Poisoning after ingestion of pufferfish in Brazil: report of 11 cases

In this brief communication the authors report eleven cases of human poisoning caused by ingestion of pufferfish meat. Three patients (two children and one adult) were seriously affected. The circumstances that precipitated the poisoning are discussed as well as the clinical aspects observed. No deaths were registered and the patients did not present sequelae after the episode.(AU)

Poisoning after ingestion of pufferfish in Brazil: report of 11 cases

In this brief communication the authors report eleven cases of human poisoning caused by ingestion of pufferfish meat. Three patients (two children and one adult) were seriously affected. The circumstances that precipitated the poisoning are discussed as well as the clinical aspects observed. No deaths were registered and the patients did not present sequelae after the episode.

Systemic effects induced by intralesional injection of ?-conotoxin MVIIC after spinal cord injury in rats

Calcium channel blockers such as conotoxins have shown a great potential to reduce brain and spinal cord injury. MVIIC neuroprotective effects analyzed in in vitro models of brain and spinal cord ischemia suggest a potential role of this toxin in preventing injury after spinal cord trauma. However, previous clinical studies with MVIIC demonstrated that clinical side effects might limit the usefulness of this drug and there is no research on its systemic effects. Therefore, the present study aimed to investigate the potential toxic effects of MVIIC on organs and to evaluate clinical and blood profiles of rats submitted to spinal cord injury and treated with this marine toxin. Rats were treated with placebo or MVIIC (at doses of 15, 30, 60 or 120 pmol) intralesionally following spinal cord injury. Seven days after the toxin administration, kidney, brain, lung, heart, liver, adrenal, muscles, pancreas, spleen, stomach, and intestine were histopathologically investigated. In addition, blood samples collected from the rats were tested for any hematologic or biochemical changes.(AU)

Systemic effects induced by intralesional injection of ?-conotoxin MVIIC after spinal cord injury in rats

Calcium channel blockers such as conotoxins have shown a great potential to reduce brain and spinal cord injury. MVIIC neuroprotective effects analyzed in in vitro models of brain and spinal cord ischemia suggest a potential role of this toxin in preventing injury after spinal cord trauma. However, previous clinical studies with MVIIC demonstrated that clinical side effects might limit the usefulness of this drug and there is no research on its systemic effects. Therefore, the present study aimed to investigate the potential toxic effects of MVIIC on organs and to evaluate clinical and blood profiles of rats submitted to spinal cord injury and treated with this marine toxin. Rats were treated with placebo or MVIIC (at doses of 15, 30, 60 or 120 pmol) intralesionally following spinal cord injury. Seven days after the toxin administration, kidney, brain, lung, heart, liver, adrenal, muscles, pancreas, spleen, stomach, and intestine were histopathologically investigated. In addition, blood samples collected from the rats were tested for any hematologic or biochemical changes.

Ômega - conotoxina MVIIC no tratamento do trauma medular agudo em ratos wistar.

