Identification and characterization of Staphylococcus spp. and their susceptibility to insect apolipophorin III
Abstract
Aim: This study investigated the effect of an insect antimicrobial protein, apolipophorin III (apoLp-III), against two newly isolated, identified and characterized clinical strains of Staphylococcus spp. Materials & methods: Both strains were identified by 16S rRNA sequencing and metabolic and phenotypic profiling. The antibacterial activity of apoLp-III was tested using a colony counting assay. ApoLp-III interaction with bacterial cell surface was analyzed by Fourier transform IR spectroscopy. Results:Staphylococcus epidermidis and Staphylococcus capitis were identified. ApoLp-III exerted a dose-dependent bactericidal effect on the tested strains. The differences in the Staphylococcus spp. surface components may contribute to the various sensitivities of these strains to apoLp-III. Conclusion: ApoLp-III may provide a baseline for development of antibacterial preparations against Staphylococcus spp. involved in dermatological problems.
Papers of special note have been highlighted as: • of interest; •• of considerable interest
References
- 1. . Growing of Staphylococcus aureus cells with soil components enhances virulence in mice caused by soft tissue infections. Int. J. Pharma Bio Sci. Spl. Ed. (Int-BIONANO 2016) Conference Proceedings (ISSN: 0975-6299) 230–236 (2016).
- 2. . Screening of different soil sources of New York City for antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA). J. Pure Appl. Microbiol. 9(2), 77–86 (2015).
- 3. . Waves of resistance: staphylococcus aureus in the antibiotic era. Nat. Rev. Microbiol. 7(9), 629–641 (2009).
- 4. . Management of complications associated with totally implantable ports in patients with AIDS. AIDS Patient Care STDS 15(1), 7–13 (2001).
- 5. . Evaluation of nosocomial infection in patients at hematology-oncology ward of Dr. Sheikh Children's Hospital. Iran. J. Ped. Hematol. Oncol. 5(4), 179–185 (2015).
- 6. . Strategies for combating bacterial biofilm infections. Int. J. Oral Sci. 7(1), 1–7 (2014).
- 7. Pathogenic mechanisms and host interactions in Staphylococcus epidermidis device-related infection. Front. Microbiol. 8(1401), 1–24 (2017).
- 8. . Coagulase-negative staphylococci. Clin. Microbiol. Rev. 27(4), 870–926 (2014).
- 9. Molecular epidemiology of a vancomycin-intermediate heteroresistant Staphylococcus epidermidis outbreak in a neonatal intensive care unit. Antimicrob. Agents Chemother. 60(10), 5673–5681 (2016).
- 10. . Molecular epidemiology of Staphylococcus epidermidis clinical isolates from U.S. hospitals. Antimicrob. Agents Chemother. 56(9), 4656–4661 (2012).
- 11. . Antibiotic susceptibility and mecA frequency in Staphylococcus epidermidis, isolated from Intensive Care Unit patients. Jundishapur J. Microbiol. 7(8), e11188 (2014).
- 12. Methicillin-resistant coagulase-negative staphylococci in the community: high homology of SCCmec IVa between Staphylococcus epidermidis and major clones of methicillin-resistant Staphylococcus aureus. J. Infect. Dis. 202(2), 270–281 (2010).
- 13. . A multidrug-resistant Staphylococcus epidermidis clone (ST2) is an ongoing cause of hospital-acquired infection in a Western Australian Hospital. J. Clin. Microbiol. 50(6), 2147–2151 (2012).
- 14. . Immunity of the greater wax moth Galleria mellonella. Insect Sci. 24(3), 342–357 (2017).
- 15. Galleria mellonella as a consolidated in vivo model hosts: new developments in antibacterial strategies and novel drug testing. Virulence 10(1), 527–541 (2019). •• Summarizes the existing knowledge about use of Galleria mellonella larvae as alternative model host.
- 16. Galleria mellonella as an experimental model to study human oral pathogens. Arch. Oral Biol. 101, 13–22 (2019).
