Search Results (7)

Ingesting fish contaminated with ciguatoxins (CTXs) originating from epibenthic dinoflagellates causes ciguatera fish poisoning (CFP). CFP occurs mainly in the tropical and subtropical Indo–Pacific region and the Caribbean Sea. Furthermore, it occurs sporadically in Japan, especially in the Ryukyu Islands between Taiwan and Kyushu, Japan. Variola louti is the most frequently implicated fish with a suggested toxin profile, consisting of ciguatoxin-1B and two deoxy congeners. Therefore, using the liquid chromatography–tandem mass spectrometry (LC-MS/MS), we analyzed CTXs in the flesh of 154 individuals from various locations and detected CTXs in 99 specimens (64%). In 65 fish (43%), CTX levels exceeded the Food and Drug Administration (FDA) guidance level (0.01 µg/kg). Furthermore, in four specimens (3%), the guideline level in Japan (>0.18 µg/kg) was met. Additionally, although the highest total CTX level was 0.376 µg/kg, the consumption of 180 g of this specimen was assumed to cause CFP. Moreover, only CTX1B, 52-epi-54-deoxyCTX1B, and 54-deoxyCTX1B were detected, with the relative contribution of the three CTX1B analogs to the total toxin content (35 ± 7.7 (SD)%, 27 ± 8.1%, and 38 ± 5.6%, respectively) being similar to those reported in this region in a decade ago. Subsequently, the consistency of the toxin profile in V. louti was confirmed using many specimens from a wide area. As observed, total CTX levels were correlated with fish sizes, including standard length (r = 0.503, p = 3.08 × 10⁻¹¹), body weight (r = 0.503, p = 3.01 × 10⁻¹¹), and estimated age (r = 0.439, p = 3.81 × 10⁻⁷) of the specimens. Besides, although no correlation was observed between condition factor (CF) and total CTX levels, a significance difference was observed (p = 0.039) between the groups of skinnier and fattier fish, separated by the median CF (3.04). Results also showed that the CF of four specimens with the highest CTX level (>0.18 µg/kg) ranged between 2.49 and 2.87, and they were skinnier than the average (3.03) and median of all specimens. Full article

Ciguatera poisoning (CP) cases linked to the consumption of deep-water fish occurred in 2003 in the Gambier Islands (French Polynesia). In 2004, on the request of two local fishermen, the presence of ciguatoxins (CTXs) was examined in part of their fish catches, i.e., 22 specimens representing five deep-water fish species. Using the radioactive receptor binding assay (rRBA) and mouse bioassay (MBA), significant CTX levels were detected in seven deep-water specimens in Lutjanidae, Serranidae, and Bramidae families. Following additional purification steps on the remaining liposoluble fractions for 13 of these samples (kept at −20 °C), these latter were reanalyzed in 2018 with improved protocols of the neuroblastoma cell-based assay (CBA-N2a) and liquid chromatography tandem mass spectrometry (LC–MS/MS). Using the CBA-N2a, the highest CTX-like content found in a specimen of Eumegistus illustris (Bramidae) was 2.94 ± 0.27 µg CTX1B eq. kg⁻¹. Its toxin profile consisted of 52-epi-54-deoxyCTX1B, CTX1B, and 54-deoxyCTX1B, as assessed by LC–MS/MS. This is the first study demonstrating that deep-water fish are potential ciguatera vectors and highlighting the importance of a systematic monitoring of CTXs in all exploited fish species, especially in ciguatera hotspots, including deep-water fish, which constitute a significant portion of the commercial deep-sea fisheries in many Asian–Pacific countries. Full article

Ciguatoxins (CTXs) are polyether marine biotoxins that can cause ciguatera poisoning (CP) after the consumption of fish or invertebrates containing sub ppb levels; concentrations that present a challenge for current extraction and analysis methods. Here, a newly developed and (partly) validated single-day extraction protocol is presented. First, the fish sample is broken-down by enzymatic digestion, followed by extraction and extract clean-up by defatting and two solid-phase extractions. Final extracts were investigated using two different CTX-analysis methods; an in vitro cytotoxicity assay (N2a-assay) and by LC-MS/MS. Validation was performed for both fillet and freeze-dried samples of snapper, parrotfish, and grouper spiked with CTX1B, 52-epi-54-deoxyCTX1B, 54-deoxyCTX1B, and CTX3C. Based on recovery rates (35–88%) and matrix effects (66–116%) determined by LC-MS/MS, the enzyme protocol is applicable to various matrices. The protocol was applied to naturally contaminated fish tissue (Lutjanus bohar) obtained during a CP incident in Germany. Several potential CTX congeners were identified by a two-tier LC-MS/MS approach (screening of sodium adducts, high-resolution or low-resolution confirmation via ammonium adducts). Inclusion of >30 known CTX congeners into the LC-MS/MS methods and single-day sample preparation make the method suitable for analysis of ciguatera suspect samples at sub ppb levels also with undisclosed CTX profiles. Full article

