Toxicity of puffer fish fins

Puffer fish is prized as a Japanese traditional food and its fin is also used in the cuisine. However, whether the fin is edible or not is determined for convenience from the toxicity of skin, since little information is available about the toxicity of puffer fish fins. In the present study, we examined the toxicity of fins and skin of three toxic species, Takifugu vermicularis, T. snyderi, and T. porphyreus. The toxicity of T. vermicularis fins (< 5-52.4 MU/g) was significantly lower than that of skin (<5-1200 MU/g). HPLC analysis showed that tetrodotoxin was a major toxic principle irrespective of the toxicity value in each tissue of T. vermicularis. In the case of T. snyderi and T. porphyreus, the toxicity of fins was at almost the same level as that of the skin. The toxicity (< 10-12 MU/g) of caudal fins of T. porphyreus was apparently increased to 16.5-22.0 MU/g by drying. However, the toxin amounts in the dried fins were slightly decreased as compared with those of the non-dried fins. These results demonstrate that puffer fish with toxic skin also have toxic fins.     

Tetrodotoxin poisoning due to smooth-backed blowfish Lagocephalus inermis and toxicity of L. inermis caught off the Kyushu coast, Japan

Food poisoning due to ingestion of a puffer fish occurred in Nagasaki Prefecture, Japan, in October 2008, causing neurotoxic symptoms similar to those of tetrodotoxin (TTX) poisoning. In the present study, we identified the species, toxicity, and toxins using the remaining samples of the causative puffer fish. The puffer fish was identified as smooth-backed blowfish Lagocephalus inermis by nucleotide sequence analysis of the 16S rRNA and cytochrome b gene fragments of muscle mitochondrial DNA. The residual liver sample showed toxicity as high as 1,230 mouse unit (MU)/g by bioassay and TTX was detected by liquid chromatography/mass spectrometry analysis. We therefore concluded that the food poisoning was due to TTX caused by consumption of the toxic liver of L. inermis. This is the first report that the liver of L. inermis caught in Japanese waters is strongly toxic, with levels exceeding 1,000 MU/g. In this context, we re-examined the toxicity of L. inermis collected off the coast of Japan. Of 13 specimens assayed, 12 were toxic, although the toxicity varied markedly among individuals and tissues. Because the intestine and ovary of L. inermis have been considered non-toxic, it is particularly noteworthy that these organs were determined to be toxic, with a maximum toxicity of 43.6 MU/g and 10.0 MU/g, respectively. Furthermore, kidney, gallbladder, and spleen, whose toxicity has been unknown, were frequently found to be weakly toxic with levels ranging from 10 to 99 MU/g. Therefore, further study is needed to re-examine the toxicity of smooth-backed blowfish L. inermis in the coastal waters of Japan.     

Toxicity of puffer fish cultured in netcages

During 1990 to 2003, the toxicity of the liver in 4,515 specimens of the puffer fish Takifugu rubripes (torafugu) cultured in netcages or on land were investigated by means of mouse bioassay and liquid chromatography-mass spectrometry (LC/MS). Other tissues (skin, muscles, gonads, etc.) were also investigated in some of them. All the livers and other parts examined were found to be non-toxic. The peak corresponding to tetrodotoxin (TTX) was not detected in the samples by LC/MS analysis for TTX (< 0.1 MU/g). These results show that puffer fish fed on a non-toxic diet in netcages do not become intoxicated.     

Identification of tetrodotoxin and fish species in a dried dressed fish fillet implicated in food poisoning

There were five victims of neurotoxic food poisoning from a dried dressed fish fillet in Changhua County, Taiwan, in February 2000. The toxicity of the dried dressed fish fillets was 243 mouse units per g according to a tetrodotoxin bioassay. The partially purified toxin was identified as tetrodotoxin and anhydrotetrodotoxin. The sequence of the 376-nucleotide region in the cytochrome b gene of the mitochondrial DNA exhibited the same genotype as that of the toxic puffer fish Lagocephalus lunaris. The same single restriction site for Hinfl was found in the polymerase chain reaction (PCR) products from the dried dressed fish fillet and the muscle of L. lunaris, yielding two DNA fragments of 170 and 206 bp. However, no restriction site for Hinfl was found in the PCR products from other toxic puffer fishes, including Takifugu niphobles, Takifugu oblongus, and Takifugu rubripes. Therefore, the species of the dried dressed fish fillet was identified as L. lunaris and its causative agent was identified as tetrodotoxin.     

