Advertisement

Potential Properties of Lactobacillus plantarum F-10 as a Bio-control Strategy for Wound Infections

  • Tugce Onbas
  • Ozlem Osmanagaoglu
  • Fadime Kiran
Article
  • 22 Downloads

Abstract

In this study, Lactobacillus plantarum F-10, a promising probiotic strain isolated from fecal microbiota of healthy breastfed infant, was assessed as a bio-control strategy for wound infections. Pseudomonas aeruginosa PAO1/ATCC 27853, methicillin-resistant Staphylococcus aureus ATCC 43300, and their hospital-derived strains isolated from skin chronic wound samples were used as important skin pathogens. The cell-free extract (CFE) of the strain F-10 was shown to inhibit the growth of all pathogens tested, while no inhibition was observed when CFE was neutralized. The strain displayed no hemolysis and exhibited a strong auto-aggregating phenotype (51.48 ± 1.45%, 5 h) as well as co-aggregation. Antibiotic resistance profile was found to be safe according to the European Food Safety Authority. Biofilm formation was measured by crystal violet assay and visualized with scanning electron microscopy and confocal laser scanning microscopy. One hundred percent reduction in biofilm formation of all pathogens tested was obtained by sub-MIC value (12.5 mg/ml) of CFE following 24-h co-incubation. Inhibition of quorum sensing–controlled virulence factors (motility, protease and elastase activity, production of pyocyanin and rhamnolipid) of P. aeruginosa strains was also observed. DPPH radical scavenging activity of the CFE was determined as 88.57 ± 0.49%. In conclusion, our results suggest that L. plantarum F-10 may represent an alternative bio-control strategy against skin infections with its antimicrobial, anti-biofilm, anti-quorum sensing, and antioxidant activity.

Keywords

Lactobacillus plantarum Wound infection Probiotic Biofilm Quorum sensing Antioxidant 

Notes

Acknowledgments

The authors wish to sincerely thank Dr. Basar KARACA (Ankara University) for his excellent technical assistance on biofilm analysis.

