1887

Abstract

Biofilm-associated infections are difficult to eradicate because of their ability to tolerate antibiotics and evade host immune responses. Amoebae and/or their secreted products may provide alternative strategies to inhibit and disperse biofilms on biotic and abiotic surfaces. We evaluated the potential of five predatory amoebae – , , , and – and their cell-free secretions to disrupt biofilms formed by methicillin-resistant (MRSA) and . The biofilm biomass produced by MRSA and was significantly reduced when co-incubated with , and , and their corresponding cell-free supernatants (CFS). spp. generally produced CFS that mediated biofilm dispersal rather than directly killing the bacteria; however, CFS demonstrated active killing of MRSA planktonic cells when the bacteria were present at low concentrations. The active component(s) of the CFS is resistant to freezing, but can be inactivated to differing degrees by mechanical disruption and exposure to heat. and its CFS also reduced preformed biofilms, whereas only decreased biofilm biomass when amoebae were added. These results highlight the potential of using select amoebae species or their CFS to disrupt preformed bacterial biofilms.

Funding
This study was supported by the:
  • Defense Advanced Research Projects Agency (Award W911NF-15–2-0124)
    • Principle Award Recipient: Bradley R. Borlee
  • Defense Advanced Research Projects Agency (Award W911NF-15–2-0124)
    • Principle Award Recipient: Mary Jackson
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2020-05-27
2021-07-29
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References

  1. Flemming H-C, Wingender J, Szewzyk U, Steinberg P, Rice SA et al. Biofilms: an emergent form of bacterial life. Nat Rev Microbiol 2016; 14:563–575 [View Article][PubMed]
    [Google Scholar]
  2. Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms: a common cause of persistent infections. Science 1999; 284:1318–1322 [View Article][PubMed]
    [Google Scholar]
  3. Hathroubi S, Mekni MA, Domenico P, Nguyen D, Jacques M. Biofilms: microbial shelters against antibiotics. Microb Drug Resist 2017; 23:147–156 [View Article][PubMed]
    [Google Scholar]
  4. Balcázar JL, Subirats J, Borrego CM. The role of biofilms as environmental reservoirs of antibiotic resistance. Front Microbiol 2015; 6:1216 [View Article][PubMed]
    [Google Scholar]
  5. Bernier SP, Lebeaux D, DeFrancesco AS, Valomon A, Soubigou G et al. Starvation, together with the SOS response, mediates high biofilm-specific tolerance to the fluoroquinolone ofloxacin. PLoS Genet 2013; 9:e1003144 [View Article][PubMed]
    [Google Scholar]
  6. Gilbert P, Maira-Litran T, McBain AJ, Rickard AH, Whyte FW. The physiology and collective recalcitrance of microbial biofilm communities. Adv Microb Physiol 2002; 46:202–256[PubMed]
    [Google Scholar]
  7. Paharik AE, Horswill AR. The staphylococcal biofilm: adhesins, regulation, and host response. Microbiol Spectr 2016; 4: [View Article][PubMed]
    [Google Scholar]
  8. Mekni MA, Bouchami O, Achour W, Ben Hassen A. Strong biofilm production but not adhesion virulence factors can discriminate between invasive and commensal Staphylococcus epidermidis strains. APMIS 2012; 120:605–611 [View Article][PubMed]
    [Google Scholar]
  9. Ridenhour BJ, Metzger GA, France M, Gliniewicz K, Millstein J et al. Persistence of antibiotic resistance plasmids in bacterial biofilms. Evol Appl 2017; 10:640–647 [View Article][PubMed]
    [Google Scholar]
  10. Norrby SR, Nord CE, Finch R. European Society of clinical M, infectious D. lack of development of new antimicrobial drugs: a potential serious threat to public health. Lancet Infect Dis 2005; 5:115–119
