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The Seasonal Toxicological Profile of Four Puffer Fish Species Collected Along Bengal Coast, India

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The Seasonal Toxicological Profile of Four Puffer Fish Species Collected Along Bengal Coast, India Indian Journal of Marine Sciences Vol 33(3), September 2004, pp. 276-280 The seasonal toxicological profile of four puffer fish species collected along Bengal coast, India Somiranjan Ghosh, Alok K. Hazra, Shivaji Banerjee & Biswapati Mukherjee* S. N. Pradhan Centre for Neurosciences, University of Calcutta, 244B, Acharya J. C. Bose Road, Calcutta – 700 020, India *[E-mail : [email protected]] Received 17 November 2003, revised 29 June 2004 Toxicological profiles of the livers and ovaries of four Indian marine puffer fish species viz. Chelonodon patoca, Takifugu oblongus, Lagocephalus lunaris, Lagocephalus inermis collected along coastal Bengal of Digha-Talsari region were evaluated by mouse bioassay. Toxicity expressed in MU (mouse unit)/g in ovaries of all four species were high in monsoon (13.9 – 80 MU/g) and postmonsoon (8.9 – 136 MU/g) seasons during annual reproductive cycle. However, toxicity of livers was much lower (3.2 – 18.5 MU/g) in all the species with seasonal variation. Chelonodon patoca specimens were found to be most toxic and Lagocephalus inermis were least toxic in comparison to other species under investigation. Therefore, health hazard due to puffer fish consumption could be minimised by the information given in this study regarding lethality. [Key words : Puffer, toxicity , Chelonodon patoca, Takifugu oblongus, Lagocephalus inermis, Lagocephalus lunaris, fish] [IPC Code: Int. Cl.7 G01N 33/12] Introduction (Class:Actinopterygii, Order:Tetraodontiformes, Evidence of puffer fish intoxication has been reported Family:Lagocephalidae), Lagocephalus inermis in different geographical locales viz., Japan1, USA2, Temminck & Schlegel (Class:Actinopterygii, Order: Taiwan3, Mexico4, Malaysia5, Bangladesh6 and also in Tetraodontiformes, Family:Lagocephalidae) were India7. Jones8 reported deaths due to ingestion of collected regularly from Digha-Talsari region of cooked roe of puffer Chelonodon patoca for the first Bengal, eastern coast of India (latitude 21°40′N, time in India in 1956. Although the occurrence of longitude 87°30′E) during April 2000 – March 2001. puffer fish poisoning is sporadic in India, there is no The voucher specimens of different puffer fishes have systematic documentation of their toxicological been submitted to National Marine Repository, profiles. The toxicological profiles of wild puffer National Institute of Oceanography, Goa, India. The fishes differ widely amongst different species9, time catch was segregated into three prominent seasons, of collection10 and the geographical region of i.e. premonsoon (March–June, summer; temp 29- 11 occurrence . Even the anatomical distribution of 35oC, rainfall 0.7-19 cm), monsoon (July–October, 12 toxicity varies from species to species . The present rainy; temp 28-33oC, rainfall 8.2-10.6 cm) and work was undertaken to find out the toxicological postmonsoon (November–February, winter; temp 21- profiles of puffer fishes with seasonal variation 28oC, rainfall 0.8-5.8 cm). The puffer species collected along Bengal coast, India and to assess the collected from the coastal waters were immediately toxicity. transferred on ice to the laboratory. After random sampling ten fish specimens of each species were Materials and Methods chosen for toxicity assessment. Livers and ovaries of Four species of marine puffers, viz. Chelonodon each different puffer species, according to the patoca Hamilton (Class:Actinopterygii, Order: respective seasons were dissected out and kept in Tetraodontiformes, Family:Tetraodontidae), Takifugu vacuum-sealed containers below -20°C. oblongus Bloch (Class:Actinopterygii, Order: The liver (10g) and ovary (10g) of each specimen Tetraodontiformes, Family:Tetraodontidae), of different puffer fish species were separately Lagocephalus lunaris Bloch & Schneider homogenized for 5 min with 3 volumes of 1% acetic GHOSH et al : TOXICOLOGICAL PROFILE OF PUFFER FISH 277 acid in methanol and centrifuged (3000 × g) for 15 postmonsoon seasons were 9.5±3.7, 15.0±3.9, min. The residue was extracted twice in the same 18.5±9.3 MU/g respectively indicating slight manner. The supernatants were combined, increment in toxicity through seasons. Matured concentrated under reduced pressure, the residue ovaries were not found in premonsoon season and the dissolved in 0.1% aqueous acetic acid and defatted 3 toxicity was less than 4 MU/g. However, during times with dichloromethane. The aqueous layer was monsoon, fish species with matured ovaries were ultrafiltered with Amicon YM-1 membrane to cut off plenty as evidenced from GI (9.72±2.9) and toxicity substances of more than 1000 daltons. The filtrate was was 80.2 ± 12.9 MU/g. The