Funding Information

This work was supported by grants of the Scientific Research Project Coordination Unit of Ankara University, Turkey (Project No: 17B0430001).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Greatrex-White S, Moxey H (2015) Wound assessment tools and nurses’ needs: an evaluation study. Int Wound J 12:293–301CrossRefPubMedGoogle Scholar
  2. 2.
    Moore Z (2012) The war on wounds. Public Sect Rev Eur 24:1–2Google Scholar
  3. 3.
    Posnett J, Gottrup F, Lundgren H et al (2009) The resource impact of wounds on health-care providers in Europe. J Wound Care 18(4):154–161CrossRefPubMedGoogle Scholar
  4. 4.
    Sen CK, Gordillo GM, Roy S et al (2009) Human skin wounds: a major and snowballing threat to public health and the economy. Wound Repair Regen 17(6):763–771CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Cogen AL, Nizet V, Gallo RL (2008) Skin microbiota: a source of disease or defence? Br J Dermatol 158:442–455CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Moore Z, Butcher G, Corbett LQ et al (2014) AAWC, AWMA, EWMA position paper: managing wounds as a team. J Wound Care 23(5):1–38Google Scholar
  7. 7.
    Serra R, Grande R, Butrico L et al (2015) Chronic wound infections: the role of Pseudomonas aeruginosa and Staphylococcus aureus. Expert Rev Anti-Infect Ther 13(5):605–613CrossRefPubMedGoogle Scholar
  8. 8.
    Plata K, Rosato AE, Wegrzyn G (2009) Staphylococcus aureus as an infectious agent: overview of biochemistry and molecular genetics of its pathogenicity. Acta Biochim Pol 56:597–612PubMedGoogle Scholar
  9. 9.
    Okhiria OA, Henriques AFM, Burton NF et al (2009) Honey modulates biofilms of Pseudomonas aeruginosa in a time and dose dependent manner. J Api Prod ApiMed Sci 1(1):6–10CrossRefGoogle Scholar
  10. 10.
    Howard JC, Reid G, Gan BS (2004) Probiotics in surgical wound infections: current status. Clin Invest Med 27(5):274–281PubMedGoogle Scholar
  11. 11.
    Markowiak P, Slizewska K (2017) Effects of probiotics, prebiotics, and synbiotics on human health. Nutrients.  https://doi.org/10.3390/nu9091021
  12. 12.
    Cinque B et al (2011) Use of probiotics for dermal applications. In: Liong MT (ed) Probiotics, Microbiology monographs. Springer-Verlag, Berlin, pp 221–241Google Scholar
  13. 13.
    Maragkoudakis PA, Zoumpopoulou G, Miaris C, Kalantzopoulos G, Pot B, Tsakalidou E (2006) Probiotic potential of Lactobacillus strains isolated from dairy products. Int Dairy J 16(3):189–199CrossRefGoogle Scholar
  14. 14.
    Wilkins TD, Holdeman LV, Abramson IJ, Moore WEC (1972) Standardized single-disc method for antibiotic susceptibility testing of anaerobic bacteria. Antimicrob Agents Chemother 1:451–459CrossRefPubMedCentralGoogle Scholar
  15. 15.
    Clinical Laboratory Standards Institute (2012) Performance standards for antimicrobial susceptibility testing; 22nd informational supplement. Document M100-S22. Clinical Laboratory Standards Institute Wayne, PAGoogle Scholar
  16. 16.
    Anonymous (2008) European food safety authority. Update of the criteria used in the assessment of bacterial resistance to antibiotics of human or veterinary importance. EFSA J 732:715Google Scholar
  17. 17.
    Savadogo A, Ouattara CAT, Bassole HNI, Traore AS (2004) Antimicrobial activities of lactic acid bacteria strains isolated from Burkina faso fermented milk. Pak J Nutr 3:174–179CrossRefGoogle Scholar
  18. 18.
    Bhunia AK, Johnson MC, Ray B (1988) Purification, characterization and antimicrobial spectrum of a bacteriocin produced by Pediococcus acidilactici. J Appl Bacteriol 65(4):261–268CrossRefPubMedGoogle Scholar
  19. 19.
    Tallon R, Bressollier P, Urdaci MC (2003) Isolation and characterization of two exopolysaccharides produced by Lactobacillus plantarum EP56. Res Microbiol 154:705–712CrossRefPubMedGoogle Scholar
  20. 20.
    Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28(3):350–356CrossRefGoogle Scholar
  21. 21.
    Del Re B, Sgorbati B, Miglioli M, Palenzona D (2000) Adhesion, autoaggregation and hydrophobicity of 13 strains of Bifidobacterium longum. Lett Appl Microbiol 31:438–442CrossRefPubMedGoogle Scholar
  22. 22.
    Kos B, Suskovic J, Vukovic S, Simpraga M, Frece J, Matosic S (2003) Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92. J Appl Microbiol 94:981–987CrossRefPubMedGoogle Scholar
  23. 23.
    Kiran F, Mokrani M, Osmanagaoglu O (2015) Effect of encapsulation on viability of Pediococcus pentosaceus OZF during its passage through the gastrointestinal tract model. Curr Microbiol 71(1):95–105CrossRefPubMedGoogle Scholar