    [Google Scholar]
  11. Ghodbane R, Medie FM, Lepidi H, Nappez C, Drancourt M. Long-Term survival of tuberculosis complex mycobacteria in soil. Microbiology 2014; 160:496–501 [View Article][PubMed]
    [Google Scholar]
  12. Bhattacharya M, Berends ETM, Chan R, Schwab E, Roy S et al. Staphylococcus aureus biofilms release leukocidins to elicit extracellular trap formation and evade neutrophil-mediated killing. Proc Natl Acad Sci U S A 2018; 115:7416–7421 [View Article][PubMed]
    [Google Scholar]
  13. McDade JJ, Hall LB. Survival of Staphylococcus aureus in the environment. I. exposure of surfaces. Am J Hyg 1963; 78:330–337 [View Article][PubMed]
    [Google Scholar]
  14. Chen AF, Wessel CB, Rao N. Staphylococcus aureus screening and decolonization in orthopaedic surgery and reduction of surgical site infections. Clin Orthop Relat Res 2013; 471:2383–2399 [View Article][PubMed]
    [Google Scholar]
  15. Noskin GA, Rubin RJ, Schentag JJ, Kluytmans J, Hedblom EC et al. National trends in Staphylococcus aureus infection rates: impact on economic burden and mortality over a 6-year period (1998-2003). Clin Infect Dis 2007; 45:1132–1140 [View Article][PubMed]
    [Google Scholar]
  16. Nourbakhsh F, Namvar AE. Detection of genes involved in biofilm formation in Staphylococcus aureus isolates. GMS Hyg Infect Control 2016; 11:Doc07 [View Article][PubMed]
    [Google Scholar]
  17. Richards MJ, Edwards JR, Culver DH, Gaynes RP. Nosocomial infections in combined medical-surgical intensive care units in the United States. Infect Control Hosp Epidemiol 2000; 21:510–515 [View Article][PubMed]
    [Google Scholar]
  18. Klevens RM, Morrison MA, Nadle J, Petit S, Gershman K et al. Invasive methicillin-resistant Staphylococcus aureus infections in the United States. JAMA 2007; 298:1763–1771 [View Article][PubMed]
    [Google Scholar]
  19. Cha J-O, Yoo JI, Yoo JS, Chung H-S, Park S-H et al. Investigation of Biofilm Formation and its Association with the Molecular and Clinical Characteristics of Methicillin-resistant Staphylococcus aureus . Osong Public Health Res Perspect 2013; 4:225–232 [View Article][PubMed]
    [Google Scholar]
  20. O'Neill E, Pozzi C, Houston P, Smyth D, Humphreys H et al. Association between methicillin susceptibility and biofilm regulation in Staphylococcus aureus isolates from device-related infections. J Clin Microbiol 2007; 45:1379–1388 [View Article][PubMed]
    [Google Scholar]
  21. Yarwood JM, Bartels DJ, Volper EM, Greenberg EP. Quorum sensing in Staphylococcus aureus biofilms. J Bacteriol 2004; 186:1838–1850 [View Article][PubMed]
    [Google Scholar]
  22. Olea-Popelka F, Muwonge A, Perera A, Dean AS, Mumford E et al. Zoonotic tuberculosis in human beings caused by Mycobacterium bovis-a call for action. Lancet Infect Dis 2017; 17:e21–e5 [View Article][PubMed]
    [Google Scholar]
  23. Palmer MV. Mycobacterium bovis: characteristics of wildlife reservoir hosts. Transbound Emerg Dis 2013; 60 Suppl 1:1–13 [View Article][PubMed]
    [Google Scholar]
  24. Maddock EC. Studies on the survival time of the bovine tubercle Bacillus in soil, soil and dung, in dung and on grass, with experiments on the preliminary treatment of infected organic matter and the cultivation of the organism. J Hyg 1933; 33:103–117 [View Article][PubMed]
    [Google Scholar]
  25. Young JS, Gormley E, Wellington EMH. Molecular detection of Mycobacterium bovis and Mycobacterium bovis BCG (Pasteur) in soil. Appl Environ Microbiol 2005; 71:1946–1952 [View Article][PubMed]
    [Google Scholar]
