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Abstract

Purpose. Non-albicans Candida species have emerged as fungal pathogens that cause invasive infections, with many of these species displaying resistance to commonly used antifungal agents. This study was confined to studying the characteristics of clinical isolates of the C. rugosa complex and C. pararugosa species.

Methodology. Seven isolates of the C. rugosa complex and one isolate of C. pararugosa were obtained from two tertiary referral hospitals in Malaysia. Their antifungal susceptibilities, biofilm, proteinase, phospholipase, esterase and haemolysin activities were characterized. Biofilms were quantified using crystal violet (CV) and tetrazolium (XTT) reduction assays at 1.5, 6, 18, 24, 48 and 72 h.

Results/Key findings. The E-test antifungal tests showed that both species have elevated MICs compared to C. albicans and C. tropicalis. The highest biomass was observed in one of the C. rugosa isolates (0.237), followed by C. pararugosa (0.206) at 18 h of incubation. However, the highest bioactivity was observed in the C. rugosa ATCC 10571 strain at 24 h (0.075), followed by C. pararugosa at 48 h (0.048) and the same C. rugosa strain at 24 h (0.046), with P<0.05. All isolates exhibited high proteinase activity (+++) whereas six isolates showed very strong esterase activity (++++). All the isolates were alpha haemolytic producers. None of the isolates exhibited phospholipase activity.

Conclusion. Elevated MICs were shown for the C. rugosa complex and C. pararugosa for commonly used antifungal drugs. Further studies to identify virulence genes involved in the pathogenesis and genes that confer reduced drug susceptibility in these species are proposed.

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2019-02-06
2019-10-18
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References

  1. Arzmi MH, Alshwaimi E, Harun WHAW, Razak FA, Farina F et al. Gaining more insight into the determinants of Candida species pathogenicity in the oral cavity. Eur J Inflamm 2014;12:227–235 [CrossRef]
    [Google Scholar]
  2. Singh S, Nawange SR, Warthe N. In-vitro antifungal susceptibility reveals occurrence of Azole and Allylamine resistance among clinical isolates of Candida albicans and Candida non albicans from central India. Int J Pharm Sci Res 2014;5:5267–5275
    [Google Scholar]
  3. Jiang C, Dong D, Yu B, Cai G, Wang X et al. Mechanisms of azole resistance in 52 clinical isolates of Candida tropicalis in China. J Antimicrob Chemother 2013;68:778–785 [CrossRef]
    [Google Scholar]
  4. Singh RI, Xess I, Mathur P, Behera B, Gupta B et al. Epidemiology of candidaemia in critically ill trauma patients: experiences of a level I trauma centre in North India. J Med Microbiol 2011;60:342–348 [CrossRef]
    [Google Scholar]
  5. Oberoi JK, Wattal C, Goel N, Raveendran R, Datta S et al. Non-albicans Candida species in blood stream infections in a tertiary care hospital at New Delhi, India. Indian J Med Res 2012;136:997–1003 PMCID: PMC3612330
    [Google Scholar]
  6. Yang YL, Lin CC, Chang TP, Lauderdale TL, Chen HT et al. Comparison of human and soil Candida tropicalis isolates with reduced susceptibility to fluconazole. PLoS One 2012;7:e34609 [CrossRef]
    [Google Scholar]
  7. Nalina K. Candida infections: novel virulence factors and mechanisms of azole resistance. Malays J Microbiol 2014;10:294–303
