Evaluation of the antagonistic effect of Pseudomonas aeruginosa toxins on azole antifungal resistance in Candida albicans species isolated from clinical samples in Iran
-
Masoumeh Sadat Hosseini
,
Masoumeh Navidinia
,
Sima Sadat Seyedjavadi
,
Mehdi Goudarzi
,
Helia Rasouli
,
Amir Mohsen Mahdavian
,
Elina Rahimi Zamani
Abstract
Background and Objectives: The azole antifungals are the most frequent class used to treat Candida infections. It is essential to elucidate the potential of natural compounds as an alternative in eliminating Candida albicans (C. albicans). Therefore, in the present study, the antagonistic effect of Pseudomonas aeruginosa toxins on azole antifungal resistance in C. albicans species was investigated.
Materials and Methods: In this study, 28 C. albicans species with azole antifungal resistance were obtained from patients at Shohadaye Tajrish Hospital. The effect of toxins, such as phenazine, pyocyanin, pyoverdine, and fluorescein, was examined on C. albicans species. The antifungal activity of these toxins against C. albicans spp. was determined using methods such as minimal inhibitory concentration (MIC90), radial diffusion assay (RDA), and detection of reactive oxygen species (ROS).
Results: The prevalence of C. albicans strains in urinary catheters, surgical wounds, respiratory tracts, blood, and standard strains was 46.3%, 21.4%, 25%, 7.14%, and 3.57%, respectively. The MIC values were reported as 32 µg/ml for phenazine, and 128 µg/ml for pyoverdine, pyocyanin, and fluorescein. The results showed that phenazine exhibited higher inhibitory effects against C. albicans isolated from clinical samples compared to the other toxins. After exposure to phenazines (20 µg/ml), 65-70% of yeast cells of C. albicans spp. showed rhodamine 123 fluorescence, indicating high intracellular reactive oxygen species (ROS) production.
Conclusion: The antifungal effect of different toxins in C. albicans spp. may be due to ROS-mediated apoptotic death. The results suggest that phenazine has high potential in controlling C. albicans. This natural compounds are a potential alternative for eliminating this yeast.
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10. Zarrinfar H, Kord Z, Fata A. High incidence of azole resistance among C. albicans and C. glabrata isolates in Northeastern Iran. Curr Med Mycol 2021; 7: 18-21.
11. Kermani F, Taghizadeh-Armaki M, Hosseini SA, Amirrajab N, Javidnia J, Fami Zaghrami M, et al. Antifungal resistance of clinical Candida albicans isolates in Iran: A systematic review and Meta-Analysis. Iran J Public Health 2023; 52: 290-305.
12. 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; 4: 101.
13. Shirtliff ME, Peters BM, Jabra-Rizk MA. Cross-kingdom interactions: C. albicans and bacteria. FEMS Microbiol Lett 2009; 299: 1-8.
14. Dhamgaye S, Qu Y, Peleg AY. Polymicrobial infections involving clinically relevant Gram-negative bacteria and fungi. Cell Microbiol 2016; 18: 1716-1722.
15. Singh A, Verma R, Murari A, Agrawal A .Oral candidiasis: an overview. J Oral Maxillofac Pathol 2014; 18(Suppl 1): S81-S85.
16. Ahmad S, Khan Z. Invasive candidiasis: a review of nonculture-based laboratory diagnostic methods. Indian J Med Microbiol 2012; 30: 264-269.
17. Moya-Salazar J, Rojas R. Comparative study for identification of C. albicans with germ tube test in human serum and plasma. Clin Microbiol Infect Dis 2018; 3: 1-4.
18. Shin JH, Nolte FS, Holloway BP, Morrison CJ. Rapid identification of up to three Candida species in a single reaction tube by a 5′ exonuclease assay using fluorescent DNA probes. J Clin Microbiol 1999; 37: 165-170.
19. Latouche GN, Daniel HM, Lee OC, Mitchell TG, Sorrell TC, Meyer W. Comparison of use of phenotypic and genotypic characteristics for identification of species of the anamorph genus Candida and related teleomorph yeast species. J Clin Microbiol 1997; 35: 3171-3180.
20. Musinguzi BJ, Sande O, Mboowa G, Baguma A, Itabangi H, Achan B. Laboratory diagnosis of Candidiasis. Candida and Candidiasis. IntechOpen; 2023. doi:10.5772/intechopen.106359.