  26. Sanchez-Hidalgo A, Obregón-Henao A, Wheat WH, Jackson M, Gonzalez-Juarrero M. Mycobacterium bovis hosted by free-living-amoebae permits their long-term persistence survival outside of host mammalian cells and remain capable of transmitting disease to mice. Environ Microbiol 2017; 19:4010-4021 [View Article][PubMed]
    [Google Scholar]
  27. Cosivi O, Grange JM, Daborn CJ, Raviglione MC, Fujikura T et al. Zoonotic tuberculosis due to Mycobacterium bovis in developing countries. Emerg Infect Dis 1998; 4:59–70 [View Article][PubMed]
    [Google Scholar]
  28. Sintayehu DW, Prins HHT, Heitkönig IMA, de Boer WF. Disease transmission in animal transfer networks. Prev Vet Med 2017; 137:36–42 [View Article][PubMed]
    [Google Scholar]
  29. Grange JM. Mycobacterium bovis infection in human beings. Tuberculosis 2001; 81:71–77 [View Article][PubMed]
    [Google Scholar]
  30. Müller B, Dürr S, Alonso S, Hattendorf J, Laisse CJM et al. Zoonotic Mycobacterium bovis-induced tuberculosis in humans. Emerg Infect Dis 2013; 19:899–908 [View Article][PubMed]
    [Google Scholar]
  31. Adetunji VO, Kehinde AO, Bolatito OK, Chen J. Biofilm formation by Mycobacterium bovis: influence of surface kind and temperatures of sanitizer treatments on biofilm control. Biomed Res Int 2014; 2014:210165 [View Article][PubMed]
    [Google Scholar]
  32. Chen M, Yu Q, Sun H. Novel strategies for the prevention and treatment of biofilm related infections. Int J Mol Sci 2013; 14:18488–18501 [View Article][PubMed]
    [Google Scholar]
  33. Landini P, Antoniani D, Burgess JG, Nijland R. Molecular mechanisms of compounds affecting bacterial biofilm formation and dispersal. Appl Microbiol Biotechnol 2010; 86:813–823 [View Article][PubMed]
    [Google Scholar]
  34. Huws SA, McBain AJ, Gilbert P. Protozoan grazing and its impact upon population dynamics in biofilm communities. J Appl Microbiol 2005; 98:238–244 [View Article][PubMed]
    [Google Scholar]
  35. Pickup ZL, Pickup R, Parry JD. A comparison of the growth and starvation responses of Acanthamoeba castellanii and Hartmannella vermiformis in the presence of suspended and attached Escherichia coli K12. FEMS Microbiol Ecol 2007; 59:556–563 [View Article][PubMed]
    [Google Scholar]
  36. Seiler C, van Velzen E, Neu TR, Gaedke U, Berendonk TU et al. Grazing resistance of bacterial biofilms: a matter of predators’ feeding trait. FEMS Microbiol Ecol 2017; 93: [View Article][PubMed]
    [Google Scholar]
  37. Rodríguez-Zaragoza S. Ecology of free-living amoebae. Crit Rev Microbiol 1994; 20:225–241 [View Article][PubMed]
    [Google Scholar]
  38. Iqbal J, Siddiqui R, Khan NA. Acanthamoeba and bacteria produce antimicrobials to target their counterpart. Parasit Vectors 2014; 7:56 [View Article][PubMed]
    [Google Scholar]
  39. Pederson K. Biofilm development on stainless steel and PVC surfaces in drinking water. Water Research 1990; 24:239–243
    [Google Scholar]
  40. Guimaraes AJ, Gomes KX, Cortines JR, Peralta JM, Peralta RHS. Acanthamoeba spp. as a universal host for pathogenic microorganisms: One bridge from environment to host virulence. Microbiol Res 2016; 193:30–38 [View Article][PubMed]
    [Google Scholar]
  41. Fouque E, Trouilhé M-C, Thomas V, Hartemann P, Rodier M-H et al. Cellular, biochemical, and molecular changes during encystment of free-living amoebae. Eukaryot Cell 2012; 11:382–387 [View Article][PubMed]
    [Google Scholar]
  42. Fouque E, Yefimova M, Trouilhé M-C, Quellard N, Fernandez B et al. Morphological Study of the Encystment and Excystment of Vermamoeba vermiformis Revealed Original Traits. J Eukaryot Microbiol 2015; 62:327–337 [View Article][PubMed]
    [Google Scholar]
  43. Marciano-Cabral F, Cabral G. Acanthamoeba spp. as agents of disease in humans. Clin Microbiol Rev 2003; 16:273–307 [View Article][PubMed]