    [Google Scholar]
  8. Amran F, Aziz MN, Ibrahim HM, Atiqah NH, Parameswari S et al. In vitro antifungal susceptibilities of Candida isolates from patients with invasive candidiasis in Kuala Lumpur Hospital, Malaysia. J Med Microbiol 2011;60:1312–1316 [CrossRef]
    [Google Scholar]
  9. Ding CH, Wahab AA, Muttaqillah NA, Tzar MN. Prevalence of albicans and non-albicans candiduria in a Malaysian medical centre. J Pak Med Assoc 2014;64:1375–1379 PMID: 25842581
    [Google Scholar]
  10. Chakravarthi S, Haleagrahara N. A comprehensive review of the occurrence and management of systemic candidiasis as an opportunistic infection. Microbiol J 2011;1:1–7 [CrossRef]
    [Google Scholar]
  11. Paredes K, Sutton DA, Cano J, Fothergill AW, Lawhon SD et al. Molecular identification and antifungal susceptibility testing of clinical isolates of the Candida rugosa species complex and proposal of the new species Candida neorugosa. J Clin Microbiol 2012;50:2397–2403 [CrossRef]
    [Google Scholar]
  12. Chaves GM, Terçarioli GR, Padovan ACB, Rosas RC, Ferreira RC et al. Candida mesorugosa sp. nov., a novel yeast species similar to Candida rugosa, isolated from a tertiary hospital in Brazil. Med Mycol 2013;51:231–242 [CrossRef]
    [Google Scholar]
  13. Padovan AC, Melo AS, Colombo AL. Systematic review and new insights into the molecular characterization of the Candida rugosa species complex. Fungal Genet Biol 2013;61:33–41 [CrossRef]
    [Google Scholar]
  14. Khunnamwong P, Lertwattanasakul N, Jindamorakot S, Limtong S, Lachance MA. Description of Diutina gen. nov., Diutina siamensis, f.a. sp. nov., and reassignment of Candida catenulata, Candida mesorugosa, Candida neorugosa, Candida pseudorugosa, Candida ranongensis, Candida rugosa and Candida scorzettiae to the genus Diutina. Int J Syst Evol Microbiol 2015;65:4701–4709 [CrossRef]
    [Google Scholar]
  15. Papon N, Courdavault V, Clastre M, Bennett RJ. Emerging and emerged pathogenic Candida species: beyond the Candida albicans paradigm. PLoS Pathog 2013;9:e1003550 [CrossRef]
    [Google Scholar]
  16. Adjapong G, Bartlett M, Hale M, Garrill A. The isolation of Candida rugosa and Candida mesorugosa from clinical samples in Ghana. Med Myco 2016;54:322–326 [CrossRef]
    [Google Scholar]
  17. Tay ST, Tan HW, Na SL, Lim SL. Phenotypic and genotypic characterization of two closely related subgroups of Candida rugosa in clinical specimens. J Med Microbiol 2011;60:1591–1597 [CrossRef]
    [Google Scholar]
  18. Ng KP, Kuan CS, Kaur H, Na SL, Atiya N et al. Candida species epidemiology 2000-2013: a laboratory-based report. Trop Med Int Health 2015;20:1447–1453 [CrossRef]
    [Google Scholar]
  19. Taj-Aldeen SJ, AbdulWahab A, Kolecka A, Deshmukh A, Meis JF et al. Uncommon opportunistic yeast bloodstream infections from Qatar. Med Mycol 2014;52:552–556 [CrossRef]
    [Google Scholar]
  20. Weerasekera MM, Sissons CH, Wong L, Anderson S, Holmes AR et al. Use of denaturing gradient gel electrophoresis for the identification of mixed oral yeasts in human saliva. J Med Microbiol 2013;62:319–330 [CrossRef]
    [Google Scholar]
  21. Seifi Z, Zarei Mahmoudabadi A, Zarrin M. Extracellular enzymes and susceptibility to fluconazole in Candida strains isolated from patients with vaginitis and healthy individuals. Jundishapur J Microbiol 2015;8:e20162 [CrossRef]
    [Google Scholar]
