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Diversity, molecular phylogeny and fingerprint profiles of airborne Aspergillus species using random amplified polymorphic DNA

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Abstract

In the present study, diversity and phylogenetic relationship of Aspergillus species isolated from Tehran air was studied using random amplified polymorphic DNA (RAPD)–polymerase chain reaction (RAPD-PCR). Thirty-eight Aspergillus isolates belonging to 12 species i.e. A. niger (28.94 %, 11 isolates), A. flavus (18.42 %, 7 isolates), A. tubingensis (13.15 %, 5 isolates), A. japonicus (10.52 %, 4 isolates), A. ochraceus (10.52 %, 4 isolates), and 2.63 %, 1 isolate from each A. nidulans, A. amstelodami, A. oryzae, A. terreus, A. versicolor, A. flavipes and A. fumigatus were obtained by settle plate method which they were distributed in 18 out of 22 sampling sites examined. Fungal DNA was extracted from cultured mycelia of all Aspergillus isolates on Sabouraud Dextrose Agar and used for amplification of gene fragments in RAPD-PCR using 11 primers. RAPD-PCR data was analyzed using UPGMA software. Resulting dendrogram of combined selected primers including PM1, OPW-04, OPW-05, P160, P54, P10 and OPA14 indicated the distribution of 12 Aspergillus species in 8 major clusters. The similarity coefficient of all 38 Aspergillus isolates ranged from 0.02 to 0.40 indicating a wide degree of similarities and differences within and between species. Taken together, our results showed that various Aspergillus species including some important human pathogenic ones exist in the outdoor air of Tehran by different extents in distribution and diversity and suggested inter- and intra-species genetic diversity among Aspergillus species by RAPD-PCR as a rapid, sensitive and reproducible method.

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References

  • Abed KF (2008) Differentiation of Aspergillus niger by random amplification of polymorphic DNA. J Ind Microbiol Biotechnol 35:1027–1032

    Article  CAS  Google Scholar 

  • Abu Seadah AA, EL Shikh ME (2008) RAPD typing of Aspergillus chevalieri, Aspergillus nidulans, Aspergillus tetrazonus (quadrilineatus) and their teleomorphs using 5′-d[AACGCGCAAC]-3′ and 5′-d[CCCGTCAGCA]-3′ primers. Mol Biol Rep 35:89–95

    Article  CAS  Google Scholar 

  • Aiat N (2006) Genetic variability among three species of Aspergillus 2. Random amplified polymorphic DNA (RAPD) markers for genetic analysis. J Appl Sci Res 2:709–713

    Google Scholar 

  • Andon BM (2006) Active air vs. passive air (settle plate) monitoring in routine environmental monitoring programs. J Pharm Sci Technol 60:350–355

    Google Scholar 

  • Araujo R, Cabral JP (2010) Fungal air quality in medical protected environments. In: Kumar A (ed) Air quality., 17InTech Open Access Publisher, Croatia, pp 357–382

    Google Scholar 

  • Bakri Y, Arabi MI, Jawhar M (2007) RAPD technique is a useful tool to distinguish Penicillium species. Pol J Microbiol 56:273–276

    CAS  Google Scholar 

  • Batista PP, Santos JF, Liveira NT, Pires APD, Motta CMS, Luna-Alves Lima EA (2008) Genetic characterization of Brazilian strains of Aspergillus flavus using DNA markers. Genet Mol Res 7:706–717

    Article  CAS  Google Scholar 

  • Baysal O, Siragusa M, Gumrukcu E, Zengin S, Carimi F, Sajeva M, Teixeira da Silva JA (2010) Molecular characterization of Fusarium oxysporum f. melongenae by ISSR and RAPD markers on egg planet. Biochem Genet 48:524–537

    Article  CAS  Google Scholar 

  • Bhattacharya E, Ranade SA (2001) Molecular distinction amongst varieties of Mulberry using RAPD and DAMD profiles. BMC Planet Biol 1:3

    Article  CAS  Google Scholar 

  • Calvo A, Guarro J, Suarez G, Ramirez C (1980) Air-borne fungi in the air of Barcelona (Spain). III: the genus aspergillus link. Mycopathologia 71:41–43

    Article  CAS  Google Scholar 

  • de Valk HA, Meis JF, Curfs IM, Muehlethaler K, Mouton JW, Klaassen CH (2005) Use of a novel panel of nine short tandem repeats for exact and high-resolution fingerprinting of Aspergillus fumigatus isolates. J Clin Microbiol 43:4112–4120

