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Could aflatoxin B1 production by Aspergillus flavus affect the severity of keratitis: an experience in two tertiary health care centers, Egypt

  • Ghada Ibrahim Ibrahim BarakatEmail author
  • Yasmin Nabiel Kamal
  • Amira Mohammed Sultan
Original Article
  • 94 Downloads

Abstract

To investigate the expression of AFB1 gene in isolates obtained from corneal scrapping samples from keratitis patients and to correlate the quantity of AFB1 to the severity of keratitis. An observational study was undertaken in Medical Microbiology and Immunology department, Mansoura University, Egypt, over corneal scrapping samples that were cultured aiming to isolate fungal causes of infective keratitis followed by AFB1 gene detection in Aspergillus flavus isolates by nested PCR then quantitation of the toxin by TLC. Out of 843 corneal scrapping samples collected from patients with infective keratitis, positive fungal growth was identified in 277 cases (32.9%). A. flavus was the commonest fungal agent isolated in 93 cases (33.6%). The AFB1 toxin–encoding gene was detected in 63.4% of A. flavus isolates. There was a positive correlation between the quantity of produced AFB1 toxin and the degree of severity of keratitis (P value < 0.0001*). Aspergillus flavus was the most common cause of fungal keratitis, with the AFB1 toxin–encoding gene detected in more than half of the isolates. A significant correlation between the degree of severity of keratitis and the quantity of produced AFB1 toxin was detected. Therefore, exploring presence or absence of AFB1 toxin is important for the clinicians in their diagnostic assessment and selection of proper treatment choices.

Keywords

A. flavus Aflatoxin B1 Fungal keratitis Nested PCR Thin-line chromatography 

Abbreviations

A. flavus

Aspergillus flavus

AFB1

Aflatoxin B1

Nested PCR

Nested polymerase chain reaction

TLC

Thin-line chromatography

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

The study protocol acquired an approval from our institutional review board (MS.18.11.360).

Informed consent

An informative consent was received from each study participant.

