[Frontiers in Bioscience E3, 74-80, January 1, 2011]

Detection of airborne trichothecene-producing Fusarium species in chicken houses

Yaling Wang1, 2, 3, Tongjie Chai2, Guozhong Lu3, Lijun Sun1, Yi Ouyang3, Xiaodong Sun3

1College of Food Science Technology, Guangdong Ocean University, Jiefang East Road No.40, Xiashan Zone, Zhanjiang 524088, Guangdong Province, P. R. China, 2College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Daizong Street No. 61, Taian 271018, Shandong Province, P. R. China,3College of Life Science, Dalian Nationalities University, Liaohe West Road No. 18, Economic and Technical Development Zone, Dalian 116600, Liaoning Province, P. R. China

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and methods
3.1. Sample Collection
3.2. Fungal cultures and identification
3.3. DNA extraction
3.4. Tri5 gene amplification
3.5. Nucleotide sequencing
3.6. Mycotoxin-producing culture of Tri5 positive isolates
3.7. DON and T-2 detection
4. Results
4.1. Airborne Fusarium isolates
4.2. Tri5-PCR assay
4.3. Detection of mycotoxin related to Tri5 gene
5. Discussion
6. Conclusion
7. Acknowledgements
8. References

1. ABSTRACT

One hundred and forty three airborne Fusarium isolates in chicken houses belonging to seven Fusarium species were analyzed by PCR with Tri5 gene as a specific marker of mycotoxin product . The result of Tri5 gene sequence analysis indicates that the PCR amplification products were 89%-96% identical to the previously reported Tri5 genes, which were all amplified from four F. poae isolates. T-2 toxin and DON was measured by immunoaffinity column and high performance liquid chromatography in Tri5-positive F. poae isolates after being cultured at constant and alternating temperatures. The production of T-2 toxin under alternating temperatures was 14 and 53 times higher than those at constant temperature of 8°C and 25°C. No DON was detected under either testing temperature condition. It is concluded that T-2 toxin-producing F. poae isolates were present in poultry houses, and the concentration of T-2 toxin produced by Tri5-positive F. poae isolates was increased under alternating temperatures. The application of Tri5-PCR associated with IMC-HPLC is an effective and accurate method for rapid detection of T-2 and DON mycotoxins.