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Retention of ciguatera toxin by the red snapper, Lutjanus bohar
69
Abstracts ERspAMER, V., RosEGmNi, M., ENDEAN, R. and ANAsTAsi, A. (Institute of Pharmacology, Univ. of Parma, Parma, Italy ; Dept . of Zoology, Univ . of Queensland, Brisbane, Australia; Research Institute, Farmitalia, Milano, Italy). Biogenic amines and active polypeptides in the skin of Australian amphibians . Natitre Lond. 212, 204, 1966 .
EIGHTEEN species of Australian amphibian were examined . Nine to ten of them were shown to contain considerable amounts of 5-hydroxyindolealkylamines, represented not only by 5-hydroxytryptannine, but also by its N-methyl derivatives. Large amounts of histamine were present only in Hyla caerulea and Hyla infrafrenata . Leptodactyline was always lacking. Physalaemin- and bradykinin-like polypeptides were not detectable, but a new, highly active polypeptide, Caerulein, was discovered in the skin of Hyla caerulea and other species of Hyla. Caerulein possesses, in addition to a powerful, relatively long-lasting hypotensive action, conspicuous effects on a number of external secretions as well as on some extravascular smooth muscles. The possible interest of this research from the view point of biochemical taxonomy is emphasized . V.E .
RtcHARDs, G. M., DU VAIR GENEVIEVE, and LAsxowsxr, SR., M. (Biochemical Laboratory for Cancer Research, Marquette University School of Medicine, Milwaukee, Wisconsin, U.S.A.) . Comparison of the levels of phosphodiesterase, endonuclease, and monophosphatases in several snake venoms . Biochemistry 4, 501, 1965
THE conmoN use of venom phosphodiesterase (PDase) in studying nucleotide sequences requires the use of a preparation of PDase free from contaminating enzymes. In this paper the venoms from five species of snakes were analysed for PDase, 5'-nucleotidase (5'-Nase), endonuclease (ENase), and alkaline nonspecific phosphatase (PMase). The authors were able to determine ENase directly in crude venom by the spectrophotometric method using very dilute solutions of the venom. determination of ENase activity in crude venom had not been successful. Thevenoms of Agkistrodon piscivorus, Bothrops atrox, Crotalus adamanteus, and C. atroxclosely resemble each other in contents and relative proportions of the four enzymes. The PMase activity of the venom of Naja nigricollis is unusually high . Since it is extremely difficult to separate PMase from PDase, Naja venom should be ruled out from the sources for the purification of PDase, a fact recently demonstrated by Russell (Toxicon 4, 153 (1966)). A.O . BRAGANCA, B. M. and KHANDEPARYAR, V. G. (BiochemistryDivision, Indian Cancer Research Centre,Bombay, India) . Phospholipase A activity of cobra venom and lysis of Yoshida sarcoma cells. Life Sciences 5, 1911, 1966 .
A compoNENT of cobra venom labelled P, obtained by column chromatography and fractionation with (NH,),SO, appears to have phospholipase C activity . Incubation of P, with phosphatidyl ethanolamine or phosphotidyl serine, or their lyso derivatives and subsequent analysis by thin-layer chromatography indicated the production of O-phosphoryl ethanolamine and O-phosphoryl serine respectively . The formation of diglycerides could not always be detected perhaps due to traces of phospholipase A and B in P, . Lecithin was resistant to the phospholipase C activity of P, . Phosphatidyl ethanolamine and serine present in the Yoshida sarcoma cells were also susceptible to attack by the P, preparation indicating that observed lysis of these cells by P, may be due to its phospholipase C activity . In mice P, had less than 1 per cent of the toxicity of a purified toxin fraction of cobra venom and less than 0*2 per cent of the hemolytic activity of a purified phospholipase A fraction . P.R . BANNER, A. H., HoLFmcH, P. and PrYAKARNCHANA, T. (Hawaii Marine Laboratory, University of Hawaii, Honolulu, Hawaii). Retention of ciguatera toxin by the red snapper, Lutjanus bohar. Copela 2, 297, 1966.