21. Díez A, Carrano G, Bregón-Villahoz M, Cuétara MS, García-Ruiz JC, Fernandez-de-Larrinoa I, et al. Biomarkers for the diagnosis of invasive candidiasis in immunocompetent and immunocompromised patients. Diagn Microbiol Infect Dis 2021; 101: 115509.
22. Roberts GD, Wang HS, Hollick GE. Evaluation of the API 20 C microtube system for the identification of clinically important yeasts. J Clin Microbiol 1976; 3: 302-305.
23. Takemura H, Kaku M, Kohno S, Hirakata Y, Tanaka H, Yoshida R, et al. Evaluation of susceptibility of gram-positive and -negative bacteria to human defensins by using radial diffusion assay. Antimicrob Agents Chemother 1996; 40: 2280-2284.
24. Liang X, Yan J, Lu Y, Liu S, Chai X. The antimicrobial peptide melectin shows both antimicrobial and antitumor activity via membrane interference and DNA binding. Drug Des Devel Ther 2021; 15: 1261-1273.
25. Costa CR, Jesuíno RS, de Aquino Lemos J, de Fátima Lisboa Fernandes O, Hasimoto e Souza LK, Passos XS, et al. Effects of antifungal agents in sap activity of Candida albicans isolates. Mycopathologia 2010; 169: 91-98.
26. CLSI (2022). Performance Standards for Antifungal Susceptibility Testing of Yeasts. 3rd ed. CLSI supplement M27M44S. Replaces M60-Ed2. Clinical and Laboratory Standards Institute. https://clsi.org/media/osthhxax/m27m44sed3e_sample.pdf
27. Su L, Zhang J, Gomez H, Kellum JA, Peng Z. Mitochondria ROS and mitophagy in acute kidney injury. Autophagy 2023; 19: 401-414.
28. Grainha T, Jorge P, Alves D, Lopes SP, Pereira MO. Unraveling Pseudomonas aeruginosa and Candida albicans communication in coinfection scenarios: Insights through network analysis. Front Cell Infect Microbiol 2020; 10: 550505.
29. Hogan DA, Kolter R. Pseudomonas-Candida interactions: an ecological role for virulence factors. Science 2002; 296: 2229-2232.
30. Lindsay AK, Deveau A, Piispanen AE, Hogan DA. Farnesol and cyclic AMP signaling effects on the hypha-to-yeast transition in Candida albicans. Eukaryot Cell 2012; 11: 1219-1225.
31. Grainha T, Jorge P, Alves D, Lopes SP, Pereira MO. Unraveling Pseudomonas aeruginosa and Candida albicans Communication in coinfection scenarios: Insights through network analysis. Front Cell Infect Microbiol 2020; 10: 550505.
32. Rueda C, Puig-Asensio M, Guinea J, Almirante B, Cuenca-Estrella M, Zaragoza O, et al. Evaluation of the possible influence of trailing and paradoxical effects on the clinical outcome of patients with candidemia. Clin Microbiol Infect 2017; 23: 49.e1-49.e8.
33. Marr KA, Rustad TR, Rex JH, White TC. The trailing end point phenotype in antifungal susceptibility testing is pH dependent. Antimicrob Agents Chemother 1999; 43: 1383-1386.
34. Marcos-Zambrano LJ, Escribano P, Sánchez-Carrillo C, Bouza E, Guinea J. Scope and frequency of fluconazole trailing assessed using Eucast in invasive Candida spp. isolates. Med Mycol 2016; 54: 733-739.
35. Lee MK, Williams LE, Warnock DW, Arthington-Skaggs BA. Drug resistance genes and trailing growth in Candida albicans isolates. J Antimicrob Chemother 2004; 53: 217-224.
36. Arthington-Skaggs BA, Lee-Yang W, Ciblak MA, Frade JP, Brandt ME, Hajjeh RA, et al. Comparison of visual and spectrophotometric methods of broth microdilution MIC end point determination and evaluation of a sterol quantitation method for in vitro susceptibility testing of fluconazole and itraconazole against trailing and nontrailing Candida isolates. Antimicrob Agents Chemother 2002; 46: 2477–2481.