  24. 24.
    Stepanovic S, Vukovic D, Dakic I, Savic B, Svabic-Vlahovic M (2000) A modified microtiter-plate test for quantification of staphylococcal biofilm formation. J Microbiol Methods 40(2):175–179CrossRefPubMedGoogle Scholar
  25. 25.
    Vestby LK, Moretro T, Langsrud S, Heir E, Nesse LL (2009) Biofilm forming abilities of Salmonella are correlated with persistence in fish meal and feed factories. BMC Vet Res 5(20):1–6Google Scholar
  26. 26.
    Giaouris E, Chorianopoulos N, Nychas GJE (2005) Effect of temperature, pH, and water activity on biofilm formation by Salmonella enterica Enteritidis PT4 on stainless steel surfaces as indicated by the bead vortexing method and conductance measurements. J Food Prot 68(10):2149–2154CrossRefPubMedGoogle Scholar
  27. 27.
    Rashid MH, Kornberg A (2000) Inorganic polyphosphate is needed for swimming, swarming, and twitching motilities of Pseudomonas aeruginosa. Proc Natl Acad Sci U S A 97:4885–4890CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Ramos AN, Sesto Cabral M, Noseda D, Bosch A, Yantorno OM, Valdez JC (2012) Antipathogenic properties of Lactobacillus plantarum on Pseudomonas aeruginosa: the potential use of its supernatants in the treatment of infected chronic wounds. Wound Repair Regen 20:552–562PubMedGoogle Scholar
  29. 29.
    Siegmund I, Wagner F (1991) New method for detecting rhamnolipids excreted by Pseudomonas species during growth on mineral agar. Biotechnol Tech 5:265–268CrossRefGoogle Scholar
  30. 30.
    Dong YH, Zhang LH (2005) Quorum sensing and quorum-quenching enzymes. J Microbiol 43:101–109PubMedGoogle Scholar
  31. 31.
    Yin H, Deng Y, Wang H, Liu W, Zhuang X, Chu W (2015) Tea polyphenols as an antivirulence compound disrupt quorum-sensing regulated pathogenicity of Pseudomonas aeruginosa. Nat Sci Rep 5:16158–16170CrossRefGoogle Scholar
  32. 32.
    Singleton VL, Rossi JA (1965) Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagent. J Enol Vitic 16(3):144–158Google Scholar
  33. 33.
    Zhishen J, Mengcheng T, Jianming W (1999) The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem 64(4):555–559CrossRefGoogle Scholar
  34. 34.
    Blois MS (1958) Antioxidant determinations by the use of a stable free radical. Nature 181:1199–1200CrossRefGoogle Scholar
  35. 35.
    World Health Organization (2015) Global action plan on antimicrobial resistance. WHO:1–28Google Scholar
  36. 36.
    Freedman M, Stassen LF (2013) Commonly used topical oral wound dressing materials in dental and surgical practice–a literature review. J Ir Dent Assoc 59(4):190–195PubMedGoogle Scholar
  37. 37.
    Gan BS, Kim J, Reid G, Cadieux P, Howard JC (2002) Lactobacillus fermentum RC-14 inhibits Staphylococcus aureus infection of surgical implants in rats. J Infect Dis 185:1369–1372CrossRefPubMedGoogle Scholar
  38. 38.
    Hasslöf P, Hedberg M, Twetman S, Stecksen-Blicks C (2010) Growth inhibition of oral mutans streptococci and candida by commercial probiotic lactobacilli - an in vitro study. BMC Oral Health 10:355–362CrossRefGoogle Scholar
  39. 39.
    Karska-Wysocki B, Bazo M, Smoragiewicz W (2010) Antibacterial activity of Lactobacillus acidophilus and Lactobacillus casei against methicillin-resistant Staphylococcus aureus (MRSA). Microbiol Res 165:674–686CrossRefPubMedGoogle Scholar
  40. 40.
    Peral MC, Huaman Martinez MA, Valdez JC (2009) Bacteriotherapy with Lactobacillus plantarum in burns. Int Wound J 6:73–81CrossRefPubMedGoogle Scholar
  41. 41.
    Prince T, Mcbain AJ, O’Neill CA (2012) Lactobacillus reuteri protects epidermal keratinocytes from Staphylococcus aureus-induced cell death by competitive exclusion. Appl Environ Microbiol 78:5119–5126CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Sullivan M, Schnittger SF, Mammone T, Goyarts EC (2009) Skin treatment method with Lactobacillus extract. US Patent 2005/0196480 A1. 11/070:810Google Scholar
  43. 43.
    Valdez JC, Peral MC, Rachid M, Santana M, Perdigon G (2005) Interference of Lactobacillus plantarum with Pseudomonas aeruginosa in vitro and in infected burns: the potential use of probiotics in wound treatment. Clin Microbiol Infect 11:472–479CrossRefPubMedGoogle Scholar
  44. 44.
    Al Kassaa I, Hamze M, Hober D, Chihib NE, Drider D (2014) Identification of vaginal lactobacilli with potential probiotic properties isolated from women in north Lebanon. Microb Ecol 67:722–734CrossRefPubMedGoogle Scholar
  45. 45.