References

  1. 1.
    Szczotka-Flynn LB, Pearlman E, Ghannoum M (2010) Microbial contamination of contact lenses, lens care solutions and their accessories: a literature review. Eye Contact Lens 36:116–129CrossRefGoogle Scholar
  2. 2.
    Thomas PA (2003) Fungal infections of the cornea. Eye 17:852–862CrossRefGoogle Scholar
  3. 3.
    Eaton DL, Gallagher EP (1994) Mechanisms of aflatoxin carcinogenesis. Annu Rev Pharmacol Toxicol 34:135–172CrossRefGoogle Scholar
  4. 4.
    Ehrlich KC, Yu J, Cotty PJ (2005) Aflatoxin biosynthesis gene clusters and flanking regions. J Appl Microbiol 99:518–527CrossRefGoogle Scholar
  5. 5.
    Leema G, Kaliamurthy J, Geraldine P, Thomas PA (2010) Keratitis due to Aspergillus flavus: clinical profile, molecular identification of fungal strains and detection of aflatoxin production. Mol Vis 16:843–854PubMedPubMedCentralGoogle Scholar
  6. 6.
    Chang PK (2004) Lack of interaction between AFLR and AFLJ contributes to nonaflatoxigenicity of Aspergillus sojae. J Biotechnol 107:245–253CrossRefGoogle Scholar
  7. 7.
    Hara S, Murakami H, Ohba T, Sugama S (1973) Studies on the mycological characters of aflatoxin-producing strains belonging to the A. flavus group. E. Production of moldy rice containing aflatoxin from rice and saki brewing with the moldy rice. Rep Res Inst Brew 145:13–18Google Scholar
  8. 8.
    Ahn M, Yoon K, Ryu S, Cho N, You I (2011) Clinical aspects and prognosis of mixed microbial (bacterial and fungal) keratitis. Cornea. 30:409–413CrossRefGoogle Scholar
  9. 9.
    Jones DB (1981) Decision-making in the management of microbial keratitis. Ophthalmol. 88:814–820CrossRefGoogle Scholar
  10. 10.
    Kumar M, Mishra NK, Shukla PK (2005) Sensitive and rapid polymerase chain reaction based diagnosis of mycotic keratitis through single stranded conformation polymorphism. Am J Ophthalmol 140:851–857CrossRefGoogle Scholar
  11. 11.
    Bintvihok A, Treebonmuang S, Srisakwattana K, Nuanchun W, Patthanachai K, Usawang S (2016) A rapid and sensitive detection of aflatoxin-producing fungus using an optimized polymerase chain reaction (PCR). Toxicol Res 32(1):81–87CrossRefGoogle Scholar
  12. 12.
    Przyblylski W (1975) Formation of aflatoxin derivatives on thin layer chromatographic plates. J Assoc Off Anal Chem 58:163–164PubMedGoogle Scholar
  13. 13.
    Nabney J, Nesbitt BF (1965) A spectrophotometric method for determining the aflatoxins. Analyst (Lond) 90:155–160CrossRefGoogle Scholar
  14. 14.
    Dóczi I, Gyetvai T, Kredics L, Nagy E (2004) Involvement of Fusarium spp. in fungal keratitis. Clin Microbiol Infect 10:773–776CrossRefGoogle Scholar
  15. 15.
    Zaki SM, Denning DW (2017) Serious fungal infections in Egypt. Eur J Clin Microbiol Infect Dis 36(6):971–974.  https://doi.org/10.1007/s10096-017-2929-4 CrossRefPubMedGoogle Scholar
  16. 16.
    Badawi AE, Moemen D, El-Tantawy NL (2017) Epidemiological, clinical and laboratory findings of infectious keratitis at Mansoura Ophthalmic Center. Egypt Int J Ophthalmol 10(1):61–67PubMedGoogle Scholar
  17. 17.
    Khater MM, Shehab NS, El-Badry AS (2014) Comparison of mycotic keratitis with nonmycotic keratitis: an epidemiological study. J Ophthalmol 2014:254302PubMedPubMedCentralGoogle Scholar
  18. 18.
    Bharathi MJ, Ramakrishnan R, Vasu S, Meenakshi R, Palaniappan R (2003) Epidemiological characteristics and laboratory diagnosis of fungal keratitis. A three-year study. Indian J Ophthalmol 51:315–321PubMedGoogle Scholar
  19. 19.
    Farrell S, McElnea E, Moran S, Knowles S, Murphy CC (2017) Fungal keratitis in the Republic of Ireland. Eye (Lond) 31(10):1427–1434CrossRefGoogle Scholar
  20. 20.
    Gajjar DU, Pal AK, Ghodadra BK, Vasavada AR (2013) Microscopic evaluation, molecular identification, antifungal susceptibility, and clinical outcomes in fusarium, Aspergillus and, dematiaceous keratitis. Biomed Res Int 2013:605308CrossRefGoogle Scholar
  21. 21.
    Singh G, Manikandan P, Bhaskar M, Revathi R, Narendran K, Kour A, Narendran V (2006) Multivariate analysis of childhood microbial keratitis in South India. Ann Acad Med Singap 35:185–189PubMedGoogle Scholar
  22. 22.
    Bharathi MJ, Ramakrishnan RR, Meenakshi RR, Mittal SS, Kumar CS, Srinivasan MM (2006) Microbiological diagnosis of infective keratitis: comparative evaluation of direct microscopy and culture results. Br J Ophthalmol 90:1271–1276CrossRefGoogle Scholar
  23. 23.
    Levin RE (2012) PCR detection of aflatoxin producing fungi and its limitations. Int J Food Microbiol 156:1–6CrossRefGoogle Scholar
  24. 24.
    Selvam et al. (2014) Biosci Biotech Res Asia 11(2):479–490Google Scholar
  25. 25.
    Moore GG, Singh R, Horn BW, Carbone I (2009) Recombination and lineage specific gene loss in the aflatoxin gene cluster of Aspergillus flavus. Mol Ecol 18:4870–4887CrossRefGoogle Scholar
  26. 26.
    Leema G, Chou DS, Jesudasan CA, Geraldine P, Thomas PA (2011) Expression of genes of the aflatoxin biosynthetic pathway in Aspergillus flavus isolates from keratitis. Mol Vis 17:2889–2897PubMedPubMedCentralGoogle Scholar
  27. 27.
    Flaherty JE, Payne G (1997) Overexpression of aflR leads to upregulation of pathway gene transcription and increased aflatoxin production in Aspergillus flavus. Appl Environ Microbiol 63:3995–4000PubMedPubMedCentralGoogle Scholar
  28. 28.
    Shri K, Srivastava D, Mishra RN (1984) Toxic effects of aflatoxins on eyes – an experimental clinic-histopathological evaluation. Indian J Ophthalmol 32:242–246Google Scholar
  29. 29.
    Chang P-K, Wilkinson J, Horn B, Yu J, Bhatnagar D, Cleveland T (2007) Genes differentially expressed by Aspergillus flavus strains after loss of aflatoxin production by serial transfers. Appl Microbiol Biotechnol 77:917–925CrossRefGoogle Scholar
  30. 30.
    Kim JH, Mahoney N, Chan K, Molyneux R, Campbell B (2006) Controlling food-contaminating fungi by targeting their antioxidative stress-response system with natural phenolic compounds. Appl Microbiol Biotechnol 70:735–739CrossRefGoogle Scholar
  31. 31.
    Schaafsma A, Nicol R, Savard M, Sinha R, Reid L, Rottinghaus G (1998) Analysis of Fusarium toxins in maize and wheat using thin layer chromatography. Mycopathologia 142:107–113CrossRefGoogle Scholar
  32. 32.
    Bhatnagar D, Ehrlich KC, Cleveland TE (2003) Molecular genetic analysis and regulation of aflatoxin biosynthesis. Appl Microbiol Biotechnol 61:83–93CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Faculty of MedicineMansoura UniversityMansouraEgypt

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