37. Lee MK, Kim HR, Kang JO, Kim MN, Kim EC, Kim JS, et al. Susceptibility and trailing growth of Candida albicans to fluconazole: Results of a korean multicentre study. Mycoses 2007; 50: 148-149.
38. Arthington-Skaggs BA, Warnock DW, Morrison CJ. Quantitation of Candida albicans ergosterol content improves the correlation between in vitro antifungal susceptibility test results and in vivo outcome after fluconazole treatment in a murine model of invasive candidiasis. Antimicrob Agents Chemother 2000; 44: 2081-2085.
39. Odabasi Z, Paetznick VL, Rodriguez JR, Chen E, Rex JH, Leitz GJ, et al. Lack of correlation of 24- vs. 48-h itraconazole minimum inhibitory concentrations with microbiological and survival outcomes in a Guinea pig model of disseminated candidiasis. Mycoses 2010; 53: 438-442.
40. Binder U, Aigner M, Risslegger B, Hörtnagl C, Lass-Flörl C, Lackner M. Minimal Inhibitory Concentration (MIC)-Phenomena in Candida albicans and their impact on the diagnosis of antifungal resistance. J Fungi (Basel) 2019; 5: 83.
41. Tupe SG, Kulkarni RR, Shirazi F, Sant DG, Joshi SP, Deshpande MV. Possible mechanism of antifungal phenazine-1-carboxamide from Pseudomonas sp. against dimorphic fungi Benjaminiella poitrasii and human pathogen Candida albicans. J Appl Microbiol 2015; 118: 39-48.
42. Alam F, Blackburn SA, Davis J, Massar K, Correia J, Tsai HJ, et al. Pseudomonas aeruginosa increases the susceptibility of Candida albicans to amphotericin B in dual-species biofilms. J Antimicrob Chemother 2023; 78: 2228-2241.
2. Horn DL, Neofytos D, Anaissie EJ, Fishman JA, Steinbach WJ, Olyaei AJ. Epidemiology and outcomes of candidemia in 2019 patients: data from the prospective antifungal therapy alliance registry. Clin Infect Dis 2009; 48: 1695-1703.
3. Kahl LJ, Stremmel N, Esparza-Mora MA, Wheatley RM, MacLean RC, Rasler M. Interkingdom interactions between Pseudomonas aeruginosa and Candida albicans affect clinical outcomes and antimicrobial responses. Curr Opin Microbiol 2023; 75: 102368.
4. Hattab S, Dagher AM, Wheelera RT. Pseudomonas Synergizes with Fluconazole against Candida during Treatment of Polymicrobial infection. Infect Immun 2022; 90(4): e0062621.
5. Chaudhary PM, Chavan SR, Shirazi F, Razdan M, Nimkar P, Maybhate SP, et al. Exploration of click reaction for the synthesis of modified nucleosides as chitin synthase inhibitors. Bioorg Med Chem 2009; 17: 2433-2440.
6. Agarwal H, Bajpai S, Mishra A, Kohli I, Varma A, Fouillaud M, et al. Bacterial Pigments and their multifaceted roles in contemporary Biotechnology and Pharmacological applications. Microorganisms 2023; 11: 614.
7. Houshaymi B, Awada R, Kedees M, Soayfane Z. Pyocyanin, a Metabolite of Pseudomonas aeruginosa, Exhibits Antifungal drug activity through Inhibition of a Pleiotropic drug resistance Subfamily FgABC3. Drug Res (Stuttg) 2019; 69: 658-664.
8. Agarwal H, Bajpai S, Mishra A, Kohli I, Varma A, Fouillaud M, et al. Bacterial Pigments and their Multifaceted roles in contemporary Biotechnology and Pharmacological applications. Microorganisms 2023; 11: 614.
9. Mirsalehian A, Feizabadi M, Nakhjavani FA, Jabalameli F, Goli H, Kalantari N, et al. Detection of VEB-1, OXA-10 and PER-1 genotypes in extended-spectrum beta-lactamase-producing Pseudomonas aeruginosa strains isolated from burn patients. Burns 2010; 36: 70-74.
10. Zarrinfar H, Kord Z, Fata A. High incidence of azole resistance among C. albicans and C. glabrata isolates in Northeastern Iran. Curr Med Mycol 2021; 7: 18-21.