    Tejero-Sarinena S, Barlow J, Costabile A, Gibson GR, Rowland I (2012) In vitro evaluation of the antimicrobial activity of a range of probiotics against pathogens: evidence for the effects of organic acids. Anaerobe 18:530–538CrossRefPubMedGoogle Scholar
  46. 46.
    Melo TA, dos Santos TF, de Almeida ME et al (2016) Inhibition of Staphylococcus aureus biofilm by Lactobacillus isolated from fine cocoa. BMC Microbiol 16:250–259CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    Simark-Mattsson C, Jonsson R, Emilson CG, Roos K (2009) Final pH affects the interference capacity of naturally occurring oral Lactobacillus strains against mutans streptococci. Arch Oral Biol 54:602–607CrossRefPubMedGoogle Scholar
  48. 48.
    Rasmussen TB, Givskov M (2006) Quorum-sensing inhibitors as anti-pathogenic drugs. Int J Med Microbiol 296:149–161CrossRefPubMedGoogle Scholar
  49. 49.
    Sadowska B, Walencka E, Wieckowska-Szakiel M et al (2010) Bacteria competing with the adhesion and biofilm formation by Staphylococcus aureus. Folia Microbiol (Praha) 55(5):497–501CrossRefGoogle Scholar
  50. 50.
    Söderling EM, Marttinen AM, Haukioja AL (2011) Probiotic lactobacilli interfere with Streptococcus mutans biofilm formation in vitro. Curr Microbiol 62(2):618–622CrossRefPubMedGoogle Scholar
  51. 51.
    Keller MK, Hasslöf P, Stecksen-Blicks C (2011) Co-aggregation and growth inhibition of probiotic lactobacilli and clinical isolates of mutans streptococci: an in vitro study. Acta Odontol Scand 69(5):263–268CrossRefPubMedGoogle Scholar
  52. 52.
    Hor YY, Liong MT (2014) Use of extracellular extracts of lactic acid bacteria and bifidobacteria for the inhibition of dermatological pathogen Staphylococcus aureus. Dermatol Sin 32:141–147CrossRefGoogle Scholar
  53. 53.
    Zmantar T, Kouidhi B, Miladi H, Mahdouani K, Bakhrouf A (2010) A microtiter plate assay for Staphylococcus aureus biofilm quantification at various pH levels and hydrogen peroxide supplementation. New Microbiol 33:137–145PubMedGoogle Scholar
  54. 54.
    Kanmani P, Kumar SR, Yuvaraj N, Paari KA et al (2011) Production and purification of a novel exopolysaccharide from lactic acid bacterium Streptococcus phocae PI80 and its functional characteristics activity in vitro. Bioresour Technol 102(7):4827–4833CrossRefPubMedGoogle Scholar
  55. 55.
    Bulgasem Y, Hassan Z, Abdalsadiq NKA, Yusoff et al (2015) Antiadhesion activity of lactic acid bacteria supernatant against human pathogenic Candida species biofilm. Health Sci J 9:1–9Google Scholar
  56. 56.
    Walencka E, Rozalska S, Sadowska B et al (2008) The influence of Lactobacillus acidophilus-derived surfactants on staphylococcal adhesion and biofilm formation. Folia Microbiol (Praha) 53(1):61–66CrossRefGoogle Scholar
  57. 57.
    Varma P, Nisha N, Dinesh KR et al (2011) Anti-infective properties of Lactobacillus fermentum against Staphylococcus aureus and Pseudomonas aeruginosa. J Mol Microbiol Biotechnol 20(3):137–143CrossRefPubMedGoogle Scholar
  58. 58.
    Ashraf R, Shah NP (2011) Antibiotic resistance of probiotic organisms and safety of probiotic dairy products. Int Food Res J 18(3):837–853Google Scholar
  59. 59.
    Mathur S, Singh R (2005) Antibiotic resistance in food lactic acid bacteria-a review. Int J Food Microbiol 105:281–295CrossRefPubMedGoogle Scholar
  60. 60.
    Liu C, Pan T (2010) in vitro effects of lactic acid bacteria on cancer cell viability and antioxidant activity. J Food Drug Anal 18:77–86Google Scholar
  61. 61.
    Zhang S, Liu L, Su Y, Li H, Sun Q, Liang X (2011) Antioxidative activity of lactic acid bacteria in yogurt. Afr J Microbiol Res 5:5194–5201Google Scholar
  62. 62.
    Xing J, Wang G, Zhang Q et al (2015) Determining antioxidant activities of lactobacilli cell-free supernatants by cellular antioxidant assay: a comparison with traditional methods. PLOS ONE.  https://doi.org/10.1371/journal.pone.0119058
  63. 63.
    Uugantsetseg E, Batjargal B (2014) Antioxidant activity of probiotic lactic acid bacteria isolated from Mongolian airag. Mongolian J Chem 15:73–78CrossRefGoogle Scholar
  64. 64.
    Pieniz S, Andreazza R, Okeke BC, Camargo FAO, Brandelli A (2015) Antimicrobial and antioxidant activities of Enterococcus species isolated from meat and dairy products. Braz J Biol.  https://doi.org/10.1590/1519-6984.02814
  65. 65.
    Wang Y, Wu Y, Wang Y et al (2017) Antioxidant properties of probiotic bacteria. Nutrients 9:521–236CrossRefPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Faculty of Science, Department of BiologyAnkara UniversityAnkaraTurkey

Personalised recommendations