    [Google Scholar]
  44. Booton GC, Visvesvara GS, Byers TJ, Kelly DJ, Fuerst PA. Identification and distribution of Acanthamoeba species genotypes associated with nonkeratitis infections. J Clin Microbiol 2005; 43:1689–1693 [View Article][PubMed]
    [Google Scholar]
  45. Bouchoucha I, Aziz A, Hoffart L, Drancourt M. Repertoire of free-living protozoa in contact lens solutions. BMC Ophthalmol 2016; 16:191 [View Article][PubMed]
    [Google Scholar]
  46. Salameh A, Bello N, Becker J, Zangeneh T. Fatal granulomatous amoebic encephalitis caused by Acanthamoeba in a patient with kidney transplant: a case report. Open Forum Infect Dis 2015; 2::ofv104 [View Article][PubMed]
    [Google Scholar]
  47. Lass A, Guerrero M, Li X, Karanis G, Ma L et al. Detection of Acanthamoeba spp. in water samples collected from natural water reservoirs, sewages, and pharmaceutical factory drains using LAMP and PCR in China. Sci Total Environ 2017; 584-585:489-494 [View Article][PubMed]
    [Google Scholar]
  48. el Sibae MM. Detection of free-living amoeba (Acanthamoeba polyphagia) in the air conditioning systems. J Egypt Soc Parasitol 1993; 23:687–690[PubMed]
    [Google Scholar]
  49. Fouque E, Trouilhé M-C, Thomas V, Humeau P, Héchard Y. Encystment of Vermamoeba (Hartmannella) vermiformis: Effects of environmental conditions and cell concentration. Exp Parasitol 2014; 145 Suppl:S62–68 [View Article][PubMed]
    [Google Scholar]
  50. Wheat WH, Casali AL, Thomas V, Spencer JS, Lahiri R et al. Long-term survival and virulence of Mycobacterium leprae in amoebal cysts. PLoS Negl Trop Dis 2014; 8:e3405 [View Article][PubMed]
    [Google Scholar]
  51. Pagnier I, Valles C, Raoult D, La Scola B. Isolation of Vermamoeba vermiformis and associated bacteria in hospital water. Microb Pathog 2015; 80:14–20 [View Article][PubMed]
    [Google Scholar]
  52. Hilbi H, Weber SS, Ragaz C, Nyfeler Y, Urwyler S. Environmental predators as models for bacterial pathogenesis. Environ Microbiol 2007; 9:563–575 [View Article][PubMed]
    [Google Scholar]
  53. Kessin RH. Dictyostelium: Evolution, Cell Biology, and the Development of Multicellularity 38 Cambridge University Press; 2001
    [Google Scholar]
  54. DiSalvo S, Brock DA, Smith J, Queller DC, Strassmann JE. In the social amoeba Dictyostelium discoideum, density, not farming status, determines predatory success on unpalatable Escherichia coli . BMC Microbiol 2014; 14:328 [View Article][PubMed]
    [Google Scholar]
  55. de Souza TK, Soares SS, Benitez LB, Rott MB. Interaction between methicillin-resistant Staphylococcus aureus (MRSA) and Acanthamoeba polyphaga . Curr Microbiol 2017; 74:541-549 [View Article][PubMed]
    [Google Scholar]
  56. Huws SA, Smith AW, Enright MC, Wood PJ, Brown MRW. Amoebae promote persistence of epidemic strains of MRSA. Environ Microbiol 2006; 8:1130–1133 [View Article][PubMed]
    [Google Scholar]
  57. Long JJ, Jahn CE, Sánchez-Hidalgo A, Wheat W, Jackson M et al. Interactions of free-living amoebae with rice bacterial pathogens Xanthomonas oryzae pathovars oryzae and oryzicola . PLoS One 2018; 13:e0202941 [View Article][PubMed]
    [Google Scholar]
  58. Udekwu KI, Parrish N, Ankomah P, Baquero F, Levin BR. Functional relationship between bacterial cell density and the efficacy of antibiotics. J Antimicrob Chemother 2009; 63:745–757 [View Article][PubMed]
    [Google Scholar]
  59. Upadhyay JM. Growth and bacteriolytic activity of a soil amoeba, Hartmannella glebae . J Bacteriol 1968; 95:771–774[PubMed]
    [Google Scholar]
  60. Markman DW, Antolin MF, Bowen RA, Wheat WH, Woods M et al. Yersinia pestis Survival and Replication in Potential Ameba Reservoir. Emerg Infect Dis 2018; 24:294–302 [View Article][PubMed]