    Article  Google Scholar 

  • Denning DW (1998) Invasive aspergillosis. Clin Infect Dis 26:781–805

    Article  CAS  Google Scholar 

  • Diaz-Guerra TM, Mellado E, Cuenca-Estrella M, Gaztelurrutia L, Navarro JIV, Tudela JLR (2000) Genetic similarity among one Aspergillus flavus strain isolated from a patient who underwent heart surgery and two environmental strains obtained from the operating room. J Clin Microbiol 38:2419–2422

    CAS  Google Scholar 

  • Diba K, Makhdoomi K, Mirhendi H (2014) Molecular characterization of Aspergillus infections in an Iranian educational hospital using RAPD-PCR method. Iran J Basic Med Sci 17:646

    CAS  Google Scholar 

  • Fungaro MHP, Magnani M, Vilas-Boas LA, Vissotto PC, Furlaneeto MC, Vieira MLC, Taniwaki MH (2004) Genetic relationships among Brazilian strains of Aspergillus ochraceus based on RAPD and ITS sequences. Can J Microbiol 50:985–988

    Article  CAS  Google Scholar 

  • Gehlot P, Purohit DK, Singh SK (2011) Molecular diagnostics of human pathogenic Aspergillus species. Indian J Biotechnol 10:207–211

    CAS  Google Scholar 

  • Guinea J, Peláez T, Alcalá L, Bouza E (2005) Evaluation of Czapek agar and Sabouraud dextrose agar for the culture of airborne Aspergillus conidia. Diagn Microbiol Infect Dis 53:333–334

    Article  CAS  Google Scholar 

  • Hedayati MT, Mayahi S, Aghili R, Goharimoghadam K (2005) Airborne fungi in indoor and outdoor of asthmatic patients’ home, living in the city of Sari. Iran J Allerg Asthma Immunol 4:189–191

    Google Scholar 

  • Hoekstra ES, Samson RA, Summerbell R. (2000) Introduction to food– and airborne fungi, Samson RA (ed), Baarn: Centraalbureau voor Schimmelcultures; The Netherlands

  • Irshad S, Nawab R (2012) Molecular characterization of seven different species of Aspergillus through random amplified polymorphic DNA (RAPD) and enzyme analysis. J Microbiol Res 2:47–50

    Article  Google Scholar 

  • Jamali M, Ebrahimi MA, Karimipour M, Shams-Ghahfarokhi M, Dinparast-Djadid M, Kalanetari S, Pilehvar-Soltanahmadi Y, Amani A, Razzaghi-Abyaneh M (2012) An insight into the distribution, genetic diversity, and mycotoxin production of Aspergillus section Flavi in soils of pistachio orchards. Folia Microbiol 57:27–36

    Article  CAS  Google Scholar 

  • Khan ZK, Chowdhary L, Gyanchandani A, Goswami M, Farooqui N, Ranade SA (1998) Aspergillus fumigatus strains recovered from immunocompromised patients (ICP): subtyping of strains by RAPD analysis. Biochem Mol Biol Int 46:537–548

    CAS  Google Scholar 

  • Kim KY, Kim YS, Kim D (2010) Distribution characteristics of airborne bacteria and fungi in the general hospitals of Korea. Ind Health 48:236–243

    Article  Google Scholar 

  • Loo VG, Bertrand C, Dixon C (1996) Control of construction-associated nosocomial Aspergillosis in an antiquated hematology unit. Infect Control Hosp Epidemiol 17:360–364

    Article  CAS  Google Scholar 

  • Loudon KW, Burine J, Coke AP, Matthews RC (1993) Application of polymerase chain reaction to fingerprinting by random amplification of polymorphic DNA. J Clin Microbiol 31:1117–1121

    CAS  Google Scholar 

  • Lourenço A, Durigon EL, Zanotto P, Maderia JEGC, Almeida APD, Correa B (2007) Genetic diversity of environmental Aspergillus flavus strains in the state of São Paulo, Brazil by random amplified polymorphic DNA. Mem Inst Oswaldo Cruz 102:687–692

    Article  Google Scholar 

  • Mallea M, Murray IG, Segretain G, Philpot CM, Charpin H, Gueho E, Charpin J (1972) Census of Aspergillus colonies in the air comparison between London, Paris, Lyon, Marseilles. Acta Allergol 127:273–278