toxicity of ovaries in freeze-dried and examined for toxicity by mouse postmonsoon season was even more, 136.4±20.3 bioassay. MU /g. Mouse toxicity assay, similar to the method The body weight and length of T. oblongus fish developed for PSP (paralytic shellfish poisoning) specimen were almost same in all three seasons 14 toxin monitoring , has been applied for toxicity (Table 1B). Female fishes are abundant throughout determination among puffers. The mouse bioassay the year. Liver weight in monsoon season (15.0±3.25) was standardized for tetrodotoxin (TTX) in puffers was higher than premonsoon (10.0±2.90) and after Kawabata13 and was expressed as mouse unit postmonsoon (11.0 ± 3.10), as evidenced from (MU) per gram of material, where one MU is the hepatosomatic index. The gonadosomatic index equivalent amount of TTX (0.22 µg) that kills a increased abruptly in monsoon (7.45±1.8) and mouse of ddY strain weighing 20 g in 30 minutes. postmonsoon (5.91±1.6) compared to the premonsoon The lyophilized extract from each puffer specimen was dissolved in 1ml saline solution for mouse (0.56±0.3). Frequencies of toxicity in T. oblongus toxicity assay. Injections were made intraperitoneally specimens were almost cent per cent. The toxicity in to Swiss albino mice (19-21 g) (obtained from Central livers viz., 4.8±2.8, 12.0±4.2, 9.2±1.7 MU/g in Drug Laboratory, Kolkata). A group of three mice premonsoon, monsoon and postmonsoon seasons were used for each experiment. The lethal potency respectively indicated that it was more toxic in was calculated as the time required for killing the monsoon seasons. Ovaries developed substantial mice and was expressed in mouse unit (MU)13. All the toxicity in monsoon (24.5±4.6) and in postmonsoon experiments were statistically analyzed and expressed (22.0±5.9) compared to premonsoon. in Mean ± SD. Lagocephalus lunaris were found plenty throughout seasons (Table 1C). The body weight Results varied slightly, although the lengths were same in Anatomical distribution of toxicity in the puffer three seasons. Female fishes were abundant as Chelonodon patoca is shown in Table 1A. The size of observed in C. patoca (Table 1A) and T. oblongus all fishes collected was almost same and the body (Table 1B). Hepatosomatic index in premonsoon, weight in postmonsoon season (1121± 110.0 g) was monsoon and postmonsoon were 4.6±1.00, 7.9±1.50, nearly double with respect to other seasons. Female 6.0±1.90 respectively which indicated increased liver fishes were more abundant except in monsoon season size in monsoon season. Mature ovaries were found in when male and female fishes were found in equal monsoon and postmonsoon as evident from proportion. The hepatosomatic inedx (liver weight gonadosomatic index 8.05±2.1, 5.56±2.7 respectively. percent with respect to body weight, HI) increased Frequency of toxicity among specimens were almost gradually from premonsoon (8.0±1.2) to monsoon same (84.37, 85.18, 77.78) throughout the seasons. (10.5±0.8) and postmonsoon (14.0±1.6). Liver toxicity was highest in monsoon 7.2±1.8 MU/g Gonadosomatic index (gonadal weight percent with compared to premonsoon (4.9±1.9) and postmonsoon respect to their body weight, GI) in monsoon (6.8±1.6). Ovaries were toxic in monsoon (23.2±6.9) (9.72±2.9), postmonsoon (6.29±2.6) indicated and postmonsoon (8.9±2.5). abundance of gorged ovaries full of matured eggs Lagocephalus inermis fishes were larger in compared to premonsoon (0.44±0.21). From toxic premonsoon (21.35±3.1) than monsoon (18.83±2.9) ratio and frequency of toxicity it was observed that and postmonsoon (18.74±2.4). Similar trend was almost all the species of C. patoca were toxic. The reflected in their average body weight (Table 1D). toxicity of liver in premonsoon, monsoon and Female specimens were available more than male 278 INDIAN J. MAR. SCI., VOL. 33, No. 3, SEPTEMBER 2004 Table 1 ⎯ Anatomical distribution of toxicity in the puffer fishes collected from Bengal coast during March 2000 – February 2001 (Premonsoon - Mar-June, Monsoon – July-Oct, Postmonsoon – Nov-Feb) a b c Seasons Body Length M/F HI GI Toxic Frequency Toxicity (MU/g) weight(g) (cm) ratio ratio of toxicity (%) Liver Ovary (A) Chelonodon patoca Premonsoon 447.5±58.9 31.22±3.2 3 : 4 8.0±1.2 0.44±0.21 18/19 94.73 9.5±3.7 e ND f (n=19d) Monsoon 516.0±78.2 25.59±2.9 1 : 1 10.5±0.8 9.72±2.9 15/15 100.00 15.0±3.9 e 80.2±12.9 e (n= 15 d) Postmonsoon 1121.5±110.0 36.14±4.6 1 : 9 14.0±1.6 6.29±2.6 16/16 100.00 18.5±9.3 e 136.4±20.3 e (n=16 d) (B) Takifugu oblongus Premonsoon 354.0±60.0 26.03±3.1 1 : 4 10.0±2.90 0.56±0.3 14/15 93.34 4.8±2.8 e ND f (n=15d) Monsoon 362.0±72.2 25.46±2.5 1 : 9 15.0±3.25 7.45±1.8 13/13 100.00 12.0±4.2 e 24.5±4.6 e (n= 13 d) Postmonsoon 423.9±48.9 26.25±4.2 3 : 5 11.0±3.10 5.91±1.6 16/16 100.00 9.2±1.7 e 22.0±5.9 e (n=16 d) (C) Lagocephalus lunaris Premonsoon 142.3±13.4 21.70±4.2 2 : 3 4.6±1.00 1.46±0.7 11/13
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