Molecular Analysis of the Cyanobacterial Community in Gastric Contents of Egrets with Symptoms of Steatitis

Tomoyasu Nishizawa1, *, Yasuko Neagari2, Takamasa Miura1, , Munehiko Asayama1, Koichi Murata3, Ken-Ichi Harada4, Makoto Shirai1,
1 College of Agriculture, Ibaraki University, Ibaraki 300-0393, Japan
2 Laboratory for Intellectual Fundamentals for Environmental Studies, National Institute for Environmental Studies, Ibaraki 305-8506, Japan
3 College of Bioresource Sciences, Nihon University, Kanagawa 252-0880, Japan;
4 Graduate School of Environmental and Human Science and Faculty of Pharmacy, Meijo University, Aichi 468-8503, Japan;

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© Nishizawa et al.; Licensee Bentham Open.

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* Address correspondence to this author at the College of Agriculture, Ibaraki University, 3-21-1 Chuou, Ami, Ibaraki 300-0393, Japan; Tel: +81 29 888 8687; Fax: +81 29 888 8525; E-mail: Present address: Biological Resource Center, National Institute of Technology and Evaluation, Tokyo 151-0066, Japan; ‡Present address: College of Human and Cultural Sciences, Aikoku Gakuen University, Chiba, 284-0005, Japan


Many deaths of wild birds that have drunk water contaminated with hepatotoxic microcystin-producing cyanobacteria have been reported. A mass death of egrets and herons with steatitis were found at the agricultural reservoir occurring cyanobacterial waterblooms. This study aimed to verify a hypothesis that the egrets and herons which died in the reservoir drink microcystin-producing cyanobacteria and microcystin involves in the cause of death as well as the symptoms of steatitis. The cyanobacterial community in gastric contents of egrets and herons that died from steatitis was assessed using cyanobacterial 16S rRNA-based terminal-restriction fragment length polymorphism (T-RFLP) profiling and a cyanobacterial 16S rRNA-based clone library analysis. In addition, PCR amplification of the mcyB–C region and the mcyG gene, involved in microcystin biosynthesis, was examined. The cyanobacterial community in the gastric contents of two birds showed a simplistic composition. A comparison of cyanobacterial T-RFLP profiling and cloned sequences suggested that the genus Microcystis predominated in both samples of egrets died. Although we confirmed that two egrets which died in the reservoir have taken in cyanobacterial waterblooms containing the genus Microcystis, no mcy gene was detected in both samples according to the mcy gene-based PCR analysis. This study is the first to show the profiling and traceability of a cyanobacterial community in the gastric contents of wild birds by molecular analysis. Additionally, we consider causing symptoms of steatitis in the dead egrets.

Keywords: Agricultural reservoir, cyanobacterial community, gastric content, Microcystis, microcystin biosynthesis (mcy) gene, T-RFLP profiling.