  22. Fatahinia M, Halvaeezadeh M, Rezaei-Matehkolaei A. Comparison of enzymatic activities in different Candida species isolated from women with vulvovaginitis. J Mycol Med 2017;27:188–194 [CrossRef]
    [Google Scholar]
  23. Kaur R, Goyal R, Dhakad MS, Bhalla P, Diwan R. A study of virulence factors: proteinase, phospholipase, and biofilm in HIV/ AIDS patients. J HIV AIDS 2015;1:1–7
    [Google Scholar]
  24. Bassyouni RH, Wegdan AA, Abdelmoneim A, Said W, AboElnaga F. Phospholipase and aspartyl proteinase activities of Candida species causing vulvovaginal candidiasis in patients with Type 2 diabetes mellitus. J Microbiol Biotechnol 2015;25:1734–1741 [CrossRef]
    [Google Scholar]
  25. White TJ, Bruns T, Lee S, Taylor J.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics In MA Innis, DH Gelfand, JJ Sninsky, TJ White. (editors) PCR Protocols: A Guide to Methods and Applications New York, USA: Academic Press; 1990;315–322
    [Google Scholar]
  26. Pierce CG, Uppuluri P, Tristan AR, Wormley FL, Mowat E et al. A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing. Nat Protoc 2008;3:1494–1500 [CrossRef]
    [Google Scholar]
  27. Pierce CG, Uppuluri P, Tummala S, Lopez-Ribot JL. A 96 well microtiter plate-based method for monitoring formation and antifungal susceptibility testing of Candida albicans biofilms. J Vis Exp 2010;2010:e2287 [CrossRef]
    [Google Scholar]
  28. O’Toole GA. Microtiter dish biofilm formation assay. J Vis Exp 2011;2437 [CrossRef]
    [Google Scholar]
  29. Kumar DA, Muralidhar S, Biswas K, Banerjee U, Basir SF et al. Species diversity, antifungal susceptibility, and virulence attributes of Candida colonizing the oral cavities of adult diabetic patients. J Mycol 2014;9:395041
    [Google Scholar]
  30. Vinodhini R, Moorthy K, Suresh M. Incidence and virulence traits of candida dubliniensis isolated from clinically suspected patients. Asian J Pharm Clin Res 2016;9:77–81 [CrossRef]
    [Google Scholar]
  31. Goyal RK, Sami H, Mishra V, Bareja R, Behara RN. Non-albicans Candiduria: an emerging threat. J App Pharm Sci 2016;6:048–050 [CrossRef]
    [Google Scholar]
  32. Sheikh N, Jahagirdar V, Kothadia S, Nagoba B. Antifungal drug resistance in Candida species. Eur J Gen Med 2013;10:254–258 [CrossRef]
    [Google Scholar]
  33. Chaurasia D, Shrivastava RK, Dubey D. Characterization and antifungal susceptibility pattern of Candida isolated from blood stream infections among patients admitted in NICU and PICU of a tertiary care centre. Int J Sci Technol 2015;4:174–177
    [Google Scholar]
  34. Madhavan P, Jamal F, Chong PP, Ng KP. In vitro activity of fluconazole and voriconazole against clinical isolates of Candida spp. by E-test method. Trop Biomed 2010;27:200–207 PMID: 20962716
    [Google Scholar]
  35. Minces LR, Ho KS, Veldkamp PJ, Clancy CJ. Candida rugosa: a distinctive emerging cause of candidaemia. A case report and review of the literature. Scand J Infect Dis 2009;41:892–897 [CrossRef]
    [Google Scholar]
  36. Bitar I, Khalaf RA, Harastani H, Tokajian S. Identification, typing, antifungal resistance profile, and biofilm formation of Candida albicans isolates from Lebanese hospital patients. Biomed Res Int 2014;2014:1–10 [CrossRef]
    [Google Scholar]
  37. Karimi L, Mirhendi H, Khodadadi H, Mohammadi R. Molecular identification of uncommon clinical yeast species in Iran. Curr Med Mycol 2015;1:1–6 [CrossRef]
