Author/Authors :
Saleemi, Muhammad Kashif University of Agriculture - Department of Pathology, Pakistan , Khan, Muhammad Zargham University of Agriculture - Department of Pathology, Pakistan , Khan, Ahrar University of Agriculture - Department of Pathology, PAKISTAN , Mehmood, Muhammad Aamer National Institute for Biotechnology and Genetic Engineering, Pakistan , Mehmood, Muhammad Aamer Government College University - Department of Bioinformatics and Biotechnology, Pakistan , Farooq, Muhammad National Institute for Biotechnology and Genetic Engineering, Pakistan , Farooq, Muhammad Niigata University - Graduate School of Medical and Dental Sciences, Japan , Hameed, Sohail National Institute of Biotechnology and Genetic Engineering, PAKISTAN , Zahoor-ul-Hassan University of Agriculture - Department of Pathology, Pakistan , Javed, Muhammad Rizwan National Institute for Biotechnology and Genetic Engineering, Pakistan , Javed, Muhammad Rizwan Government College University - Department of Bioinformatics and Biotechnology, Pakistan , Javed, Ijaz University of Agriculture - Department of Physiology and Pharmacology, Pakistan
Abstract :
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 A. 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.
Keywords :
Aspergillus niger , DNA , ITS region , Ochratoxin A , PCR