11. Kermani F, Taghizadeh-Armaki M, Hosseini SA, Amirrajab N, Javidnia J, Fami Zaghrami M, et al. Antifungal resistance of clinical Candida albicans isolates in Iran: A systematic review and Meta-Analysis. Iran J Public Health 2023; 52: 290-305.
12. 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; 4: 101.
13. Shirtliff ME, Peters BM, Jabra-Rizk MA. Cross-kingdom interactions: C. albicans and bacteria. FEMS Microbiol Lett 2009; 299: 1-8.
14. Dhamgaye S, Qu Y, Peleg AY. Polymicrobial infections involving clinically relevant Gram-negative bacteria and fungi. Cell Microbiol 2016; 18: 1716-1722.
15. Singh A, Verma R, Murari A, Agrawal A .Oral candidiasis: an overview. J Oral Maxillofac Pathol 2014; 18(Suppl 1): S81-S85.
16. Ahmad S, Khan Z. Invasive candidiasis: a review of nonculture-based laboratory diagnostic methods. Indian J Med Microbiol 2012; 30: 264-269.
17. Moya-Salazar J, Rojas R. Comparative study for identification of C. albicans with germ tube test in human serum and plasma. Clin Microbiol Infect Dis 2018; 3: 1-4.
18. Shin JH, Nolte FS, Holloway BP, Morrison CJ. Rapid identification of up to three Candida species in a single reaction tube by a 5′ exonuclease assay using fluorescent DNA probes. J Clin Microbiol 1999; 37: 165-170.
19. Latouche GN, Daniel HM, Lee OC, Mitchell TG, Sorrell TC, Meyer W. Comparison of use of phenotypic and genotypic characteristics for identification of species of the anamorph genus Candida and related teleomorph yeast species. J Clin Microbiol 1997; 35: 3171-3180.
20. Musinguzi BJ, Sande O, Mboowa G, Baguma A, Itabangi H, Achan B. Laboratory diagnosis of Candidiasis. Candida and Candidiasis. IntechOpen; 2023. doi:10.5772/intechopen.106359.
21. Díez A, Carrano G, Bregón-Villahoz M, Cuétara MS, García-Ruiz JC, Fernandez-de-Larrinoa I, et al. Biomarkers for the diagnosis of invasive candidiasis in immunocompetent and immunocompromised patients. Diagn Microbiol Infect Dis 2021; 101: 115509.
22. Roberts GD, Wang HS, Hollick GE. Evaluation of the API 20 C microtube system for the identification of clinically important yeasts. J Clin Microbiol 1976; 3: 302-305.
23. Takemura H, Kaku M, Kohno S, Hirakata Y, Tanaka H, Yoshida R, et al. Evaluation of susceptibility of gram-positive and -negative bacteria to human defensins by using radial diffusion assay. Antimicrob Agents Chemother 1996; 40: 2280-2284.
24. Liang X, Yan J, Lu Y, Liu S, Chai X. The antimicrobial peptide melectin shows both antimicrobial and antitumor activity via membrane interference and DNA binding. Drug Des Devel Ther 2021; 15: 1261-1273.
25. Costa CR, Jesuíno RS, de Aquino Lemos J, de Fátima Lisboa Fernandes O, Hasimoto e Souza LK, Passos XS, et al. Effects of antifungal agents in sap activity of Candida albicans isolates. Mycopathologia 2010; 169: 91-98.
26. CLSI (2022). Performance Standards for Antifungal Susceptibility Testing of Yeasts. 3rd ed. CLSI supplement M27M44S. Replaces M60-Ed2. Clinical and Laboratory Standards Institute. https://clsi.org/media/osthhxax/m27m44sed3e_sample.pdf
27. Su L, Zhang J, Gomez H, Kellum JA, Peng Z. Mitochondria ROS and mitophagy in acute kidney injury. Autophagy 2023; 19: 401-414.
28. Grainha T, Jorge P, Alves D, Lopes SP, Pereira MO. Unraveling Pseudomonas aeruginosa and Candida albicans communication in coinfection scenarios: Insights through network analysis. Front Cell Infect Microbiol 2020; 10: 550505.
29. Hogan DA, Kolter R. Pseudomonas-Candida interactions: an ecological role for virulence factors. Science 2002; 296: 2229-2232.