    [Google Scholar]
  38. Capoor MR, Gupta DK, Verma PK, Sachdeva HC. Rare yeasts causing fungemia in immunocompromised and haematology patients: case series from Delhi. Indian J Med Microbiol 2015;33:576–579 [CrossRef]
    [Google Scholar]
  39. Pfaller MA, Diekema DJ, Gibbs DL, Newell VA, Ellis D et al. Results from the ARTEMIS DISK Global antifungal Surveillance study, 1997 to 2007: a 10.5-year analysis of susceptibilities of Candida species to fluconazole and voriconazole as determined by CLSI standardized disk diffusion. J Clin Microbiol 2010;48:1366–1377 [CrossRef]
    [Google Scholar]
  40. Pfaller MA, Diekema DJ, Messer SA, Boyken L, Hollis RJ et al. In vitro susceptibilities of rare Candida bloodstream isolates to ravuconazole and three comparative antifungal agents. Diagn Microbiol Infect Dis 2004;48:101–105 [CrossRef]
    [Google Scholar]
  41. Aslam A, Akhtar N, Hasan F, Shah AA. Prevalence and in vitro antifungal susceptibility pattern of Candida species in a tertiary care hospital, Rawalpindi, Pakistan. Pak J Zool 2015;47:335–342
    [Google Scholar]
  42. Sugar AM, Stevens DA. Candida rugosa in immunocompromised infection case reports, drug susceptibility, and review of the literature. Cancer 1985;56:318–320 [CrossRef]
    [Google Scholar]
  43. Kocyigit I, Unal A, Sipahioglu MH, Tokgoz B, Oymak O et al. Peritonitis due to Candida rugosa: the first case report. Perit Dial Int 2010;30:576–577 [CrossRef]
    [Google Scholar]
  44. Hamid SU, Tan S, Ridzuan SNA, Seman MSC, Ramli R et al. Antifungal susceptibility patterns among candida species isolated from blood at Universiti Kebangsaan Malaysia Medical Centre. Sains Malays 2012;41:961–967
    [Google Scholar]
  45. Verma S, Angrup A, Sharma V, Shivaprakash MR. Candida rugosa candidaemia in a critically III trauma patient successfully treated with amphotericin B. J med sci clin res 2017;5:28930–28933
    [Google Scholar]
  46. Shirkhani S, Sepahvand A, Mirzaee M, Anbari K. Phospholipase and proteinase activities of Candida spp. isolates from vulvovaginitis in Iran. J Mycol Med 2016;26:255–260 [CrossRef]
    [Google Scholar]
  47. Luo G, Samaranayake LP, Yau JYY. Candida species exhibit differential in vitro hemolytic activities. J Clin Microbiol 2001;39:2971–2974 [CrossRef]
    [Google Scholar]
  48. Butola R, Agwan V, Thakuria B, Madan M. A comparative study of virulence factors in clinical isolates of Candida species. Int J Curr Microbiol Appl Sci 2015;4:716–722
    [Google Scholar]
  49. Turan H, Demirbilek M. Biofilm-forming capacity of blood–borne Candida albicans strains and effects of antifungal agents. Rev Argent Microbiol 2018;50:62–69 [CrossRef]
    [Google Scholar]
  50. Majumdar T, Mullick JB, Bir R, Roy J, Sil SK. Determination of virulence factors and biofilm formation among isolates of vulvovaginal candidiasis. J Med Sci 2016;36:53–58 [CrossRef]
    [Google Scholar]
  51. Martins CHG, Pires RH, Cunha AO, Pereira CAM, Singulani JL et al. Candida/Candida biofilms. first description of dual-species Candida albicans/C. rugosa biofilm. Fungal Biol 2016;120:530–537 [CrossRef]
    [Google Scholar]
  52. Mendes A, Mores AU, Carvalho AP, Rosa RT, Samaranayake LP et al. Candida albicans biofilms produce more secreted aspartyl protease than the planktonic cells. Biol Pharm Bull 2007;30:1813–1815 [CrossRef]
    [Google Scholar]
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