    Article  Google Scholar 

  • Maslow JN, Mulligan ME, Arbeit RD (1993) Molecular epidemiology: application of contemporary techniques to the typing of microorganisms. Clin Infect 17:153–164

    Article  CAS  Google Scholar 

  • Melchers WJ, Verweij PE, Van Den Hurk P, Van Belkum A, De Pauw BE, Hoogkamp-Korstanje JA, Meis JF (1994) General primer-mediated PCR for detection of Aspergillus species. J Clin Microbiol 32:1710–1717

    CAS  Google Scholar 

  • Menotti J, Waller J, Meunier O (2005) Epidemiological study of invasive pulmonary aspergillosis in a heamatology unit by molecular typing of environmental and patient isolates of Aspergillus fumigatus. J Hosp Infect 60:61–68

    Article  CAS  Google Scholar 

  • Midorikawa GEO, Pinheiro MRR, Vidigal BS, Arruda MC, Costa FF, Pappas GJ Jr, Miller RNG (2008) Characterization of Aspergillus flavus strains from Brazilian nuts and cashew by RAPD and ribosomal DNA analysis. Lett Appl Microbiol 47:12–18

    Article  CAS  Google Scholar 

  • Narasimhan B, Asokan M (2010) Genetic variability of Aspergillus terreus from dried grapes using RAPD-PCR. Adv Biosci Biotechnol 1:345–353

    Article  Google Scholar 

  • NDSC. National guidelines for the prevention of nosocomial invasive Aspergillosis during construction/renovation activities. Developed by a sub-committee of the scientific advisory committee of the national disease surveillance centre 2002, ISBN 0-9540177-3-0

  • Opal SM, Asp AA, Cannady PB Jr, Morse PL, Burton LJ, Hammer PG II (1986) Efficacy of infection control measures during a nosocomial outbreak of disseminated aspergillosis associated with hospital construction. J Infect Dis 153:634–637

    Article  CAS  Google Scholar 

  • Pakshir K, Shekarkhar G, Mostagnie S, Sabayan B, Vaghefikia A (2007) Monitoring of airborne fungi in two general hospitals in Shiraz, Southern Iran. Iran J Med Sci 32:240–244

    Google Scholar 

  • Panagopoulou P, Filioti J, Petrikkos G, Giakouppi P, Anatoliotaki M, Farmaki E, Kanta A, Apostolakou H, Avlami A, Samonis G, Roilides E (2002) Environmental surveillance of filamentous fungi in three tertiary care hospitals in Greece. J Hosp Infect 52:185–191

    Article  CAS  Google Scholar 

  • Pavlícek A, Hrdá S, Flegr J (1999) Free-Tree-freeware program for construction of phylogenetic trees on the basis of distance data and bootstrap/jackknife analysis of the tree robustness. Application in the RAPD analysis of genus Frenkelia. Folia Biol 45:97–99

    Google Scholar 

  • Raclaský V, Trtková J, Rusková L, Buchta V, Bolehovská R, Vacková M, Hamal P (2006) Primer R108 performs best in the RAPD strain typing of three Aspergillus species frequently isolated from patients. Folia Microbiol 51:136–140

    Article  Google Scholar 

  • Raclavský V, Trtkova J, Ruskova L, Buchta V, Bolehovska R, Vackova M, Hamal P (2006) Primer R108 performs best in the RAPD strain typing of three Aspergillus species frequently isolated from patients. Folia Microbiol 51:136–140

    Article  Google Scholar 

  • Raper KB, Fennel DI (1965) The Genus Aspergillus. Williams & Wilkins, Baltimore

    Google Scholar 

  • Rath PM, Ansorg R (1997) Value of environmental sampling and molecular typing of Aspergilli to assess nosocomial sources of aspergillosis. J Hosp Infect 37:47–53

    Article  CAS  Google Scholar 

  • Rath PM, Kamphoff S, Ansorg R (1999) Value of different methods for the characterization of Aspergillus terreus strains. J Med Microbiol 48:161–166

    Article  CAS  Google Scholar 

  • Rath PM, Petermeier K, Verweij PE, Ansorg R (2002) Differentiation of Aspergillus ustus Strains by Random Amplification of Polymorphic DNA. J Clin Microbiol 40:2231–2233