30. Lindsay AK, Deveau A, Piispanen AE, Hogan DA. Farnesol and cyclic AMP signaling effects on the hypha-to-yeast transition in Candida albicans. Eukaryot Cell 2012; 11: 1219-1225.
31. Grainha T, Jorge P, Alves D, Lopes SP, Pereira MO. Unraveling Pseudomonas aeruginosa and Candida albicans Communication in coinfection scenarios: Insights through network analysis. Front Cell Infect Microbiol 2020; 10: 550505.
32. Rueda C, Puig-Asensio M, Guinea J, Almirante B, Cuenca-Estrella M, Zaragoza O, et al. Evaluation of the possible influence of trailing and paradoxical effects on the clinical outcome of patients with candidemia. Clin Microbiol Infect 2017; 23: 49.e1-49.e8.
33. Marr KA, Rustad TR, Rex JH, White TC. The trailing end point phenotype in antifungal susceptibility testing is pH dependent. Antimicrob Agents Chemother 1999; 43: 1383-1386.
34. Marcos-Zambrano LJ, Escribano P, Sánchez-Carrillo C, Bouza E, Guinea J. Scope and frequency of fluconazole trailing assessed using Eucast in invasive Candida spp. isolates. Med Mycol 2016; 54: 733-739.
35. Lee MK, Williams LE, Warnock DW, Arthington-Skaggs BA. Drug resistance genes and trailing growth in Candida albicans isolates. J Antimicrob Chemother 2004; 53: 217-224.
36. Arthington-Skaggs BA, Lee-Yang W, Ciblak MA, Frade JP, Brandt ME, Hajjeh RA, et al. Comparison of visual and spectrophotometric methods of broth microdilution MIC end point determination and evaluation of a sterol quantitation method for in vitro susceptibility testing of fluconazole and itraconazole against trailing and nontrailing Candida isolates. Antimicrob Agents Chemother 2002; 46: 2477–2481.
37. Lee MK, Kim HR, Kang JO, Kim MN, Kim EC, Kim JS, et al. Susceptibility and trailing growth of Candida albicans to fluconazole: Results of a korean multicentre study. Mycoses 2007; 50: 148-149.
38. Arthington-Skaggs BA, Warnock DW, Morrison CJ. Quantitation of Candida albicans ergosterol content improves the correlation between in vitro antifungal susceptibility test results and in vivo outcome after fluconazole treatment in a murine model of invasive candidiasis. Antimicrob Agents Chemother 2000; 44: 2081-2085.
39. Odabasi Z, Paetznick VL, Rodriguez JR, Chen E, Rex JH, Leitz GJ, et al. Lack of correlation of 24- vs. 48-h itraconazole minimum inhibitory concentrations with microbiological and survival outcomes in a Guinea pig model of disseminated candidiasis. Mycoses 2010; 53: 438-442.
40. Binder U, Aigner M, Risslegger B, Hörtnagl C, Lass-Flörl C, Lackner M. Minimal Inhibitory Concentration (MIC)-Phenomena in Candida albicans and their impact on the diagnosis of antifungal resistance. J Fungi (Basel) 2019; 5: 83.
41. Tupe SG, Kulkarni RR, Shirazi F, Sant DG, Joshi SP, Deshpande MV. Possible mechanism of antifungal phenazine-1-carboxamide from Pseudomonas sp. against dimorphic fungi Benjaminiella poitrasii and human pathogen Candida albicans. J Appl Microbiol 2015; 118: 39-48.
42. Alam F, Blackburn SA, Davis J, Massar K, Correia J, Tsai HJ, et al. Pseudomonas aeruginosa increases the susceptibility of Candida albicans to amphotericin B in dual-species biofilms. J Antimicrob Chemother 2023; 78: 2228-2241.
| Files | ||
| Issue | Vol 17 No 2 (2025) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v17i2.18395 | |
| Keywords | ||
| Antagonistic effect; Toxins of Pseudomonas aeruginosa; Candida albicans | ||
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How to Cite
1.
Hosseini MS, Navidinia M, Seyedjavadi SS, Goudarzi M, Rasouli H, Mahdavian AM, Rahimi Zamani E. Evaluation of the antagonistic effect of Pseudomonas aeruginosa toxins on azole antifungal resistance in Candida albicans species isolated from clinical samples in Iran. Iran J Microbiol. 2025;17(2):293-302.