A sAMPLE of 66 samples of the red snapper, Lutjanus bohar, from Christmas Island, Line Islands, whose initial level of toxicity was approximated statistically, was maintained in the holding ponds at the Hawaii Marine Laboratory, Honolulu, for varying periods from 3 to 30 months on a nontoxic diet . At the end of the period the fish showed no significant decline in their toxicity . (Authors' abstract).
70
Abstracts
This work establishes the feasibility of the food chain hypothesis in ciguatera intoxication after the ingestion of fish because it shows effectively that the toxin level in the tissue of fish maintained for long periods of time on a nontoxic diet does not diminish . J.F .G . RorrMAN, M. I. (Science Research Institute Vaccines, Minister Public Health, Taschkent, U.Z.S .S .R .) . The blood coagulation activity of Vipera liberlina turanica venom and its neutralization by specific antiserum Vop. med. Khim. 12, 477, 1966. THE HEMOCOAGULASE activity of the venom from Vipera ßbertina turanica Czernov is an important factor in the pathogenesis of intoxication, particularly immediately following the bite . The problem of suitable qualitative methods for determining coaguaaee activity in snake venoms and anticoagulase activity of sera, did not permit an estimation of the antifermentative actions of antibodies. Also, it has not been possible to determine the anticoagulase activity during purification procedure of sera . The method for determining coaguaaee activity and anticoagulase serum activity, using the statistic method of Kerber, is described. The venom coagulates plasma with sodium citrate by affecting the prothrombin complex; Ca ions are not necessary for this reaction. The optimum pH for coaguaaee activity is between 5-5 and 6-0. Addition of borate anions increased the coagulase activity of the venom. It was possible to separate the coaguaaee and anticoagulase activity using preparative zone electrophoresis. The homologous antiserum antagonized the venom coaguaaee activity. Purification of this antiserum increases the rate of ferment-antiferment action . The anticoagulase and antilethal activity of the serum was in good correlation, and therefore it was possible to use the described method for the determination of antibody level in serum against venom of Yipera libertina. H.R . CHEYMOL, J. (Institut de Pharmacologie, Faculté de Médecine, Paris). De dos substancias biomarinas inhibidoras neuromusculares : tetrodotoxina y saxotoxina . Archos Fac. Med. Madrid 8, 151, 1965 . THE ACTIONS of tetrodotoxin and saxitoxin are compared . In rates the intravenous LDeo are 8 lAgl kg (tetrodo toxin) and 85 jag/kg (saxitoxin). The differences of actions are only quantitative. Tetrodotoxin is more active on nerves than saxitoxin . Saxitoxin is a little more active on neuromuscular preparations, but the effect is more easily reversible . Tetrodotoxin and saxitoxin have an influence on muscle fibres (reducing their excitability), on conduction in nerves, and on the neuromuscular transmission (curarizing action). E .K .
CHEYMOL, J., DEYssoN, G., BOURILLET, F. and ADOLPHE, M. (Institut de Pharmacologie, Faculté de Médecine, Paris). Sur la cytotoxicité de la tetrodotoxine C. r. Séanc. Soc. Biol.159, 1506, 1965 . IN CONTRAST to the high toxicity of tetrodotoxin in mice (intravenous LD bo 11 hg/kg), the toxic effect in tissue culture is very low. Tetrodotoxin acts on metaphase of He La cells in concentrations of only 50 ug/ml. This very low toxicity in tissue culture is not due to thediminuition of toxicity by theculture medium . The experiments prove that tetrodotoxin affects neuromuscular transmission only. The same results were obtained with D-tubocurarine. In mice, the intravenous LDSU for D-tubocurarine is 180 jug/kg; 200 pg/ml were without any effect on He La cells in tissue culture. E.K. DJALDETTI, M., SANDBANK, U., DINsTMAN, M., BIS, H., ROSIN, M. and DE VRIES, A. (Beilinson Hospital, Petah Tikva, Israel). Clot-dissolving action of Echis colorata venom in dogs . Thromb. Math . haemorrh. 16, 420, 1966.
