Molecular characterization and strain typing of fungal contaminants of Processed Manihot esculenta Crantz (“Garri”) in Ogun State, Nigeria

Authors

  • B T Thomas Olabisi Onabanjo University
  • L A Ogunkanmi
  • B A Iwalokun
  • G C Agu

Keywords:

Aspergillus species, Electrophoresis, Fungi, Genetic diversity, Processed Manihot esculenta (Garri), Random Amplified Polymorphic DNA (RAPD)

Abstract

Background: The lack of standard biochemical tests, coupled with the limitation of the number of micro and macro morphology that can be scored for fungal diagnosis, makes the identification of fungal organisms using microbiological methods less reliable.

Objective: To determine the fungal contaminants of processed Manihot esculenta Crantz (Garri) and provide information on their diversity.

Methods: In this study, fungal contaminants of processed Manihot esculenta Crantz (Garri) were characterized by sequencing the hyper-variable 18S ribosomal RNA as well as type the isolated organisms using Random Amplified Polymorphic DNA markers.

Results: The  PCR amplification of the 18S ribosomal RNA gene of the  isolated fungi from  processed Manihot esculenta Crantz yielded a single fragment of an approximately between 600 and 700 bp. BLAST search using Genbank database showed that the isolate percentage similarity with GenBank accessions ranged between 98% and 100%. The molecular technique successfully identified all the isolated organisms resulting in 100% accurate diagnosis as against 79.3%accurate diagnosis made with the microbiological method. The forty isolated Aspergillus species were further resolved into 27 RAPD haplotypes with Simpson Index of Genetic Diversity approaching one for all the isolates. The mean genetic diversity within (GL) and among (GS) the isolated Aspergillus species were found to be 89% and 11% respectively. The total genetic diversity (HT) for these organisms was approximately 48%.  These results connoted significant strain diversity in the sampled specimens as shown by differences in their electrophoretic patterns.

Conclusion: The study revealed significant strain diversity in the sampled specimens as shown by differences in their electrophoretic patterns.

Author Biography

B T Thomas, Olabisi Onabanjo University

Microbiology/Lecturer II

References

Ikediobi CO, Onfia GOC, Eluwah CE. A rapid and inexpensive enzymatic assay for total cyanide in cassava (Manihot esculenta crantz) cassava products. Agri Bio Chem 1980; 44: 2803-09.

Oluwole OB, Olatunji OO, Odunfa SA. A process technology for conservation of dried cassava chips with garri. Niger Food J 2004; 22: 65-73.

Jekayinfa SO, Olajide JO. Analysis of energy usage in the production of three selected cassava based foods in Nigeria. J Food Eng 2007; 82: 217-26.

Ogiehor IS, Ikenebomeh MJ, Ekundayo AO. The bioload and aflatoxin content of market garri from some selected states in southern Nigeria: public health significance. Afr Health Sci 2007; 7(4): 223-27.

Arisa NU, Omosaiye OB, Adelekan AO, Alabi-Mac FA. Chemical and sensory qualities of gari fortified with groundnut flour. Afr J Food SciTechnol 2011; 2(5): 116-19.

Osho SM. The processing and acceptability of a fortified cassava-based product (gari) with soybean. Nutri Food Sci 2003; 33(6): 278-83.

Thomas BT, Effedua HI, Agu GC, Musa OS, Adeyemi MT, Odunsi OD, et al. Fungi associated with the deterioration of garri in Ogun State, Nigeria. Reseacher 2012; 4(2): 8-12.

International Commission of Microbiological Specification for Food (ICMSF). Microorganisms in foods. Kluwer academic/plenum. 2002Vol 7. USA.

Larone DH. Medically important fungi: a guide to identification. 4th Edition. Washington D.C, ASM Press. 2002: pp. 150 – 310.

Fredricks D, Smith C, Meier A. Comparison of six DNA extraction methods for recovery of fungal DNA as assessed by quantitative PCR. J Clin Microbiol 2005; 43(10): 5122-28.

Pryce TM, Palladino S, Kay ID, Coombs GW. Rapid identification of fungi by sequencing the ITS1 and ITS2 regions using an automated capillary electrophoresis system. Med Mycol 2003; 41: 369-81.

Roodt R, Spies JJ Burger TH. Preliminary DNA fingerprinting of the turfgrass Cynodon dactylon (Poaceae: Chloridoideae). Bothalia 2002; 32: 117-22.

Tamura K, Dudley J, Nei M, Kumar S. MEGA 4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Molecular Biol Evolution 2007; 24: 1596-99. Available at http://www.kumarlab.net/publications)

Hammer O, Harper DAT, Ryan PD. PAST: Paleontological statistic software package for education and data analysis. Paleontologia Eletronica 2001; 4(1): 1-9. Available at: http://palaeoelectronica.org/2001 _1/past/issue1_01.htm [Accessed: 04/06/ 2015]

Bhattacharya S, Mondal AS. Clinical microbiology in the intensive care unit: strategic and operational characteristics. Ind J Med Microbiol 2010; 28(1): 5-10.

Schwan RF, Wheals AE. The microbiology of cocoa fermentation and its role in chocolate quality. Crit Rev Food Sci Nutri 2004; 44: 205–21.

Ogiehor IS, Ikenebomeh MJ. The effect of different packaging materials on the shelf stability of garri. Afr J Biotechnol 2005; 523: 2412-16.

Magnolia C, Hallak C, Astoreeca A, Ponsone L, Chiazchiera S, Dalcero AM. Occurrence of ochratoxin A producing fungi in commercial kernels in Argentina. Mycopathologia 2006; 161: 53-8.

Zimmerli B, Dick R. Ochratoxin A in table wine and grape-juice: occurrence and risk wine and its geographical origin. Food Additives and Contaminations 1996; 17: 793–98.

Otteneder H, Majerus P. Occurrence of ochratoxin A (OTA) in wines: influence of the type of wine and its geographical origin. Food Additives and Contaminations 2000; 17: 793–98.

Samson RA. Current fungal taxonomy and mycotoxins. Mycotoxins and phycotoxins in perspective at the turn of the millennium. In: Proceedings of the Xth International IUPAC Symposium on Mycotoxins and Phycotoxins, 21-25 May, 2000. Guarujá (Brazil). Ponsen & Looyen, Wageningen, The Netherlands.

Bennett JW, Klich J. Mycotoxins. Clin Microbiol Rev 2003; 16(3): 497.

Cheesebrough LM. Medical Laboratory Manual for Tropical Countries (Volume II). London, UK. Butterworth & Co. Publishers. 2005.

Pekarek E, Jacobson K,. Donovan A. High levels of genetic variation exist in Aspergillus niger populations infecting Welwitschia mirabilis hook. J Hered 2006; 97(3): 270-78.

Grypta H, Debaud JC, Marmeise R. Population dynamics of the symbiotic mushroom Hebeloma cylindrosporum: mycelial persistence and inbreeding. Heredity 2000; 84: 294–302.

Downloads

Additional Files

Published

2017-12-01

Issue

Section

Articles