    Article  CAS  Google Scholar 

  • Razzaghi-Abyaneh M, Shams-Ghahfarokhi M, Chang P-K (2011) Aflatoxins: mechanisms of inhibition by antagonistic planets and microorganisms. In: Ramon G. Guevara-Gonzalez (ed) Aflatoxins: Biochemistry and Molecular Biology, INTECH Open Access Publisher, pp 285–304

  • Sarubbi FA Jr, Kopf HB, Wilson MB, McGinnis MR, Rutala WA (1982) Increased recovery of Aspergillus flavus from respiratory specimens during hospital construction. Am Rev Respir Dis 125:33–38

    Google Scholar 

  • Sautour M, Sixt N, Dalle F, L’Ollivier C, Fourquenet V, Calinon C, Paul K, Valvin S, Maurel A, Aho S, Couillault G, Cachia C, Vagner O, Cuisenier B, Caillot D, Bonnin A (2009) Profiles and seasonal distribution of airborne fungi in indoor and outdoor environments at a French hospital. Sci Total Environ 407:37666–37671

    Article  Google Scholar 

  • Schmidt H, Taniwaki MH, Vogel RF, Iessen L (2004) Utilization of AFLP markers for PCR-based identification of Aspergillus carbonarius and indication of its presence in green coffee samples. J App Microbiol 97:899–909

    Article  CAS  Google Scholar 

  • Sepahvand A, Shams-Ghahfarokhi M, Allameh A, Jahanshiri Z, Jamali M, Razzaghi-Abyaneh M (2011) A survey on distribution and toxigenicity of Aspergillus flavus from indoor and outdoor hospital environments. Folia Microbiol 56:527–534

    Article  CAS  Google Scholar 

  • Sepahvand A, Shams-Ghahfarokhi M, Allameh A, Razzaghi-Abyaneh M (2013) Diversity and distribution patterns of airborne microfungi in indoor and outdoor hospital environments in Khorramabad, Southwest Iran. Jundishapur J Microbiol 6:186–192

    Article  Google Scholar 

  • Shams-Ghahfarokhi M, Aghaei-Gharehbolagh S, Aslani N, Razzaghi-Abyaneh M (2014) Investigation on distribution of airborne fungi in outdoor environment in Tehran, Iran. J Environ Health Sci Engin 12:54

    Article  Google Scholar 

  • Symoens F, Bouchara JP, Heinemann S, Nolard N (2000) Molecular typing of Aspergillus terreus isolates by random amplification of polymorphic DNA. J Hosp Infect 44:273–280

    Article  CAS  Google Scholar 

  • Turenne CY, Sanche SE, Hoban DJ, Karlowsky JA, Kabani AM (1999) Rapid identification of fungi by using the ITS2 genetic region and an automated fluorescent capillary electrophoresis system. J Clin Microbiol 37:1846–1851

    CAS  Google Scholar 

  • Vonberg RP, Gastmeier P (2006) Nosocomial aspergillosis in outbreak settings. J Hosp Infect 63:246–254

    Article  Google Scholar 

  • Wiederhold NP, Lewis RE, Kontoyiannis DP (2003) Invasive aspergillosis in patients with hematologic malignancies. Pharmacogn Ther 23:1592–1610

    Google Scholar 

  • Zaini F, Hedayati MT (1995) Study of airborne fungi in the wards of 3 Tehran hospitals. J Med Council Islam Repub Iran 13:17–20

    Google Scholar 

Download references

Acknowledgments

This work was financially supported by Tarbiat Modares University. The authors wish to thank Mrs. Razeghi for helpful technical assistance.

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Authors and Affiliations

  1. Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, 14115-331, Iran

    Firoozeh Kermani & Masoomeh Shams-Ghahfarokhi

  2. Department of Mycology, Pasteur Institute of Iran, Tehran, 13164, Iran

    Mohammadhassan Gholami-Shabani & Mehdi Razzaghi-Abyaneh

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  1. Firoozeh Kermani
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  2. Masoomeh Shams-Ghahfarokhi
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Correspondence to Masoomeh Shams-Ghahfarokhi.

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Kermani, F., Shams-Ghahfarokhi, M., Gholami-Shabani, M. et al. Diversity, molecular phylogeny and fingerprint profiles of airborne Aspergillus species using random amplified polymorphic DNA. World J Microbiol Biotechnol 32, 96 (2016). https://doi.org/10.1007/s11274-016-2052-1

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