Echis colorata venom, which contains a strong procoaggulant, causes in experimental animals intravascular clotting and subsequent clot dissolution, the latter due to activation of the animal's fibrinolytic system . It was shown that inoculation of Echis colorata venom into dogs is also able to enhance
Abstracts ERspAMER, V., RosEGmNi, M., ENDEAN, R. and ANAsTAsi, A. (Institute of Pharmacology, Univ. of Parma, Parma, Italy ; Dept . of Zoology, Univ . of Queensland, Brisbane, Australia; Research Institute, Farmitalia, Milano, Italy). Biogenic amines and active polypeptides in the skin of Australian amphibians . Natitre Lond. 212, 204, 1966 .
EIGHTEEN species of Australian amphibian were examined . Nine to ten of them were shown to contain considerable amounts of 5-hydroxyindolealkylamines, represented not only by 5-hydroxytryptannine, but also by its N-methyl derivatives. Large amounts of histamine were present only in Hyla caerulea and Hyla infrafrenata . Leptodactyline was always lacking. Physalaemin- and bradykinin-like polypeptides were not detectable, but a new, highly active polypeptide, Caerulein, was discovered in the skin of Hyla caerulea and other species of Hyla. Caerulein possesses, in addition to a powerful, relatively long-lasting hypotensive action, conspicuous effects on a number of external secretions as well as on some extravascular smooth muscles. The possible interest of this research from the view point of biochemical taxonomy is emphasized . V.E .
RtcHARDs, G. M., DU VAIR GENEVIEVE, and LAsxowsxr, SR., M. (Biochemical Laboratory for Cancer Research, Marquette University School of Medicine, Milwaukee, Wisconsin, U.S.A.) . Comparison of the levels of phosphodiesterase, endonuclease, and monophosphatases in several snake venoms . Biochemistry 4, 501, 1965
THE conmoN use of venom phosphodiesterase (PDase) in studying nucleotide sequences requires the use of a preparation of PDase free from contaminating enzymes. In this paper the venoms from five species of snakes were analysed for PDase, 5'-nucleotidase (5'-Nase), endonuclease (ENase), and alkaline nonspecific phosphatase (PMase). The authors were able to determine ENase directly in crude venom by the spectrophotometric method using very dilute solutions of the venom. determination of ENase activity in crude venom had not been successful. Thevenoms of Agkistrodon piscivorus, Bothrops atrox, Crotalus adamanteus, and C. atroxclosely resemble each other in contents and relative proportions of the four enzymes. The PMase activity of the venom of Naja nigricollis is unusually high . Since it is extremely difficult to separate PMase from PDase, Naja venom should be ruled out from the sources for the purification of PDase, a fact recently demonstrated by Russell (Toxicon 4, 153 (1966)). A.O . BRAGANCA, B. M. and KHANDEPARYAR, V. G. (BiochemistryDivision, Indian Cancer Research Centre,Bombay, India) . Phospholipase A activity of cobra venom and lysis of Yoshida sarcoma cells. Life Sciences 5, 1911, 1966 .
A compoNENT of cobra venom labelled P, obtained by column chromatography and fractionation with (NH,),SO, appears to have phospholipase C activity . Incubation of P, with phosphatidyl ethanolamine or phosphotidyl serine, or their lyso derivatives and subsequent analysis by thin-layer chromatography indicated the production of O-phosphoryl ethanolamine and O-phosphoryl serine respectively . The formation of diglycerides could not always be detected perhaps due to traces of phospholipase A and B in P, . Lecithin was resistant to the phospholipase C activity of P, . Phosphatidyl ethanolamine and serine present in the Yoshida sarcoma cells were also susceptible to attack by the P, preparation indicating that observed lysis of these cells by P, may be due to its phospholipase C activity . In mice P, had less than 1 per cent of the toxicity of a purified toxin fraction of cobra venom and less than 0*2 per cent of the hemolytic activity of a purified phospholipase A fraction . P.R . BANNER, A. H., HoLFmcH, P. and PrYAKARNCHANA, T. (Hawaii Marine Laboratory, University of Hawaii, Honolulu, Hawaii). Retention of ciguatera toxin by the red snapper, Lutjanus bohar. Copela 2, 297, 1966.