不同烹饪方式对盐渍鱼干风味和微观结构的影响

本文对比研究了微波烹饪、空气炸锅烹饪、烘烤烹饪和蒸煮烹饪4种常用烹饪方式对盐渍鱼干咀嚼性(质构)、滋味(游离氨基酸),风味(挥发性物质)和微观结构的影响。结果表明:4种烹饪方式均可通过显著降低盐渍鱼干的硬度和咀嚼性、增加弹性来改变其质构特性;微波烹饪、空气炸锅烹饪和烤箱烹饪都能不同程度的降低盐渍鱼干的游离氨基酸总量,而蒸煮烹饪提高了氨基酸总量,从原始量586.66 mg/100 g增加至631.45 mg/100 g;4种烹饪方式均可使鱼干样品中醛酮类物质和酯类物质含量显著上升,烷烃类物质含量显著下降,其中空气炸锅烹饪处理鱼干样品中醛酮类物质含量最高,达到37.07%;烹饪处理使盐渍鱼干肌肉纤维严重皱缩,纤维束间充满了烹饪用油,空气炸锅和烘烤烹饪处理后盐渍鱼干纤维束间的空隙较明显。     

Assessment of Enterotoxin Production and Cross‐Contamination of Staphylococcus aureus between Food Processing Materials and Ready‐To‐Eat Cooked Fish Paste

This study evaluated Staphylococcus aureus growth and subsequent staphylococcal enterotoxin A production in tryptone soy broth and on ready‐to‐eat cooked fish paste at 12 to 37 °C, as well as cross‐contamination between stainless steel, polyethylene, and latex glove at room temperature. A model was developed using Barany and Roberts's growth model, which satisfactorily described the suitable growth of S. aureus with R²‐adj from 0.94 to 0.99. Except at 12 °C, S. aureus cells in TSB presented a lag time lower (14.64 to 1.65 h), grew faster (0.08 to 0.31 log CFU/h) and produced SEA at lower cell density levels (5.65 to 6.44 log CFU/mL) compare to those inoculated on cooked fish paste with data of 16.920 to 1.985 h, 0.02 to 0.23 log CFU/h, and 6.19 to 7.11 log CFU/g, respectively. Staphylococcal enterotoxin type A (SEA) visual immunoassay test showed that primary SEA detection varied considerably among different storage temperature degrees and media. For example, it occurred only during exponential phase at 30 and 37 °C in TSB, but in cooked fish paste it took place at late exponential phase of S. aureus growth at 20 and 25 °C. The SEA detection test was negative on presence of S. aureus on cooked fish paste stored at 12 and 15°C, although cell density reached level of 6.12 log CFU/g at 15 °C. Cross‐contamination expressed as transfer rate of S. aureus from polyethylene surface to cooked fish paste surface was slower than that observed with steel surface to cooked fish paste under same conditions. These results provide helpful information for controlling S. aureus growth, SEA production and cross‐contamination during processing of cooked fish paste.     

Transfer profile of orally and intramuscularly administered tetrodotoxin to artificial hybrid specimens of the pufferfish Takifugu rubripes and Takifugu porphyreus