O influxo de cálcio é um mecanismo importante de lesão secundária do trauma medular agudo (TMA). Pesquisas com bloqueadores de canais para cálcio vêm sendo desenvolvidas com intuito de estabelecerem novas terapias para reduzir lesões secundárias ao trauma medular. Toxinas oriundas de veneno de caramujo marinho se mostram como terapia promissora por inibirem seletivamente os canais para cálcio e impedirem o influxo desse íon, o que reduziria a progressão da lesão. Dentre essas toxinas, a -conotoxina MVIIC, do caramujo Conus magus, tem sido utilizada por inibir seletivamente canais para cálcio, impedindo a progressão da lesão e atuando de forma benéfica no tratamento do TMA. O objetivo desse estudo foi avaliar o efeito da -conotoxina MVIIC na medula de ratos submetidos a TMA por meio da análise de fatores de neuroproteção como extresse oxidativo e apoptose celular, efeitos sistêmicos da -conotoxina MVIIC, além da função locomotora. Foram utilizados 36 ratos Wistar, machos, adultos, pesando em média 450g, distribuídos aleatoriamente em seis grupos de seis animais: controle positivo (CTL+), controle negativo (CTL -), TOX 20 pMOL 30, TOX 20 pMOL 2h, TOX 40 pMOL 30, TOX 40 pMOL 2h. Todos os animais foram submetidos a laminectomia da vértebra T12 e TMA, sendo que o grupo CTL - não foi submetido ao trauma após laminectomia. As lesões foram produzidas obedecendo ao protocolo internacional MASCIS (Multicenter Animal Spinal Cord Injury Study), com a liberação em queda livre de haste de impacto de peso de 10 g, com altura pré-determinada de 12.5 mm sobre a dura-máter por três segundos. Após 30 minutos e duas horas do trauma inicial, o CTL+ recebeu injeção intratecal de 10µL de PBS estéril e os grupos TOX, volume equivalente de solução de toxina MVIIC na concentração preconizada para cada grupo. Foram observadas recuperação locomotora, redução da expressão gênica de fatores proapoptóticos e aumento da expressão gênica de fator anti apoptótico, além de redução de estresse oxidativo em ratos tratados com MVIIC pós TMA, o que sugere efeito neuroprotetor dessa toxina na medula espinhal

Calcium influx is an important mechanism of secondary injury to acute medullary trauma (TMA). Studies with calcium channel blockers have been developed in order to establish new therapies to reduce lesions secondary to spinal cord trauma. Toxins derived from marine snail venom are shown as promising therapy by selectively inhibiting the calcium channels and preventing the influx of this ion, which would reduce the progression of the lesion. Thus, -conotoxin MVIIC has been used by selectively inhibiting calcium channels, preventing the progression of the lesion and may have a beneficial effect for the treatment of acute medullary trauma . The objective of this study was to evaluate neuroprotection factors such as lipid peroxidation, oxidative stress and cellular apoptosis, systemic effects of -conotoxin MVIIC, as well as the locomotor function of rats submitted to TMA and treated with MVIIC conotoxin. METHODS: 36 male Wistar rats weighing 450g were randomly assigned to six groups of six animals: positive control (CTL +), negative control (CTL -), TOX 20 pMOL 30 ', TOX 20 pMOL 2h, TOX 40 pMOL 30 ', TOX 40 pMOL 2h. All the animals underwent laminectomy of the T12 vertebra and TMA, except for the CTL group - who did not suffer the trauma. The lesions were produced according to the international protocol MASCIS (Multicenter Animal Spinal Cord Injury Study), with the release in free drop of 10 g weight impact, with a predetermined height of 12.5 mm on the dura mater for 3 seconds . After 30 minutes and 2 hours of initial trauma, CTL + received intrathecal injection of 10L of sterile PBS and TOX groups, equivalent volume of MVIIC toxin solution at the recommended concentration for each group. It was observed that locomotor recovery and neuroprotection mediated by reduction of the gene expression of pro-apoptotic factors and increased gene expression of anti-apoptotic factor by means of the qRT-PCR technique in addition to reduction of oxidative stress in rats treated with MVIIC after acute medullary injury.

Biomedical and pharmacological potential of tetrodotoxin-producing bacteria isolated from marine pufferfish Arothron hispidus (Muller, 1841)