A sAMPLE of 66 samples of the red snapper, Lutjanus bohar, from Christmas Island, Line Islands, whose initial level of toxicity was approximated statistically, was maintained in the holding ponds at the Hawaii Marine Laboratory, Honolulu, for varying periods from 3 to 30 months on a nontoxic diet . At the end of the period the fish showed no significant decline in their toxicity . (Authors' abstract).
70
Abstracts
This work establishes the feasibility of the food chain hypothesis in ciguatera intoxication after the ingestion of fish because it shows effectively that the toxin level in the tissue of fish maintained for long periods of time on a nontoxic diet does not diminish . J.F .G . RorrMAN, M. I. (Science Research Institute Vaccines, Minister Public Health, Taschkent, U.Z.S .S .R .) . The blood coagulation activity of Vipera liberlina turanica venom and its neutralization by specific antiserum Vop. med. Khim. 12, 477, 1966. THE HEMOCOAGULASE activity of the venom from Vipera ßbertina turanica Czernov is an important factor in the pathogenesis of intoxication, particularly immediately following the bite . The problem of suitable qualitative methods for determining coaguaaee activity in snake venoms and anticoagulase activity of sera, did not permit an estimation of the antifermentative actions of antibodies. Also, it has not been possible to determine the anticoagulase activity during purification procedure of sera . The method for determining coaguaaee activity and anticoagulase serum activity, using the statistic method of Kerber, is described. The venom coagulates plasma with sodium citrate by affecting the prothrombin complex; Ca ions are not necessary for this reaction. The optimum pH for coaguaaee activity is between 5-5 and 6-0. Addition of borate anions increased the coagulase activity of the venom. It was possible to separate the coaguaaee and anticoagulase activity using preparative zone electrophoresis. The homologous antiserum antagonized the venom coaguaaee activity. Purification of this antiserum increases the rate of ferment-antiferment action . The anticoagulase and antilethal activity of the serum was in good correlation, and therefore it was possible to use the described method for the determination of antibody level in serum against venom of Yipera libertina. H.R . CHEYMOL, J. (Institut de Pharmacologie, Faculté de Médecine, Paris). De dos substancias biomarinas inhibidoras neuromusculares : tetrodotoxina y saxotoxina . Archos Fac. Med. Madrid 8, 151, 1965 . THE ACTIONS of tetrodotoxin and saxitoxin are compared . In rates the intravenous LDeo are 8 lAgl kg (tetrodo toxin) and 85 jag/kg (saxitoxin). The differences of actions are only quantitative. Tetrodotoxin is more active on nerves than saxitoxin . Saxitoxin is a little more active on neuromuscular preparations, but the effect is more easily reversible . Tetrodotoxin and saxitoxin have an influence on muscle fibres (reducing their excitability), on conduction in nerves, and on the neuromuscular transmission (curarizing action). E .K .
CHEYMOL, J., DEYssoN, G., BOURILLET, F. and ADOLPHE, M. (Institut de Pharmacologie, Faculté de Médecine, Paris). Sur la cytotoxicité de la tetrodotoxine C. r. Séanc. Soc. Biol.159, 1506, 1965 . IN CONTRAST to the high toxicity of tetrodotoxin in mice (intravenous LD bo 11 hg/kg), the toxic effect in tissue culture is very low. Tetrodotoxin acts on metaphase of He La cells in concentrations of only 50 ug/ml. This very low toxicity in tissue culture is not due to thediminuition of toxicity by theculture medium . The experiments prove that tetrodotoxin affects neuromuscular transmission only. The same results were obtained with D-tubocurarine. In mice, the intravenous LDSU for D-tubocurarine is 180 jug/kg; 200 pg/ml were without any effect on He La cells in tissue culture. E.K. DJALDETTI, M., SANDBANK, U., DINsTMAN, M., BIS, H., ROSIN, M. and DE VRIES, A. (Beilinson Hospital, Petah Tikva, Israel). Clot-dissolving action of Echis colorata venom in dogs . Thromb. Math . haemorrh. 16, 420, 1966.
Echis colorata venom, which contains a strong procoaggulant, causes in experimental animals intravascular clotting and subsequent clot dissolution, the latter due to activation of the animal's fibrinolytic system . It was shown that inoculation of Echis colorata venom into dogs is also able to enhance