Tetrodotoxin (TTX) was administered to artificially hybridized specimens of the pufferfish Takifugu rubripes and Takifugu porphyreus to investigate toxin accumulation in hybrids and TTX transfer/accumulation profiles in the pufferfish body. In test fish administered TTX-containing feed homogenate at a dose of ～400 MU/fish by oral gavage using a syringe (OGA group), the toxin content (MU/g tissue) of the digestive tract rapidly decreased and that of the liver increased from 1 to 24 h after administration. From 24 to 120 h, the toxin content of the liver decreased gradually, and the toxin appeared in the skin. On the other hand, intramuscularly administered TTX (400 MU/fish) was rapidly transferred to the liver and skin via the blood, and only a little toxin remained in the muscle even at 1 h (IMA group). The total amount of toxin remaining in the whole body (% of administered toxin) was 31-45% in the OGA group, and 42-74% in the IMA group; the scores in the OGA group were generally lower than those in the IMA group. In both OGA and IMA groups, the greatest amount of toxin accumulated in the liver (23-52%) after 8 h, followed by the skin (11-21%) after 72 h. The TTX administration experiment, especially using the oral gavage administration method, revealed that skins and livers of 'torama' pufferfish hybrid are endowed with TTX-accumulating ability, but the muscles are not, and that TTX taken up from toxic feed to the pufferfish body is transferred first to the liver and then to the skin via the blood.     

Sterol Content of Fish,Crustacea and Mollusc: Effects of Cooking Methods

Sterol contents (desmosterol, cholesterol, stigmasterol, and sitosterol) of 36 fresh and cooked seafood and four freshwater fish species were investigated by high performance liquid chromatography (HPLC) method. Cholesterol and sitosterol were main sterols in seafood and freshwater fish species. Raw shellfish and molluscs contained cholesterol above 18.92 mg/100 g fish muscle, while the cholesterol content of marine fish ranged from 6.5 to 78.40 mg/100 g fish muscle. The cooking process resulted in significant effects on the sterol contents of seafood and freshwater species (p < 0.05). A remarkable increase in sitosterol (more than 3–4 fold compared to raw fish) was recorded for some fish species cooked in the oven. The highest desmosterol content was observed for fried fish, whereas the frying process resulted in a significant loss in cholesterol and sitosterol contents of marine fish (p < 0.05). The impacts of cooking methods on sterol content of seafood and their form varied depending on fish species and the cooking method used.     

Quantitative Risk Assessment for Formalin Treatment in Fish Preservation: Food Safety Concern in Local Market of Bangladesh

In Bangladesh, fishes are adulterated by hazardous chemicals at different steps from farm to consumers. Formalin (FA) is reported to be frequently added as preservative either by dipping or spraying to the fresh fishes by the fish traders while transporting to domestic marketing chain to prevent spoilage and extend shelf life. Thus, the objective of the present study was toconduct quantitative risk assessment (QRA) for formalin treated fish in Bangladesh. The probabilistic QRA of formalin treated fish was performed based on available secondary data. Available data on concentration of formalin in fish, daily fish consumption by the consumer and their body weight were used to estimate the risk of residual formalin to the consumers. Based on the data, three different scenarios (average consumption, two and four times of average consumption considered as scenarios 1, 2 and 3, respectively) were used for exposure analysis using @Risk program version 6.0.FA concentration in consumedfresh and cooked (boiling) fish was 5.34x10^-02 and 2.340x10^-02 (mg/kg bw/day), respectively and national average fish consumption was 200 g/day. QRA reveals that FA intake under scenario1 and 2 was lower than acceptable daily intake (ADI 0.2 mg/kg bw/day set by the United States Environmental Protection Agency) thus lower risk observed for both fresh and cooked fish. However, scenario 3 revealed that 0.01% population was at risk (FA intake 0.21 mg/kg bw/day higher than ADI) upon the fresh fish consumption, where cooked fish (FA 9.38x10^-02 mg/kg bw/day) consumer remains safe at the same scenario. The result confirmed that cooking has significant effect to reduction of formalin. Therefore, probabilistic quantitative risk assessment of formalin treated fish could provide important risk information to the risk manager (government), whether the population is at risk or not? The result could be applied to establish effective risk management strategy in Bangladesh.     

Taster Response to Salt in Minced Fish

Cooked fish minces, alone and with four added levels of salt, were tasted at the same sessions as five solutions of similar salt concentration. Saltiness was measured by the tasters using the method of magnitude estimation. The magnitude estimation exponent for salt in cooked fish mince was 1.05 whereas that for the solution was significantly higher at 1.51.