Specimens of the pufferfish Arothron hispidus collected at Parangipettai, on the southeast coast of India, were subjected to bacterial isolation and identification. Three species were identified, namely Bacillus sp., Kytococcus sedentarius and Cellulomonas fimi. Partially-purified microbial filtrates exhibited hemolytic activity on chicken and human erythrocytes of O, B and AB blood groups, with maximum activity of 32 HU. The microbial filtrates also presented ATPase, Mg2+-ATPase, Na+K+-ATPase and AchE enzymatic activities of positive neuromodulation in Kytococcus sedentarius with 1300, 300.1, 1549.98 and 140.55%, in Cellulomonas fimi with 620, 300, 10 and 128.42%, and in Bacillus species with 40, 200, 849.98 and 158.69%, respectively. Toxicity symptoms were observed when the bacterial filtrate was intraperitoneally injected into mice. The bacterial filtrate caused adverse effects on viability of the mouse muscle cell line (L929) and leukemia cell line (P388). Maximum level of inhibition was observed on the growth of L929 cell line. Bacillus lentimorbus inhibited the cell line from 84.03 to 94.43% whereas Bacillus species inhibited the growth in a range between 77.25 and 86.16% at the lowest dilution.Specimens of the pufferfish Arothron hispidus collected at Parangipettai, on the southeast coast of India, were subjected to bacterial isolation and identification. Three species were identified, namely Bacillus sp., Kytococcus sedentarius and Cellulomonas fimi. Partially-purified microbial filtrates exhibited hemolytic activity on chicken and human erythrocytes of O, B and AB blood groups, with maximum activity of 32 HU. The microbial filtrates also presented ATPase, Mg2+-ATPase, Na+K+-ATPase and AchE enzymatic activities of positive neuromodulation in Kytococcus sedentarius with 1300, 300.1, 1549.98 and 140.55 percent, in Cellulomonas fimi with 620, 300, 10 and 128.42 percent, and in Bacillus species with 40, 200, 849.98 and 158.69 percent, respectively. Toxicity symptoms were observed when the bacterial filtrate was intraperitoneally injected into mice. The bacterial filtrate caused adverse effects on viability of the mouse muscle cell line (L929) and leukemia cell line (P388). Maximum level of inhibition was observed on the growth of L929 cell line. Bacillus lentimorbus inhibited the cell line from 84.03 to 94.43 percent whereas Bacillus species inhibited the growth in a range between 77.25 and 86.16 percent at the lowest dilution.(AU)

Biomedical and pharmacological potential of tetrodotoxin-producing bacteria isolated from marine pufferfish Arothron hispidus (Muller, 1841)

Specimens of the pufferfish Arothron hispidus collected at Parangipettai, on the southeast coast of India, were subjected to bacterial isolation and identification. Three species were identified, namely Bacillus sp., Kytococcus sedentarius and Cellulomonas fimi. Partially-purified microbial filtrates exhibited hemolytic activity on chicken and human erythrocytes of O, B and AB blood groups, with maximum activity of 32 HU. The microbial filtrates also presented ATPase, Mg2+-ATPase, Na+K+-ATPase and AchE enzymatic activities of positive neuromodulation in Kytococcus sedentarius with 1300, 300.1, 1549.98 and 140.55%, in Cellulomonas fimi with 620, 300, 10 and 128.42%, and in Bacillus species with 40, 200, 849.98 and 158.69%, respectively. Toxicity symptoms were observed when the bacterial filtrate was intraperitoneally injected into mice. The bacterial filtrate caused adverse effects on viability of the mouse muscle cell line (L929) and leukemia cell line (P388). Maximum level of inhibition was observed on the growth of L929 cell line. Bacillus lentimorbus inhibited the cell line from 84.03 to 94.43% whereas Bacillus species inhibited the growth in a range between 77.25 and 86.16% at the lowest dilution.(AU)

Cianobacterial bloom and animal mass mortality in a reservoir from Central Argentina / Florescimento de cianobactérias e mortalidade em massa de animais em um reservatório da Argentina Central

Piedras Moras reservoir (32° 10'27" S and 64° 16' 29" W; 832 ha), integrates a series of artificial lakes belonging to the Rio Tercero basin (Córdoba, Argentina). During March 2009 an algal bloom occurred, coinciding with several animal species mortality, mainly wild birds. The goal of this work was to establish the trophic status of the reservoir in relation to that mortality. Variables were evaluated in situ (temperature and water transparency) and samples were taken in order to identify algal species, Chl-a concentration (spectrophotometry) and toxins - total microcystines- (inmuno-enzymatic assay, ELISA). Histopathology studies were made on Fulica sp. A strong heterogenity in water transparency was observed, and "patches" of Potamogeton berteroanus distributed all along the lake, with Secchi disk minimal and medium values of 0.15 and 0.94 m. Chl-a concentration oscillated from 35.7 to 320.9 mg.m-3. Predominant phytoplankton species were Anabaena spiroides and Microcystis aeruginosa (Cyanophyceae). Water temperature was 27.8 °C (±0.88). Maximal value of total microcystine concentration was 0.23 μg.L-1. Chl-a concentration at the moment when mass mortality occurred (2.022 mg.m-3), and histopathological observations, strongly suggest that the animals' death was due to cianotoxins.

