1887

Abstract

This study detailed the responses of larvae to disseminated infection caused by co-infection with and . Doses of (1×10 larva) and (1×10 larva) were non-lethal in mono-infection but when combined significantly (<0.05) reduced larval survival at 24, 48 and 72 h relative to larvae receiving (2×10 larva) alone. Co-infected larvae displayed a significantly higher density of larva compared to larvae infected solely with . Co-infection resulted in dissemination throughout the host and the appearance of large nodules. Co-infection of larvae with and (2×10 larva) resulted in an increase in the density of circulating haemocytes compared to that in larvae infected with only . Proteomic analysis of co-infected larval haemolymph revealed increased abundance of proteins associated with immune responses to bacterial and fungal infection such as cecropin-A (+45.4-fold), recognition proteins [e.g. peptidoglycan-recognition protein LB (+14-fold)] and proteins associated with nodule formation [e.g. Hdd11 (+33.3-fold)]. A range of proteins were also decreased in abundance following co-infection, including apolipophorin (−62.4-fold), alpha-esterase 45 (−7.7-fold) and serine proteinase (−6.2-fold). Co-infection of larvae resulted in enhanced proliferation of compared to mono-infection and an immune response showing many similarities to the innate immune response of mammals to infection. The utility of larvae for studying polymicrobial infection is highlighted.

Funding
This study was supported by the:
  • Science Foundation Ireland (Award 12/RI/2346 (3).)
    • Principle Award Recipient: Not Applicable
  • Science Foundation Ireland (Award 12/RC/2275_P2.)
    • Principle Award Recipient: Kevin A. Kavanagh
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000892
2020-02-18
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/166/4/375.html?itemId=/content/journal/micro/10.1099/mic.0.000892&mimeType=html&fmt=ahah

References

  1. Shirtliff ME, Peters BM, Jabra-Rizk MA. Cross-kingdom interactions: Candida albicans and bacteria. FEMS Microbiol Lett 2009; 299:1–8 [View Article][PubMed]
    [Google Scholar]
  2. Reece E, Segurado R, Jackson A, Mcclean S, Renwick J et al. Co-colonisation with Aspergillus fumigatus and Pseudomonas aeruginosa is associated with poorer health in cystic fibrosis patients : an Irish registry analysis. BMC Pulm Med 20171–8
    [Google Scholar]
  3. Klotz SA, Chasin BS, Powell B, Gaur NK, Lipke PN. Polymicrobial bloodstream infections involving Candida species: analysis of patients and review of the literature. Diagn Microbiol Infect Dis 2007; 59:401–406 [View Article]
    [Google Scholar]
  4. Reno J, Doshi S, Tunali AK, Stein B, Farley MM et al. Epidemiology of methicillin-resistant Staphylococcus aureus bloodstream coinfection among adults with candidemia in atlanta, GA, 2008–2012. Infect Control Hosp Epidemiol 2015; 36:1298–1304 [View Article]
    [Google Scholar]
  5. O'Donnell LE, Millhouse E, Sherry L, Kean R, Malcolm J et al. Polymicrobial Candida biofilms: friends and foe in the oral cavity. FEMS Yeast Res 2015; 15:fov077 [View Article]
    [Google Scholar]
  6. Bertolini M, Ranjan A, Thompson A, Diaz PI, Sobue T et al. Candida albicans induces mucosal bacterial dysbiosis that promotes invasive infection. PLoS Pathog 2019; 15:e1007717 [View Article]
    [Google Scholar]
  7. Diaz PI, Strausbaugh LD, Dongari-Bagtzoglou A. Fungal-bacterial interactions and their relevance to oral health: linking the clinic and the bench. Front Cell Infect Microbiol 2014; 29:101
    [Google Scholar]
  8. Schlecht LM, Peters BM, Krom BP, Freiberg JA, Hänsch GM et al. Systemic Staphylococcus aureus infection mediated by Candida albicans hyphal invasion of mucosal tissue. Microbiology 2015; 161:168–181 [View Article]
    [Google Scholar]
