Identification of Causative Ciguatoxins in Red Snappers Lutjanus bohar Implicated in Ciguatera Fish Poisonings in Vietnam
Abstract
:1. Introduction
2. Results
3. Discussion
4. Materials and Methods
4.1. Specimens Collection
4.2. Reagents and Standards
4.3. Genetic Analyses on the Nuclear 16S rDNA and Mit COX1
4.4. Mouse Bioassay
4.5. Clean-Up of Fish Specimens for LC/MS
4.6. Quadrupole LC/MS
4.7. Matrix Effect on the Quadrupole SIM LC/MS for [M+Na]+ of CTX-1B
4.8. Quadrupole Time of Flight (QTOF) LC-MS/MS
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Incident | Implicated Fish | Origin of Implicated Fish | Location of Incident | Date of Incident | Number of Patients/Number of Consumers |
---|---|---|---|---|---|
1 | Red snapper Lutjanus bohar *1 (approx. 5 kg body weight) | Phan Thiet City, Binh Thuan Province | Ho Chi Minh City | 17 July 2014 | 6/6 (family members) |
Symptoms: Developed 3–4 h after eating: Dizziness, headache, pain on muscle and junction joints, nausea, vomiting, and diarrhea for a week. | |||||
2 | Red snapper Lutjanus bohar (approx. 2 kg body weight) | Nha Trang City, Khanh Hoa Province | Nha Trang City, Khanh Hoa Province | 22 June 2016 | 4/4 (family members) |
Symptoms: Developed 1–2 h after eating: Pain in muscles, joints and the back head continuously for several days. After hospitalization for 3 days, the patients came back home but one of them (father) had to be hospitalized again for one more week as symptoms persisted. The father said he felt there was a cold wind blowing into the mouth when drinking water. |
Sample | CTX-1B | 54-deoxy CTX-1B | 52-epi-54-deoxyCTX-1B | |||||||
Ions | [M+NH4]+ | [M+Na]+ | [M+K]+ | [M+NH4]+ | [M+Na]+ | [M+K]+ | [M+NH4]+ | [M+Na]+ | [M+K]+ | |
Formula | C60H90N1O19 | C60H86O19Na | C60H86O19K | C60H90N1O18 | C60H86O18Na | C60H86O18K | C60H90N1O18 | C60H86O18Na | C60H86O18K | |
Theoretical Values | 1128.6102 | 1133.5656 | 1149.5395 | 1112.6152 | 1117.5706 | 1133.5446 | 1112.6152 | 1117.5706 | 1133.5446 | |
Authentic CTX-1B | 1128.6074 (2.4) *1 | 1133.5626 (2.6) | 1149.5345 (4.3) | - | - | - | - | - | - | |
Fish flesh from incident 1 | 1128.6137 (−3.1) | 1133.5688 (−2.9) | 1149.5497 (−8.9) | 1112.6185 (−2.9) | ND | ND | 1112.6091 (5.5) | ND | ND | |
Fish viscera from incident 2 | 1128.6070 (2.8) | ND *2 | ND | 1112.6129 (2.1) | ND | ND | 1112.6073 (7.1) | ND | ND | |
Fish flesh from incident 2 | 1128.6059 (3.8) | 1133.5623 (2.9) | 1149.5359 (3.1) | 1112.6141 (1.0) | ND | ND | 1112.6142 (0.9) | 1117.5734 (−2.5) | ND |
Incident | Body Part | CTX Contents (ng/g) *1 | Total Toxicity (MU/g) Estimated by LC/MS *4 | Toxicity (MU/g) by MBA | ||
---|---|---|---|---|---|---|
CTX-1B Average (SD) | 54-deoxyCTX-1B Average (SD) | 52-epi-54-deoxyCTX-1B Average (SD) | ||||
1 | flesh | 3.67 (0.21) | 0.36 (0.18) *2 | 0.74 (0.24) *2 | 0.58 | 0.15 |
2 | viscera | 0.92 (0.03) | ND | ND *3 | 0.13 | - |
2 | flesh | 2.77 (0.05) | ND | ND *3 | 0.40 | 0.35–0.40 |
Compound | [M+Na]+ m/z | CEP (Volts) |
---|---|---|
CTX-1B(P-CTX-1) | 1133.6 | 42 |
52-epi-54-deoxyCTX-1B (P-CTX-2) | 1117.6 | 41 |
54-deoxyCTX-1B (P-CTX-3) | 1117.6 | 41 |
CTX-4A | 1083.6 | 40 |
CTX-4B | 1083.6 | 40 |
M-seco-CTX-4A/B | 1101.6 | 41 |
CTX-3C | 1045.6 | 39 |
49-epi-CTX-3C | 1045.6 | 39 |
51-hydroxyCTX-3C | 1061.6 | 40 |
2-hydroxyCTX-3C | 1063.6 | 40 |
2,3-dihydroxyCTX-3C | 1079.6 | 40 |
M-seco-CTX-3C | 1063.6 | 40 |
M-seco-CTX-3C methyl acetal | 1077.6 | 40 |
gambierol | 779.5 | 32 |
gambieric acid A | 1101.6 *1 | 41 |
gambieric acid B | 1115.6 *1 | 41 |
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Share and Cite
Ha, D.V.; Uesugi, A.; Uchida, H.; Ky, P.X.; Minh, D.Q.; Watanabe, R.; Matsushima, R.; Oikawa, H.; Nagai, S.; Iwataki, M.; et al. Identification of Causative Ciguatoxins in Red Snappers Lutjanus bohar Implicated in Ciguatera Fish Poisonings in Vietnam. Toxins 2018, 10, 420. https://doi.org/10.3390/toxins10100420
Ha DV, Uesugi A, Uchida H, Ky PX, Minh DQ, Watanabe R, Matsushima R, Oikawa H, Nagai S, Iwataki M, et al. Identification of Causative Ciguatoxins in Red Snappers Lutjanus bohar Implicated in Ciguatera Fish Poisonings in Vietnam. Toxins. 2018; 10(10):420. https://doi.org/10.3390/toxins10100420
Chicago/Turabian StyleHa, Dao Viet, Aya Uesugi, Hajime Uchida, Pham Xuan Ky, Dang Quoc Minh, Ryuichi Watanabe, Ryoji Matsushima, Hiroshi Oikawa, Satoshi Nagai, Mitsunori Iwataki, and et al. 2018. "Identification of Causative Ciguatoxins in Red Snappers Lutjanus bohar Implicated in Ciguatera Fish Poisonings in Vietnam" Toxins 10, no. 10: 420. https://doi.org/10.3390/toxins10100420