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. 2020 Oct;31(3):91-107.
doi: 10.21315/tlsr2020.31.3.7. Epub 2020 Oct 15.

Fumonisin B1 Production by Fusarium Species and Mycotoxigenic Effect on Larval Zebrafish

Najihah Azman  1 , Nur Ain Izzati Mohd Zainudin  1 , Wan Norhamidah Wan Ibrahim  1
Affiliations

Fumonisin B1 Production by Fusarium Species and Mycotoxigenic Effect on Larval Zebrafish

Najihah Azman et al. Trop Life Sci Res. 2020 Oct.

Abstract
in English, Malay (macrolanguage)

Fumonisin B1 (FB1) is a common mycotoxin produced by Fusarium species particularly F. proliferatum and F. verticillioides. The toxin produced can cause adverse effects on humans and animals. The objectives of this study were to detect the production of FB1 based on the amplification of FUM1 gene, to quantify FB1 produced by the isolates using Ultra-fast Liquid Chromatography (UFLC) analysis, to examine the embryotoxicity effect of FB1 and to determine EC50 toward the larvae of zebrafish (Danio rerio). Fifty isolates of Fusarium species were isolated from different hosts throughout Malaysia. Successful amplification of the FUM1 gene showed the presence of this gene (800 bp) in the genome of 48 out of 50 isolates. The highest level of FB1 produced by F. proliferatum isolate B2433 was 6677.32 ppm meanwhile F. verticillioides isolate J1363 was 954.01 ppm. From the assessment of embryotoxicity test of FB1 on larvae of zebrafish, five concentrations of FB1 (0.43 ppm, 0.58 ppm, 0.72 ppm, 0.87 ppm and 1.00 ppm) were tested. Morphological changes of the FB1 exposed-larvae were observed at 24 to 168 hpf. The mortality rate and abnormality of zebrafish larvae were significantly increased at 144 hpf exposure. Meanwhile, the spontaneous tail coiling showed a significant difference. There were no significant differences in the heartbeat rate. As a conclusion, the presence of FUM1 in every isolate can be detected by FUM1 gene analysis and both of the species produced different concentrations of FB1. This is the first report of FB1 produced by Fusarium species gave a significant effect on zebrafish development.

Fumonisin B1 (FB1) ialah mikotoksin biasa yang dihasilkan oleh spesies Fusarium, terutamanya F. proliferatum and F. verticillioides. Toksin yang dihasilkan boleh menyebabkan kesan yang buruk terhadap manusia dan haiwan. Objektif kajian ini adalah untuk mengesan penghasilan FB1 berdasarkan amplifikasi gen FUM1, mengukur kuantiti FB1 yang dihasilkan oleh pencilan menggunakan analisis Kromatografi Cecair Ultra-Cepat (UFLC), memeriksa kesan embriotoksik FB1 dan mengenal pasti EC50 terhadap ikan zebra (Danio rerio). Lima puluh spesies Fusarium diasingkan daripada perumah berlainan di seluruh Malaysia. Amplifikasi gen FUM1 pada 48 daripada 50 pencilan berjaya menunjukkan kehadiran gen ini (800 bp). Pencilan F. proliferatum B2433 menghasilkan FB1 pada aras tertinggi sebanyak 6677.32 bpm manakala pencilan F. verticillioides J1363 sebanyak 954.01 ppm. Berdasarkan penilaian ujian embriotoksik FB1 ke atas larva ikan zebra, lima kepekatan FB1 (0.43 ppm, 0.58 ppm, 0.72 ppm, 0.87 ppm dan 1.00 ppm) telah diuji. Perubahan morfologi larva yang terdedah kepada FB1 diperhatikan pada 24 hingga 168 hpf. Kadar kematian dan ketidaknormalan larva ikan zebra meningkat pada pendedahan 144 hpf. Sementara itu, penggulungan ekor secara spontan menunjukkan perbezaan yang bererti. Tiada perbezaan yang bererti terhadap kadar degupan jantung. Kesimpulannya, kehadiran FUM1 pada setiap pencilan boleh dikesan dengan analisis gen FUM1 dan kedua-dua spesies menghasilkan kepekatan FB1 yang berbeza. Hasil kajian ini merupakan laporan pertama FB1 yang dihasilkan oleh spesies Fusarium yang memberikan kesan yang bererti ke atas perkembangan ikan zebra.

Keywords: Fumonisin B1; Fusarium; Malaysia; Toxic; Ultra-Fast Liquid Chromatography.

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Figures

Figure 1
Figure 1
Representative optical images of deformed zebrafish larvae at 24 hpf caused by FB1 exposure. Malformations are indicated by arrows. BC: blood clotting. Scale bar: 500 μm.
Figure 2
Figure 2
Representative optical images of deformed zebrafish larvae at 48 hpf, 72 hpf and 96 hpf caused by FB1 exposure. Malformations are indicated by arrows. SC: spinal curvature; TM: tail malformation; PE: pericardial edema; YSE: yolk sac edema; BC: blood clotting. Scale bar: 500 μm.
Figure 3
Figure 3
Representative optical images of deformed zebrafish larvae at 120 hpf, 144 hpf and 168 hpf caused by FB1 exposure. Malformations are indicated by arrows. SC: spinal curvature; TM: tail malformation; PE: pericardial edema; YSE: yolk sac edema. Scale bar: 500 μm.
Figure 4
Figure 4
Mortality rate of zebrafish larvae exposed with different concentrations of FB1 according to hours post-fertilisation (hpf).
Figure 5
Figure 5
Concentration-response curve of FB1 towards zebrafish.
See this image and copyright information in PMC

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