  9. Kong EF, Tsui C, Kucharíková S, Van Dijck P, Jabra-Rizk MA. Modulation of Staphylococcus aureus response to antimicrobials by the Candida albicans quorum sensing molecule farnesol. Antimicrob Agents Chemother 2017; 61: [View Article]
    [Google Scholar]
  10. Allison DL, Scheres N, Willems HME, Bode CS, Krom BP et al. The host immune system facilitates disseminated Staphylococcus aureus disease due to phagocytic attraction to Candida albicans during coinfection: a case of bait and switch. Infect Immun 2019; 87: [View Article]
    [Google Scholar]
  11. Todd OA, Peters BM. Candida albicans and Staphylococcus aureus pathogenicity and polymicrobial interactions: lessons beyond koch’s postulates. Journal of Fungi 2019; 5:81 [View Article]
    [Google Scholar]
  12. Kong EF, Tsui C, Kucharíková S, Andes D, Van Dijck P et al. Commensal Protection of Staphylococcus aureus against Antimicrobials by Candida albicans Biofilm Matrix. MBio 2016; 7: [View Article]
    [Google Scholar]
  13. Harriott MM, Noverr MC. Candida albicans and Staphylococcus aureus form polymicrobial biofilms: effects on antimicrobial resistance. Antimicrob Agents Chemother 2009; 53:3914–3922 [View Article]
    [Google Scholar]
  14. Kean R, Rajendran R, Haggarty J, Townsend EM, Short B et al. Candida albicans mycofilms support Staphylococcus aureus colonization and enhances miconazole resistance in dual-species interactions. Front Microbiol 2017; 8: [View Article]
    [Google Scholar]
  15. de Carvalho Dias K, Barbugli PA, de Patto F, Lordello VB, de Aquino Penteado L et al. Soluble factors from biofilm of Candida albicans and Staphylococcus aureus promote cell death and inflammatory response. BMC Microbiol 2017; 17: [View Article]
    [Google Scholar]
  16. Krüger W, Vielreicher S, Kapitan M, Jacobsen ID, Niemiec MJ. Fungal-Bacterial interactions in health and disease. Pathogens 2019; 8:70 [View Article]
    [Google Scholar]
  17. Todd OA, Fidel PL, Harro JM, Hilliard JJ, Tkaczyk C et al. Candida albicans augments Staphylococcus aureus virulence by engaging the Staphylococcal agr quorum sensing system. MBio 2019; 10: [View Article]
    [Google Scholar]
  18. Carolus H, Van Dyck K, Van Dijck P, albicans C. Candida albicans and Staphylococcus species: a threatening Twosome. Front Microbiol 2019; 10: [View Article]
    [Google Scholar]
  19. Ikeh MAC, Fidel PL, Noverr MC. Identification of specific components of the eicosanoid biosynthetic and signaling pathway involved in pathological inflammation during intra-abdominal infection with Candida albicans and Staphylococcus aureus . Infect Immun 2018; 86: [View Article]
    [Google Scholar]
  20. Peters BM, Jabra-Rizk MA, Scheper MA, Leid JG, Costerton JW et al. Microbial interactions and differential protein expression in Staphylococcus aureus–Candida albicans dual-species biofilms. FEMS Immunol Med Microbiol 2010; 59:493–503 [View Article]
    [Google Scholar]
  21. Kasper L, König A, Koenig P-A, Gresnigt MS, Westman J et al. The fungal peptide toxin Candidalysin activates the NLRP3 inflammasome and causes cytolysis in mononuclear phagocytes. Nat Commun 2018; 9: [View Article]
    [Google Scholar]
  22. Mayer FL, Wilson D, Hube B. Candida albicans pathogenicity mechanisms. Virulence 2013; 4:119–128 [View Article]
    [Google Scholar]
  23. Lacey K, Geoghegan J, McLoughlin R. The role of Staphylococcus aureus virulence factors in skin infection and their potential as vaccine antigens. Pathogens 2016; 5:22 [View Article]
    [Google Scholar]
  24. Shinji H, Yosizawa Y, Tajima A, Iwase T, Sugimoto S et al. Role of Fibronectin-Binding Proteins A and B in In Vitro Cellular Infections and In Vivo Septic Infections by Staphylococcus aureus . Infect Immun 2011; 79:2215–2223 [View Article]
    [Google Scholar]
  25. Bhakdi S, Tranum-Jensen J. Alpha-Toxin of Staphylococcus aureus . Microbiol Rev 1991; 55:733–751 [View Article]