- 17. Using the wax moth larva Galleria mellonella infection model to detect emerging bacterial pathogens. PeerJ 6, e6150 (2019).
- 18. . Utilization of Galleria mellonella larvae to characterize the development of Staphylococcus aureus infection. Microbiology 165(8), 863–875 (2019).
- 19. . Insect immune activation by apolipophorin III is correlated with the lipid-binding properties of this protein. Biochemistry 40(38), 11502–11508 (2001).
- 20. . Apolipophorin III interaction with model membranes composed of phosphatidylcholine and sphingomyelin using differential scanning calorimetry. Biochim. Biophys. Acta 1788(10), 2160–2168 (2009).
- 21. . Structure of apolipophorin III in discoidal lipoproteins. Interhelical distances in the lipid-bound state and conformational change upon binding to lipid. J. Biol. Chem. 277(22), 19773–19782 (2002).
- 22. . Lipid triggered molecular switch of apoLp-III helix bundle to an extended helix conformation. J. Mol. Biol. 321(2), 201–214 (2002).
- 23. . Alternative lipid mobilization: the insect shuttle system. Mol. Cell. Biochem. 239(1–2), 113–119 (2002).
- 24. . Apolipophorin III: role model apolipoprotein. Insect. Biochem. Mol. Biol. 36(4), 231–240 (2006).
- 25. . Apolipophorins and insects immune response. Invert. Surviv. J. 10(1), 58–68 (2013). •• Describes the role of apolipophorins, with a special emphasis on apolipophorin III, in insect immune response.
- 26. . A novel role for an insect apolipoprotein (apolipophorin III) in beta-1,3-glucan pattern recognition and cellular encapsulation reactions. J. Immunol. 172(4), 2177–2185 (2004).
- 27. . An atomic force microscopy study of Galleria mellonella apolipophorin III effect on bacteria. Biochim. Biophys. Acta 1808(7), 1896–1906 (2011).
- 28. . Characterization of the apoLp-III/LPS complex: insight into the mode of binding interaction. Biochemistry 51(31), 6220–6227 (2012).
- 29. . Apolipophorin-III and the interactions of lipoteichoic acids with the immediate immune responses of Galleria mellonella. J. Invertebr. Pathol. 76, 233–241 (2000).
- 30. . Apolipophorin III: lipopolysaccharide binding requires helix bundle opening. Biochem. Biophys. Res. Commun. 348(4), 1328–1333 (2006).
- 31. . Lipopolysaccharide binding of an exchangeable apolipoprotein, apolipophorin III, from Galleria mellonella. Biol. Chem. 385(11), 1113–1119 (2004).
- 32. . Apolipophorin-III affects the activity of the haemocytes of Galleria mellonella larvae. J. Insect Physiol. 48(7), 715–723 (2002).
- 33. . Isolated apolipophorin III from Galleria mellonella stimulates the immune reactions of this insect. J. Insect Physiol. 43(4), 383–391 (1997).
- 34. . Apolipophorin III in Galleria mellonella potentiates hemolymph lytic activity. Dev. Comp. Immunol. 23(7–8), 563–570 (1999).
- 35. . Insect immune activation by recombinant Galleria mellonella apolipophorin III. Biochim. Biophys. Acta 1433(1–2), 16–26 (1999).
- 36. Immune activation of apolipophorin-III and its distribution in hemocyte from Hyphantria cunea. Insect Biochem. Mol. Biol. 34(10), 1011–1023 (2004).
- 37. . Synergistic action of Galleria mellonella apolipophorin III and lysozyme against Gram-negative bacteria. Biochim. Biophys. Acta 1828(6), 1449–1456 (2013).
- 38. . Involvement of apolipophorin III in antibacterial defense of Galleria mellonella larvae. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 158(1), 90–98 (2011).
- 39. . The effect of Galleria mellonella apolipophorin III on yeasts and filamentous fungi. J. Insect Physiol. 58(1), 164–177 (2012).