O reservatório de Piedras Moras (32° 10'27" S e 64° 16' 29" W; 832 ha) faz parte de uma série de lagos artificiais que estão localizados na bacia do rio Tercero (Córdoba, Argentina). Durante março de 2009, ocorreu um florescimento maciço de cianobactérias que resultou na morte de muitas espécies de animais, especialmente aves aquáticas silvestres. O objetivo deste trabalho foi o de estabelecer o estado trófico do reservatório em função desta mortalidade. As variáveis foram avaliadas in situ (temperatura e transparência da água). Foram coletadas amostras para analisar as espécies de algas encontradas nos florescimentos. Foram determinados chl-a (espectrofotometria) e toxinas (microcistinas totais - ensaio imunoenzimático ELISA). Estudos histopalógicos foram realizados em Fulica sp. Uma grande heterogeneidade em transparência de água foi observada e agrupamentos de Potamogeton berteroanus estavam distribuídos em todo o reservatório com valores mínimos e médios do disco de Secchi de 0,15 a 0,54 m. Concentrações de chl-a oscilaram de 35,7 a 320,9 mg.m-3. As espécies fitoplanctônicas dominantes identificadas foram Anabaena spiroides e Microcystis aeruginosa (Cyanophyceae). A temperatura da água foi de 27,8 °C (±0.88). O valor máximo de concentração de microcistinas foi de 0,23 µg.L-1. A concentração de chl-a, quando ocorreu a mortalidade em massa (2,022 mg.m-3), e as lesões histopatológicas indicam que a mortalidade de animais foi devida a cianotoxinas.

Characterization, purification and phylogenetic analysis of a cytolysin from the sea anemone Heteractis magnifica of the Indian Ocean

It is well established that sea anemones comprise a rich source of cytolytic toxins. The present study reports the isolation and characterization of a cytolysin obtained from the sea anemone Heteractis magnifica collected in the Andaman Islands of the Indian Ocean. The crude extract was screened for hemolytic activity by a blood agar plate method and a 6-mm zone of clearance was observed after incubation. The hemolytic property of the crude extract, tested by the microtiter plate method, revealed positive results at concentrations as low as 120 ng/mL. Furthermore, it was favored by alkaline pH and was stable up to 60°C. On the other hand, the hemolytic effect was abolished by the addition of human serum. Purification steps involved ammonium sulfate precipitation and subsequent desalting by dialysis, followed by anion- and cation-exchange chromatographies. The purified fractions displayed the presence of a 19-kDa cytolysin when analyzed by SDS-PAGE. The conserved region of the cytolysin (with 303 bp) was amplified by RT-PCR and was sequenced. The sequence showed maximum homology (97 percent) with the already reported cytolysins from other sea anemone species.(AU)

Characterization, purification and phylogenetic analysis of a cytolysin from the sea anemone Heteractis magnifica of the Indian Ocean

It is well established that sea anemones comprise a rich source of cytolytic toxins. The present study reports the isolation and characterization of a cytolysin obtained from the sea anemone Heteractis magnifica collected in the Andaman Islands of the Indian Ocean. The crude extract was screened for hemolytic activity by a blood agar plate method and a 6-mm zone of clearance was observed after incubation. The hemolytic property of the crude extract, tested by the microtiter plate method, revealed positive results at concentrations as low as 120 ng/mL. Furthermore, it was favored by alkaline pH and was stable up to 60ºC. On the other hand, the hemolytic effect was abolished by the addition of human serum. Purification steps involved ammonium sulfate precipitation and subsequent desalting by dialysis, followed by anion- and cation-exchange chromatographies. The purified fractions displayed the presence of a 19-kDa cytolysin when analyzed by SDS-PAGE. The conserved region of the cytolysin (with 303 bp) was amplified by RT-PCR and was sequenced. The sequence showed maximum homology (97 percent) with the already reported cytolysins from other sea anemone species.(AU)