    [Google Scholar]
  26. Burman JD, Leung E, Atkins KL, O’Seaghdha MN, Lango L et al. Interaction of human complement with Sbi, a staphylococcal immunoglobulin-binding protein: indications of a novel mechanism of complement evasion by Staphylococcus aureus . J Biol Chem 2008; 283:17579–17593
    [Google Scholar]
  27. Jongerius I, Köhl Jörg, Pandey MK, Ruyken M, van Kessel KPM et al. Staphylococcal complement evasion by various convertase-blocking molecules. J Exp Med 2007; 204:2461–2471 [View Article]
    [Google Scholar]
  28. Jardeleza C, Jones D, Baker L, Miljkovic D, Boase S et al. Gene expression differences in nitric oxide and reactive oxygen species regulation point to an altered innate immune response in chronic rhinosinusitis. Int Forum Allergy Rhinol 2013; 3:193–198 [View Article]
    [Google Scholar]
  29. Tsai CJ-Y, Loh JMS, Proft T, Jia C, -Yun T, Mei J, Loh S. Galleria mellonella infection models for the study of bacterial diseases and for antimicrobial drug testing. Virulence 2016; 7:214–229 [View Article][PubMed]
    [Google Scholar]
  30. Senior NJ, Bagnall MC, Champion OL, Reynolds SE, La Ragione RM et al. Galleria mellonella as an infection model for Campylobacter jejuni virulence. J Med Microbiol 2011; 60:661–669 [View Article]
    [Google Scholar]
  31. Borman AM, Szekely A, Johnson EM. Comparative Pathogenicity of United Kingdom Isolates of the Emerging Pathogen Candida auris and Other Key Pathogenic Candida Species. mSphere 2016; 1:e00189–16 [View Article]
    [Google Scholar]
  32. Kavanagh K, Sheehan G. The use of Galleria mellonella larvae to identify novel antimicrobial agents against fungal species of medical interest. JoF 2018; 4:113 [View Article]
    [Google Scholar]
  33. Mylonakis E, Casadevall A, Ausubel FM. Exploiting amoeboid and non-vertebrate animal model systems to study the virulence of human pathogenic fungi. PLoS Pathog 2007; 3:e101 [View Article]
    [Google Scholar]
  34. Sheehan G, Dixon A, Kavanagh K. Utilization of Galleria mellonella larvae to characterize the development of Staphylococcus aureus infection. Microbiology 2019; 165:863–875 [View Article]
    [Google Scholar]
  35. Evans BA, Rozen DE. A Streptococcus pneumoniae infection model in larvae of the wax moth Galleria mellonella. Eur J Clin Microbiol Infect Dis 2012; 31:2653–2660 [View Article]
    [Google Scholar]
  36. La Rosa SL, Casey PG, Hill C, Diep DB, Nes IF et al. In Vivo assessment of growth and virulence gene expression during commensal and pathogenic lifestyles of luxABCDE -Tagged Enterococcus faecalis strains in murine gastrointestinal and intravenous infection models. Appl Environ Microbiol 2013; 79:3986–3997 [View Article]
    [Google Scholar]
  37. Leanti La Rosa S, Diep DB, Nes IF, Brede DA. Construction and Application of a luxABCDE reporter system for real-time monitoring of enterococcus faecalis gene expression and growth. Appl Environ Microbiol 2012; 78:7003–7011 [View Article]
    [Google Scholar]
  38. Sheehan G, Kavanagh K. Analysis of the early cellular and humoral responses of Galleria mellonella larvae to infection by Candida albicans . Virulence 2018; 9:163–172 [View Article]
    [Google Scholar]
  39. Sheehan G, Kavanagh K. Proteomic analysis of the responses of Candida albicans during infection of Galleria mellonella larvae. JoF 2019; 5:7 [View Article]
    [Google Scholar]
  40. Sheehan G, Clarke G, Kavanagh K. Characterisation of the cellular and proteomic response of Galleria mellonella larvae to the development of invasive aspergillosis. BMC Microbiol 2018; 18: [View Article]
    [Google Scholar]
  41. London R, Orozco BS, Mylonakis E. The pursuit of cryptococcal pathogenesis: heterologous hosts and the study of cryptococcal host–pathogen interactions. FEMS Yeast Res 2006; 6:567–573 [View Article]