    [Google Scholar]
  61. Greub G, Raoult D. Microorganisms resistant to free-living amoebae. Clin Microbiol Rev 2004; 17:413–433 [View Article][PubMed]
    [Google Scholar]
  62. El-Etr SH, Margolis JJ, Monack D, Robison RA, Cohen M et al. Francisella tularensis type A strains cause the rapid encystment of Acanthamoeba castellanii and survive in amoebal cysts for three weeks postinfection. Appl Environ Microbiol 2009; 75:7488–7500 [View Article][PubMed]
    [Google Scholar]
  63. Zheng J, Ho B, Mekalanos JJ. Genetic analysis of anti-amoebae and anti-bacterial activities of the type VI secretion system in Vibrio cholerae . PLoS One 2011; 6:e23876 [View Article][PubMed]
    [Google Scholar]
  64. Kim W-T, Kong H-H, Ha Y-R, Hong Y-C, Jeong HJ et al. Comparison of specific activity and cytopathic effects of purified 33 kDa serine proteinase from Acanthamoeba strains with different degree of virulence. Korean J Parasitol 2006; 44:321–330 [View Article][PubMed]
    [Google Scholar]
  65. Ramírez-Rico G, Martínez-Castillo M, de la Garza M, Shibayama M, Serrano-Luna J. Acanthamoeba castellanii Proteases are Capable of Degrading Iron-Binding Proteins as a Possible Mechanism of Pathogenicity. J Eukaryot Microbiol 2015; 62:614–622 [View Article][PubMed]
    [Google Scholar]
  66. Serrano-Luna J, Piña-Vázquez C, Reyes-López M, Ortiz-Estrada G, de la Garza M. Proteases from Entamoeba spp. and Pathogenic Free-Living Amoebae as Virulence Factors. J Trop Med 2013; 2013:890603 [View Article][PubMed]
    [Google Scholar]
  67. Ferreira GA, Magliano ACM, Pral EMF, Alfieri SC. Elastase secretion in Acanthamoeba polyphaga . Acta Trop 2009; 112:156–163 [View Article][PubMed]
    [Google Scholar]
  68. Leippe M. Pore-Forming toxins from pathogenic amoebae. Appl Microbiol Biotechnol 2014; 98:4347–4353 [View Article][PubMed]
    [Google Scholar]
  69. Clarke DW, Niederkorn JY. The pathophysiology of Acanthamoeba keratitis. Trends Parasitol 2006; 22:175–180 [View Article][PubMed]
    [Google Scholar]
  70. Anderson IJ, Watkins RF, Samuelson J, Spencer DF, Majoros WH et al. Gene discovery in the Acanthamoeba castellanii genome. Protist 2005; 156:203–214 [View Article][PubMed]
    [Google Scholar]
  71. Lin W-C, Tsai C-Y, Huang J-M, Wu S-R, Chu LJ et al. Quantitative proteomic analysis and functional characterization of Acanthamoeba castellanii exosome-like vesicles. Parasit Vectors 2019; 12:467 [View Article][PubMed]
    [Google Scholar]
  72. Jeyaram A, Jay SM. Preservation and storage stability of extracellular vesicles for therapeutic applications. Aaps J 2017; 20:1 [View Article][PubMed]
    [Google Scholar]
  73. Deolindo P, Evans-Osses I, Ramirez MI. Microvesicles and exosomes as vehicles between protozoan and host cell communication. Biochem Soc Trans 2013; 41:252–257 [View Article][PubMed]
    [Google Scholar]
  74. Davis SL, Perri MB, Donabedian SM, Manierski C, Singh A et al. Epidemiology and outcomes of community-associated methicillin-resistant Staphylococcus aureus infection. J Clin Microbiol 2007; 45:1705–1711 [View Article][PubMed]
    [Google Scholar]
  75. Chiu IM, Heesters BA, Ghasemlou N, Von Hehn CA, Zhao F et al. Bacteria activate sensory neurons that modulate pain and inflammation. Nature 2013; 501:52–57 [View Article][PubMed]
    [Google Scholar]
  76. Waters WR, Thacker TC, Nelson JT, DiCarlo DM, Maggioli MF et al. Virulence of two strains of Mycobacterium bovis in cattle following aerosol infection. J Comp Pathol 2014; 151:410–419 [View Article][PubMed]
    [Google Scholar]
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