Bioactivities of extracts from the marine sponge Halichondria panicea

In the present study, we screened the biological activity of extracts from the marine sponge Halichondria panicea collected in the Arabian Sea. Crude toxin was obtained by methanol, chloroform-methanol (2:1) and aqueous extraction. Subsequently, the protein concentration of each crude extract was determined. The impact of both sponge methanolic and aqueous extracts was found to increase activities of Na+-K+ ATP-ase and Mg++ ATP-ase. In the case of chloroform-methanol extract, higher concentrations increased acetylcholine esterase (AchE) activity. The methanolic and chloroform-methanol extracts exhibited hemolytic activity on chicken and human erythrocytes, whereas the aqueous extract failed to do so. Methanol and aqueous extracts produced an immunostimulating effect and all extracts revealed angiogenic activity. The aqueous extract yielded nine bands by SDS-PAGE on 12% gel.

Bioactivities of extracts from the marine sponge Halichondria panicea

In the present study, we screened the biological activity of extracts from the marine sponge Halichondria panicea collected in the Arabian Sea. Crude toxin was obtained by methanol, chloroform-methanol (2:1) and aqueous extraction. Subsequently, the protein concentration of each crude extract was determined. The impact of both sponge methanolic and aqueous extracts was found to increase activities of Na+-K+ ATP-ase and Mg++ ATP-ase. In the case of chloroform-methanol extract, higher concentrations increased acetylcholine esterase (AchE) activity. The methanolic and chloroform-methanol extracts exhibited hemolytic activity on chicken and human erythrocytes, whereas the aqueous extract failed to do so. Methanol and aqueous extracts produced an immunostimulating effect and all extracts revealed angiogenic activity. The aqueous extract yielded nine bands by SDS-PAGE on 12% gel.(AU)

Botulismo em alimentos: um problema de saúde pública / Botulism in sustenances: a problem of public health

O Clostridium botulinum é uma bactéria esporogênica, anaeróbia, comumente encontrada em solos, sedimentos marinhos e de águas doces, bem como no trato intestinal de homens e animais. Condições industriais inadequadas podem ocasionar o seu desenvolvimento e este, por sua vez, produz toxina se lhe forem dadas condições como potencial redox, pH, temperatura e umidade. As maiores fontes de contaminação são os legumes (57 por cento) , pescados (15 por cento), frutas em conserva e condimentos, incluindo o mel (8 por cento). Suas toxinas são classificadas, de acordo com a especificidade sorológica, em sete tipos, caracterizados pelas letras de A a G. Também de acordo com a homologia dos valores de DNA e RNA de cada grupo, bem como na uniformidade dos produtos pirolíticos, podem ser subdivididas em quatro grupos distintos. Diversas cepas produzem neurotoxinas; entretanto, os efeitos farmacológicos são similares mesmo possuindo estruturas protéicas diferentes. Todas as cepas são mesófilas, com crescimento ideal entre 25ºC e 37ºC, e se desenvolvem melhor em pH próximo à neutralidade, tendendo ao alcalino (7,0 a 7,4). Entretanto, a do tipo E pode se desenvolver e produzir toxina em temperaturas muito baixas, assim como certas cepas são passíveis de se desenvolver em pH mais baixo. A contaminação dos animais domésticos ocorre freqüentemente a partir de cadáveres de roedores, animais silvestres, aves, silagens, pastos ou bebedouros contaminados. Para o homem, alimentos de origem animal como carnes e seus derivados, estão entre os mais incriminados na veiculação de C. botulinum, sobretudo aqueles produtos artesanais, ou mesmo os inspecionados onde houve falhas em uma ou mais etapas do processamento tecnológico.(AU)

