Molecular characterization of ochratoxin a producing indigenous Aspergillus strains from poultry feed in Pakistan
Ochratoxin A (OTA) is nephrocarcinogenic and immunosuppressive toxin and OTA producing molds contaminate the food crops. Isolation and identification of ochratoxin producing fungi was carried out from poultry feed samples (n=120) followed by preliminary confirmation through macroscopic and microscop...
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Universiti Kebangsaan Malaysia
2023
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my-ukm.journal.219172023-07-26T01:34:42Z http://journalarticle.ukm.my/21917/ Molecular characterization of ochratoxin a producing indigenous Aspergillus strains from poultry feed in Pakistan Naz, Gull Anjum, Aftab Ahmad Ali, Tehreem Nawaz, Muhammad Iqbal, Sanaullah Abbas, Mateen Manzoor, Rabia Ochratoxin A (OTA) is nephrocarcinogenic and immunosuppressive toxin and OTA producing molds contaminate the food crops. Isolation and identification of ochratoxin producing fungi was carried out from poultry feed samples (n=120) followed by preliminary confirmation through macroscopic and microscopic characteristics. Purified fungal isolates identified as Aspergillus 1842(91.68%) followed by Penicillium 91 (4.53%), Mucor 52 (2.58), Alternaria 7 (0.35%), Cladosporium 6 (0.29%), Fusarium 4 (0.199%) and unidentified (07). OTA production was confirmed through thin layer chromatography (TLC) followed by high performance liquid chromatography (HPLC). Only 41 isolates (2.22%) out of 1842 Aspergillus isolates were able to produce toxin. At genetic level, characterization was performed through polymerase chain reaction (PCR) using species specific gene primers. From 41 isolates 27, 9 and 5 were characterized as Aspergillus terreus, Aspergillus parasiticus, and Aspergillus ochraceus, respectively. Physical and chemical factors were optimized for OTA production. Under the effect of 37 °C temperature and 7.5 pH of Sabouraud dextrose broth (SDB) medium, higher toxin (969.45±.03 μg/mL) production was observed from ASPO-6 isolate. ASPO-4 isolate produce higher toxin amount in SDB medium with supplementation of maize 5%, wheat 1% and rice 3%. OTA stability was determined by adjusting standard concentration of 100 μg/mL in organic solvents (chloroform, acetonitrile and methanol) and organic solids. Least percentage log reduction in OTA concentration and stability of OTA was observed in opaque vials with chloroform and sucrose and transparent vials with sucrose after 6 months. OTA can be used as indigenous standard for identification of OTA from field samples. Universiti Kebangsaan Malaysia 2023 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/21917/1/ST%2011.pdf Naz, Gull and Anjum, Aftab Ahmad and Ali, Tehreem and Nawaz, Muhammad and Iqbal, Sanaullah and Abbas, Mateen and Manzoor, Rabia (2023) Molecular characterization of ochratoxin a producing indigenous Aspergillus strains from poultry feed in Pakistan. Sains Malaysiana, 52 (3). pp. 821-835. ISSN 0126-6039 http://www.ukm.my/jsm/index.html |
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Ochratoxin A (OTA) is nephrocarcinogenic and immunosuppressive toxin and OTA producing molds contaminate the food crops. Isolation and identification of ochratoxin producing fungi was carried out from poultry feed samples (n=120) followed by preliminary confirmation through macroscopic and microscopic characteristics. Purified fungal isolates identified as Aspergillus 1842(91.68%) followed by Penicillium 91 (4.53%), Mucor 52 (2.58), Alternaria 7 (0.35%), Cladosporium 6 (0.29%), Fusarium 4 (0.199%) and unidentified (07). OTA production was confirmed through thin layer chromatography (TLC) followed by high performance liquid chromatography (HPLC). Only 41 isolates (2.22%) out of 1842 Aspergillus isolates were able to produce toxin. At genetic level, characterization was performed through polymerase chain reaction (PCR) using species specific gene primers. From 41 isolates 27, 9 and 5 were characterized as Aspergillus terreus, Aspergillus parasiticus, and Aspergillus ochraceus, respectively. Physical and chemical factors were optimized for OTA production. Under the effect of 37 °C temperature and 7.5 pH of Sabouraud dextrose broth (SDB) medium, higher toxin (969.45±.03 μg/mL) production was observed from ASPO-6 isolate. ASPO-4 isolate produce higher toxin amount in SDB medium with supplementation of maize 5%, wheat 1% and rice 3%. OTA stability was determined by adjusting standard concentration of 100 μg/mL in organic solvents (chloroform, acetonitrile and methanol) and organic solids. Least percentage log reduction in OTA concentration and stability of OTA was observed in opaque vials with chloroform and sucrose and transparent vials with sucrose after 6 months. OTA can be used as indigenous standard for identification of OTA from field samples. |
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Naz, Gull Anjum, Aftab Ahmad Ali, Tehreem Nawaz, Muhammad Iqbal, Sanaullah Abbas, Mateen Manzoor, Rabia |
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Naz, Gull Anjum, Aftab Ahmad Ali, Tehreem Nawaz, Muhammad Iqbal, Sanaullah Abbas, Mateen Manzoor, Rabia Molecular characterization of ochratoxin a producing indigenous Aspergillus strains from poultry feed in Pakistan |
author_facet |
Naz, Gull Anjum, Aftab Ahmad Ali, Tehreem Nawaz, Muhammad Iqbal, Sanaullah Abbas, Mateen Manzoor, Rabia |
author_sort |
Naz, Gull |
title |
Molecular characterization of ochratoxin a producing indigenous Aspergillus strains from poultry feed in Pakistan |
title_short |
Molecular characterization of ochratoxin a producing indigenous Aspergillus strains from poultry feed in Pakistan |
title_full |
Molecular characterization of ochratoxin a producing indigenous Aspergillus strains from poultry feed in Pakistan |
title_fullStr |
Molecular characterization of ochratoxin a producing indigenous Aspergillus strains from poultry feed in Pakistan |
title_full_unstemmed |
Molecular characterization of ochratoxin a producing indigenous Aspergillus strains from poultry feed in Pakistan |
title_sort |
molecular characterization of ochratoxin a producing indigenous aspergillus strains from poultry feed in pakistan |
publisher |
Universiti Kebangsaan Malaysia |
publishDate |
2023 |
url |
http://journalarticle.ukm.my/21917/1/ST%2011.pdf http://journalarticle.ukm.my/21917/ http://www.ukm.my/jsm/index.html |
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