    [Google Scholar]
  42. Silva LN, Da Hora GCA, Soares TA, Bojer MS, Ingmer H et al. Myricetin protects Galleria mellonella against Staphylococcus aureus infection and inhibits multiple virulence factors. Sci Rep 2017; 7: [View Article]
    [Google Scholar]
  43. Cox Jürgen, Neuhauser N, Michalski A, Scheltema RA, Olsen JV et al. Andromeda: a peptide search engine integrated into the MaxQuant environment. J Proteome Res 2011; 10:1794–1805
    [Google Scholar]
  44. Vogel H, Altincicek B, Glöckner G, Vilcinskas A. A comprehensive transcriptome and immune-gene repertoire of the lepidopteran model host Galleria mellonella. BMC Genomics 2011; 12:308 [View Article]
    [Google Scholar]
  45. Côté RG, Griss J, Dianes JA, Wang R, Wright JC et al. The proteomics identification (pride) converter 2 framework: an improved suite of tools to facilitate data submission to the pride database and the ProteomeXchange Consortium. Mol Cell Proteomics 2012; 11:1682–1689 [View Article]
    [Google Scholar]
  46. Maguire R, Kunc M, Hyrsl P, Kavanagh K. Caffeine administration alters the behaviour and development of Galleria mellonella larvae. Neurotoxicol Teratol 2017; 64:37–44 [View Article]
    [Google Scholar]
  47. Gu W, Yu Q, Yu C, Sun S. In vivo activity of fluconazole/tetracycline combinations in Galleria mellonella with resistant Candida albicans infection. J Glob Antimicrob Resist 2018; 13:74–80 [View Article][PubMed]
    [Google Scholar]
  48. Mylonakis E, Moreno R, El Khoury JB, Idnurm A, Heitman J et al. Galleria mellonella as a model system to study Cryptococcus neoformans pathogenesis. Infect Immun 2005; 73:3842–3850 [View Article]
    [Google Scholar]
  49. Lim W, Melse Y, Konings M, Phat Duong H, Eadie K et al. Addressing the most neglected diseases through an open research model: the discovery of fenarimols as novel drug candidates for eumycetoma. PLoS Negl Trop Dis 2018; 12:e0006437 [View Article]
    [Google Scholar]
  50. Peters BM, Noverr MC. Candida albicans-staphylococcus aureus polymicrobial peritonitis modulates host innate immunity. Infect Immun 2013; 81:2178–2189 [View Article]
    [Google Scholar]
  51. Ratcliffe NA. Invertebrate immunity — a primer for the non-specialist. Immunol Lett 1985; 10:253–270 [View Article]
    [Google Scholar]
  52. Yi HY, Deng XJ, Yang WY, Zhou CZ, Cao Y et al. Gloverins of the silkworm Bombyx mori: structural and binding properties and activities. Insect Biochem Mol Biol 2013; 43:612–625 [View Article][PubMed]
    [Google Scholar]
  53. XX X, Zhong X, HY Y, XQ Y. Manduca sexta gloverin binds microbial components and is active against bacteria and fungi. Dev Comp Immunol 2012; 38:275–284
    [Google Scholar]
  54. Sheehan G, Garvey A, Croke M, Kavanagh K. Innate humoral immune defences in mammals and insects: The same, with differences ?. Virulence 2018; 9:1625–1639 [View Article]
    [Google Scholar]
  55. Yun J, Lee DG. Cecropin A-induced apoptosis is regulated by ion balance and glutathione antioxidant system in Candida albicans . IUBMB Life 2016; 68:652–662 [View Article]
    [Google Scholar]
  56. Lee E, Shin A, Kim Y. Anti-Inflammatory activities of cecropin A and its mechanism of action. Arch Insect Biochem Physiol 2015; 88:31–44 [View Article]
    [Google Scholar]
  57. Lee E, Jeong K-W, Lee J, Shin A, Kim J-K et al. Structure-Activity relationships of cecropin-like peptides and their interactions with phospholipid membrane. BMB Rep 2013; 46:282–287 [View Article]
    [Google Scholar]
  58. Hara S, Moricin YM. A novel type of antibacterial peptide isolated from the silkworm, Bombyx mori. J Biol Chem 1995; 270:29923–29927
    [Google Scholar]
  59. Brown SE, Howard A, Kasprzak AB, Gordon KH, East PD. The discovery and analysis of a diverged family of novel antifungal moricin-like peptides in the wax moth Galleria mellonella. Insect Biochem Mol Biol 2008; 38:201–212 [View Article]
