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Molecular Identification of Black Aspergilli Isolated from Poultry Feeds by Sequencing their ITS-Regions
Muhammad Kashif Saleemi*, Muhammad Zargham Khan, Ahrar Khan, Muhammad Aamer Mehmood1,2, Muhammad Farooq1,3, Sohail Hameed1, Zahoor ul Hassan, Muhammad Rizwan Javed1,2 and Ijaz Javed4
Department of Pathology, University of Agriculture, Faisalabad-38040, Pakistan; 1National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan; 2Department of Bioinformatics & Biotechnology, Government College University, Faisalabad; 3Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan; 4Department of Physiology and Pharmacology, University of Agriculture, Faisalabad-38040, Pakistan *Corresponding author:


Black Aspergilli are second most important cause of contamination of poultry feed and their by-products with ochratoxin-A after Aspergillus ochraceous in Pakistan. A total of 23 black Aspergilli were isolated from poultry feeds. Of these black Aspergilli samples, 12 (52.17%) were toxigenic (Ochratoxin A). A PCR based molecular method of cloning and sequencing of ITS (internal transcribed spacer) regions of fungal DNA was developed to identify these fungi.  A universal pair of primers (ITS1 and ITS4) was used and PCR product (600 bp) for Aspergillus niger was amplified. These specific universal primers of ITS region were tested for twelve DNA samples of different isolates of Aspergillus species isolated from poultry feeds. The sequencing of ITS-region provided a powerful and reliable tool for identification of black Aspergilli responsible for the production of ochratoxin-A in poultry feed.

Key words: Aspergillus niger, DNA, ITS region, Ochratoxin A, PCR


ISSN 0253-8318 (Print)
ISSN 2074-7764 (Online)