C, Wang L, Zhou M & Chen T (2017), PSN-PC: A Novel Antimicrobial and Anti-Biofilm Peptide from the Skin Secretion of Phyllomedusa-camba with Cytotoxicity on Human Lung Cancer Cell. Molecules 22(11),1896.

      [30] Stepanović S, Vuković D, Hola V, Di Bonaventura G, Djukić S, Cirković I & Ruzicka F (2007 ), Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. APMIS 115(8), 891–899.

      [31] He X, Yang S, Wei L, Liu R, Lai R & Rong M (2013), Antimicrobial peptide diversity in the skin of the torrent frog, Amolops jingdongensis. Amino Acids 44(2), 481–487. Published online 2012 Jul 25. doi: 10.1007/s00726-012-1358-z

      [32] Vuong C & Otto M (2002), Staphylococcus epidermidis infections. Microbes Infect 4, 481–489.

      [33] Kloos W & Schleifer KH, Bergey's Manual of Systematic Bacteriology. PHA, S.; S, M.; ME, S.; JG, H., editors, Baltimore: Williams & Wilkins, (1986).

      [34] McCann MT, Gilmore BF & Gorman SP (2008), Staphylococcus epidermidis device-related infections: pathogenesis and clinical management. J Pharm Pharmacol 60, 1551– 1571.

      [35] Rogers KL, Fey PD & Rupp ME (2009), Coagulase-negative staphylococcal infections. Infect Dis Clin North Am 23, 73–98.

      [36] Whitfield GB, Marmont LS & Howell PL (2015), Enzymatic modifications of exopolysaccharides enhance bacterial persistence. Front Microbiol 6, 471

      [37] Hassan M, Kjos M, Nes IF, Diep DB & Lotfipour F (2012), Natural antimicrobial peptides from bacteria: characteristics and potential applications to fight against antibiotic resistance. J ApplMicrobiol 113, 723–736.

      [38] Zasloff M., Martin B & Chen HC (1988), Antimicrobial activity of synthetic magainin peptides and several analogues. Proceedings of the National Academy of Sciences of the United States of America 85(3), 910–913.

      [39] Giacometti A, Cirioni O, Barchiesi F & Scalise G (2000), In-vitro activity and killing effect of polycationic peptides on methicillin-resistant Staphylococcus aureus and interactions with clinically used antibiotics. DiagnMicrobiol Infect Dis 38, 115-118.

      [40] Kaiser H, Carvalho V, Ceballos J, Freed P, Heacox S, Lester B, Richards S, Trainor C, Sanchez C & O’Shea M (2011), The herpetofauna of Timor-Leste: a first report. ZooKeys 109, 19–86

      [41] Sai KP, Jagannadham MV, Vairamani M, Raju NP, Devi AS, Nagaraj R &Sitaram N (2001), Tigerinins: novel antimicrobial peptides from the Indian frog Rana tigerina. J BiolChem 276, 2701- 07.

      [42] Yuzhu S, Yi L, Lijun W, Hailong Y, Keyun Z &Ren L (2009), Purification, characterization and cloning of two novel tigerinin-like peptides from skin secretions of Fejervarya cancrivora. Peptides 7, 1228-32.

      [43] Christensen GD, Simpson WA, Bisno AL & Beachey EH (1982), Adherence of slime-producing strains of Staphylococcus epidermidis to smooth surfaces. Infect Immun 37(1), 318–326.

      [44] Molhoek EM, van Dijk A, Veldhuizen EJ, Haagsman HP & Bikker FJ (2010), A cathelicidin-2-derived peptide effectively impairs Staphylococcus epidermidis biofilms. International Journal of Antimicrobial Agents Volume 37, Issue 5, 476 – 479

      [45] Vuong C, Voyich JM, Fisher ER, Braughton KR, Whitney AR, DeLeo, FR & Otto M (2004), Polysaccharide intercellular adhesin (PIA) protects Staphylococcus epidermidis against major components of the human innate immune system. Cell. Microbiol. 6, 269–275

      [46] Wu D, Gao Y, Tan Y, Liu Y, Wang L, Zhou M, Xi X, Ma C, Bininda-Emonds ORP, Chen T & Shaw C (2018), Discovery of Distinctin-Like-Peptide-PH (DLP-PH) From the Skin Secretion of Phyllomedusa hypochondrialis, a Prototype of a Novel Family of Antimicrobial Peptide. Frontiers in Microbiology. 9, 541.

      [47] Haney EF, Mansour SC & Hancock RE (2017), Antimicrobial Peptides: An Introduction. Methods Mol Biol. 1548, 3-22.

      [48] Andrea A, Molchanova N & Jenssen H (2018), Antibiofilm Peptides and Peptidomimetics with Focus on Surface Immobilization. Biomolecules. 8(2), 27.

      [49] Jacobsen AS & Jenssen H (2012), Human cathelicidin LL-37 prevents bacterial biofilm formation. Future Med Chem. 4(12), 1587-99.

      [50] Overhage J, Campisano A, Bains M, Torfs EC, Rehm BH, Hancock RE (2008), Human host defense peptide LL-37 prevents bacterial biofilm formation. Infect Immun. 76(9), 4176-82.

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  • How to Cite

    Abdul-Aziz, A., S Razali, M., M S Wan Azmi, W., A Rahman, Z., & F F Abdullah, M. (2019). Antibiofilm Activities in Skin Secretions of Malaysian Frogs. International Journal of Engineering & Technology, 7(4.14), 14-19. https://doi.org/10.14419/ijet.v7i4.14.27463

    Received date: 2019-02-15

    Accepted date: 2019-02-15

    Published date: 2019-12-24