- 40. . Galleria mellonella apolipophorin III – an apolipoprotein with anti-Legionella pneumophila activity. Biochim. Biophys. Acta 1838(10), 2689–2697 (2014). • Reports on anti-Legionella pneumophila activity of insect apolipophorin III and possible mechanism responsible for this activity.
- 41. . Anti-Legionella dumoffii activity of Galleria mellonella defensin and apolipophorin III. Int. J. Mol. Sci. 13(12), 17048–17064 (2012).
- 42. The lipid composition of Legionella dumoffii membrane modulates the interaction with Galleria mellonella apolipophorin III. Biochim. Biophys. Acta 1861(7), 617–629 (2016).
- 43. . Rapid extraction of bacterial genomic DNA with guanidium thiocyanate. Lett. Appl. Microbiol. 8(4), 151–156 (1989).
- 44. 16S ribosomal DNA amplification for phylogenetic study. J. Bacteriol. 173(2), 697–703 (1991).
- 45. . Molecular Detection of Human Bacterial Pathogens. CRC Press, Boca Raton, FL, USA (2011).
- 46. . CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22, 4673–4680 (1994).
- 47. . GeneDoc. Pittsburgh Supercomputing Center, Pittsburgh, PA, USA (1997).
- 48. . MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol. Biol. Evol. 30(12), 2725–2729 (2013).
- 49. . PHYLIP. Phylogeny Inference Package, version 3.5c., University of Washington, Seattle, WA, USA (1993).
- 50. . A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J. Mol. Evol. 16(2), 111–120 (1980).
- 51. . TreeView: an application to display phylogenetic trees on personal computers. Comput. Appl. Biosci. 12(4), 357–358 (1996).
- 52. . Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal. Biochem. 166, 368–379 (1987).
- 53. . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254 (1976).
- 54. . Infrared spectroscopy of proteins and peptides in lipid bilayers. Quart. Rev. Biophys. 30, 365–429 (1997).
- 55. . Native valve Staphylococcus capitis infective endocarditis: a mini review. Infection 48(1), 3–5 (2020).
- 56. . Coagulase-negative staphylococci pathogenomics. Int. J. Mol. Sci. 20(5), 1215 (2019).
- 57. . Coagulase-negative Staphylococcus skin and soft tissue infections. Am. J. Clin. Dermatol. 19(5), 671–677 (2018). • Coagulase-negative staphylococcal organisms should not always be considered as contaminants or normal flora, but rather as causative pathogens of human skin and soft tissue.
- 58. . Staphylococcal biofilms in atopic dermatitis. Curr. Allergy Asthma Rep. 17(12), 81 (2017). • Staphylococcal colonization of the skin impacts skin barrier function and plays multiple important roles in atopic dermatitis pathogenesis.
- 59. Antibiotic resistance of commensal Staphylococcus aureus and coagulase-negative Staphylococci in an international cohort of surgeons: a prospective point-prevalence study. PLoS ONE 11(2), e0148437 (2016).
- 60. . Comparative analysis of Staphylococcus epidermidis strains utilizing quantitative and cell surface shaving proteomics. J. Proteomics 130, 190–199 (2016).
- 61. . LL-37 fragments have antimicrobial activity against Staphylococcus epidermidis biofilms and wound healing potential in HaCaT cell line. J. Pep. Sci. 24(7), e3080 (2018).
- 62. . Mechanisms of resistance to antimicrobial peptides in staphylococci. Biochim. Biophys. Acta 1848(11), 3055–3061 (2015).
- 63. . The human anionic antimicrobial peptide dermcidin induces proteolytic defence mechanisms in staphylococci. Mol. Microbiol. 63(2), 497–506 (2007).
- 64. . Staphylococcus epidermidis – the ‘accidental’ pathogen. Nat. Rev. Microbiol. 7(8), 555–567 (2009). • Describes the molecular basis of the commensal and infectious lifestyles of Staphylococcus epidermidis.