    [Google Scholar]
  60. Shin SW, Park SS, Park DS, Kim MG, Kim SC, Woon Shin S, Park D-S, Gwang Kim M et al. Isolation and characterization of immune-related genes from the fall webworm, Hyphantria cunea, using PCR-based differential display and subtractive cloning. Insect Biochem Mol Biol 1998; 28:827–837 [View Article][PubMed]
    [Google Scholar]
  61. Sarauer BL, Gillott C, Hegedus D. Characterization of an intestinal mucin from the peritrophic matrix of the diamondback moth, Plutella xylostella. Insect Mol Biol 2003; 12:333–343 [View Article]
    [Google Scholar]
  62. Gandhe AS, John SH, Nagaraju J, Noduler NJ. Noduler, a novel immune up-regulated protein mediates nodulation response in insects. J Immunol 2007; 179:6943–6951 [View Article]
    [Google Scholar]
  63. Niere M, Meißlitzer C, Dettloff M, Weise C, Ziegler M et al. Insect immune activation by recombinant Galleria mellonella apolipophorin III. Biochim Biophys Acta 1999; 1433:16–26 [View Article]
    [Google Scholar]
  64. Niere M, Dettloff M, Maier T, Ziegler M, Wiesner A. Insect immune activation by apolipophorin III is correlated with the lipid-binding properties of this protein. Biochemistry 2001; 40:11502–11508 [View Article][PubMed]
    [Google Scholar]
  65. Zdybicka-Barabas A, Cytryñska M. Apolipophorins and insects immune response. ISJ - Invertebr Surviv J 2013; 10:58–68
    [Google Scholar]
  66. Park SY, Kim CH, Jeong WH, Lee JH, Seo SJ et al. Effects of two hemolymph proteins on humoral defense reactions in the wax moth, Galleria mellonella. Dev Comp Immunol 2005
    [Google Scholar]
  67. Zdybicka-Barabas A, Cytryńska M. Involvement of apolipophorin III in antibacterial defense of Galleria mellonella larvae. Comp Biochem Physiol B Biochem Mol Biol 2011; 158:90–98 [View Article]
    [Google Scholar]
  68. Zdybicka-Barabas A, Mak P, Jakubowicz T, Cytryńska M. Lysozyme and defense peptides as suppressors of phenoloxidase activity in Galleria mellonella . Arch Insect Biochem Physiol 2014; 87:1–12 [View Article][PubMed]
    [Google Scholar]
  69. Wojda I. Immunity of the greater wax moth Galleria mellonella . Insect Sci 2017; 24:342–357 [View Article]
    [Google Scholar]
  70. Sun JN, Solis NV, Phan QT, Bajwa JS, Kashleva H et al. Host cell invasion and virulence mediated by Candida albicans Ssa1. PLoS Pathog 2010; 6:e1001181 [View Article]
    [Google Scholar]
  71. Cho T, Toyoda M, Sudoh M, Nakashima Y, Calderone RA et al. Isolation and sequencing of the Candida albicans MSI3, a putative novel member of the hsp70 family. Yeast 2003; 20:149–156 [View Article]
    [Google Scholar]
  72. Li XS, Reddy MS, Baev D, Edgerton M. Candida albicans Ssa1/2p is the cell envelope binding protein for human salivary histatin 5. J Biol Chem 2003; 278:28553–28561 [View Article][PubMed]
    [Google Scholar]
  73. Nagao J-ichi, Cho T, Uno J, Ueno K, Imayoshi R et al. Candida albicans Msi3p, a homolog of the Saccharomyces cerevisiae Sse1p of the hsp70 family, is involved in cell growth and fluconazole tolerance. FEMS Yeast Res 2012; 12:728–737
    [Google Scholar]
  74. Urban C, Xiong X, Sohn K, Schröppel K, Brunner H et al. The moonlighting protein Tsa1p is implicated in oxidative stress response and in cell wall biogenesis in Candida albicans . Mol Microbiol 2005; 57:1318–1341 [View Article]
    [Google Scholar]
  75. Smagur J, Guzik K, Bzowska M, Kuzak M, Zarebski M et al. Staphylococcal cysteine protease staphopain B (SspB) induces rapid engulfment of human neutrophils and monocytes by macrophages. Biol Chem 2009; 390:361–371 [View Article]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000892
Loading
/content/journal/micro/10.1099/mic.0.000892
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error