Clostridium botulinum is an anaerobic sporougenous germ usually found in soil, marine and fresh water sediments, as well as in the intestinal tract of humans and animals. In industrialized food products, favorable conditions of pH, temperature and oxidation-reduction potential can facilitate its development and growing. The main contaminations sources are vegetable (57%),fish (15%), canned fruits, and seasoning, including honey (8%). The Clostridium are classified according with its serologic specificity, in seven different serotypes from A to G). Besides, in accordance with DNA and RNA homology and uniformity of pirolitics products, it can be subdivide in four distinct groups. Different strains produce different neurotoxins, but the pharmacological effects are similar, even if the proteic chain structure is unlike. All the strains are mesophillics, with the ideal temperature of growing from 25°C to 37°C and the neutral pH tending to alkaline (7,0 - 7,4). Nevertheless, the E strain can develop produce toxins in very low temperatures and some can develop in lower pH. The domestic animals contamination occurs usually from feeding rodent, avian and wild animals dead bodies, contaminated pasture, drinking water and silage. For the man, the main contamination source is food, like meat, seafood and its products, mainly the handcraft products or when the inspection fails in one or more stages of technologic processment. (AU)

Botulismo em alimentos: um problema de saúde pública / Botulism in sustenances: a problem of public health

O Clostridium botulinum é uma bactéria esporogênica, anaeróbia, comumente encontrada em solos, sedimentos marinhos e de águas doces, bem como no trato intestinal de homens e animais. Condições industriais inadequadas podem ocasionar o seu desenvolvimento e este, por sua vez, produz toxina se lhe forem dadas condições como potencial redox, pH, temperatura e umidade. As maiores fontes de contaminação são os legumes (57 por cento) , pescados (15 por cento), frutas em conserva e condimentos, incluindo o mel (8 por cento). Suas toxinas são classificadas, de acordo com a especificidade sorológica, em sete tipos, caracterizados pelas letras de A a G. Também de acordo com a homologia dos valores de DNA e RNA de cada grupo, bem como na uniformidade dos produtos pirolíticos, podem ser subdivididas em quatro grupos distintos. Diversas cepas produzem neurotoxinas; entretanto, os efeitos farmacológicos são similares mesmo possuindo estruturas protéicas diferentes. Todas as cepas são mesófilas, com crescimento ideal entre 25ºC e 37ºC, e se desenvolvem melhor em pH próximo à neutralidade, tendendo ao alcalino (7,0 a 7,4). Entretanto, a do tipo E pode se desenvolver e produzir toxina em temperaturas muito baixas, assim como certas cepas são passíveis de se desenvolver em pH mais baixo. A contaminação dos animais domésticos ocorre freqüentemente a partir de cadáveres de roedores, animais silvestres, aves, silagens, pastos ou bebedouros contaminados. Para o homem, alimentos de origem animal como carnes e seus derivados, estão entre os mais incriminados na veiculação de C. botulinum, sobretudo aqueles produtos artesanais, ou mesmo os inspecionados onde houve falhas em uma ou mais etapas do processamento tecnológico.

Clostridium botulinum is an anaerobic sporougenous germ usually found in soil, marine and fresh water sediments, as well as in the intestinal tract of humans and animals. In industrialized food products, favorable conditions of pH, temperature and oxidation-reduction potential can facilitate its development and growing. The main contaminations sources are vegetable (57%),fish (15%), canned fruits, and seasoning, including honey (8%). The Clostridium are classified according with its serologic specificity, in seven different serotypes from A to G). Besides, in accordance with DNA and RNA homology and uniformity of pirolitics products, it can be subdivide in four distinct groups. Different strains produce different neurotoxins, but the pharmacological effects are similar, even if the proteic chain structure is unlike. All the strains are mesophillics, with the ideal temperature of growing from 25°C to 37°C and the neutral pH tending to alkaline (7,0 - 7,4). Nevertheless, the E strain can develop produce toxins in very low temperatures and some can develop in lower pH. The domestic animals contamination occurs usually from feeding rodent, avian and wild animals dead bodies, contaminated pasture, drinking water and silage. For the man, the main contamination source is food, like meat, seafood and its products, mainly the handcraft products or when the inspection fails in one or more stages of technologic processment.