Ciguatera fish poisoning (CFP) is one of the most frequently encountered seafood poisoning syndromes; it is caused by the consumption of marine finfish contaminated with ciguatoxins (CTXs). The majority of CFP cases result from eating fish flesh, but a traditional belief exists among people that the head and viscera are more toxic and should be avoided. Unlike the viscera, scientific data to support the legendary high toxicity of the head is scarce. We prepared tissue samples from the fillet, head, and eyes taken from five yellow-edged lyretail (Variola louti) individuals sourced from Okinawa, Japan, and analyzed the CTXs by LC-MS/MS. Three CTXs, namely, CTX1B, 52-epi-54-deoxyCTX1B, and 54-deoxyCTX1B, were confirmed in similar proportions. The toxins were distributed nearly evenly in the flesh, prepared separately from the fillet and head. Within the same individual specimen, the flesh in the fillet and the flesh from the head, tested separately, had the same level and composition of toxins. We, therefore, conclude that flesh samples for LC-MS/MS analysis can be taken from any part of the body. However, the tissue surrounding the eyeball displayed CTX levels two to four times higher than those of the flesh. The present study is the first to provide scientific data demonstrating the high toxicity of the eyes. Full article

“Ciguatera” fish poisoning (CFP) is one of the well-known food poisoning caused by the ingestion of fish that have accumulated trace amounts of ciguatoxins (CTXs). CFP affects more than 50,000 individuals annually. The difficulty in preventing CFP comes from the lack of reliable methods for analysis of CTXs in contaminated fish, together with the normal appearance, taste, and smell of CTX-contaminated fish. Thus, a sensitive, accurate, routine, and portable analytical method to detect CTXs is urgently required. Monoclonal antibodies (mAbs) specific against either wing of major CTX congeners (CTX1B, 54-deoxyCTX1B, CTX3C, and 51-hydroxyCTX3C) were generated by immunizing mice with rationally designed synthetic haptens-KLH conjugates instead of the CTXs. Haptenic groups with a surface area greater than 400 Ų are required to produce mAbs that can strongly bind to CTXs. Furthermore, a highly sensitive fluorescence-based sandwich enzyme-linked immunosorbent assay (ELISA) was developed. This protocol can detect and quantify four major CTX congeners (CTX1B, 54-deoxyCTX1B, CTX3C, and 51-hydroxyCTX3C) with a limit of detection (LOD) of less than 1 pg/mL. The LOD determined for this sandwich ELISA is sufficient to detect CTX1B-contaminated fish at the FDA guidance level of 0.01 ppb. Full article

Ciguatera fish poisoning (CFP) is a type of food poisoning caused by the consumption of a variety of toxic ciguatera fish species in the tropical and subtropical waters. Although there have been a large number of suspected CFP cases in the Southeast Asian countries, few were confirmed with causative ciguatoxins (CTXs), and reliable information on the symptoms still remains rather limited. In the present study, CTXs in red snapper Lutjanus bohar, implicated in two suspected CFP cases in Vietnam in 2014 and 2016, were determined by use of the single-quadrupole selected ion monitoring (SIM) liquid chromatography/mass spectrometry (LC/MS). Ciguatoxin-1B (CTX-1B), 54-deoxyCTX-1B, and 52-epi-54-deoxyCTX-1B were detected in the red snapper by our LC/MS method. Moreover, CTX-1B, 54-deoxyCTX-1B, and 52-epi-54-deoxyCTX-1B were further identified by the time of flight (TOF) LC/MS with the exact mass spectrum. The CTX profile of the red snapper in Vietnam is similar to those of ciguatera fish from Australia, Okinawa Islands in Japan, Kiribati, and Hong Kong. This is the first comprehensive report unambiguously identifying the causative toxins in fish implicated with reliable information on the poisoning symptoms in CFP in Vietnam and/or Southeast Asian countries. Full article

Ciguatoxins (CTXs) contaminate fish worldwide and cause the foodborne illness ciguatera. In the Pacific, these toxins are produced by the dinoflagellate Gambierdiscus toxicus, which accumulates in fish through the food chain and undergoes oxidative modification, giving rise to numerous analogs. In this study, we examined the oxidation of CTXs in vitro with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis using reference toxins, and found that CTX4A, CTX4B, and CTX3C, which are produced by the alga, are oxidized to the analogs found in fish, namely CTX1B, 52-epi-54-deoxyCTX1B, 54-deoxyCTX1B, 2-hydroxyCTX3C, and 2,3-dihydroxyCTX3C. This oxidation was catalyzed by human CYP3A4, fish liver S9 fractions, and microsomal fractions prepared from representative ciguateric fishes (Lutjanus bohar, L. monostigumus, and Oplegnathus punctatus). In addition, fish liver S9 fractions prepared from non-ciguateric fishes (L. gibbus and L. fulviflamma) in Okinawa also converted CTX4A and CTX4B to CTX1B, 54-deoxyCTX1B, and 52-epi-54-deoxyCTX1B in vitro. This is the first study to demonstrate the enzymatic oxidation of these toxins, and provides insight into the mechanism underlying the development of species-specific toxin profiles and the fate of these toxins in humans and fish. Full article