Toxic, antimicrobial and hemagglutinating activities of the purple fluid of the sea hare Aplysia dactylomela Rang, 1828

Melo, Vânia M. M.; Fonseca, Andre; Vasconcelos, Ilka M.; Carvalho, A. C. P. P. de

doi:10.1590/s0100-879x1998000600010

Cited by 15 publications

(9 citation statements)

References 28 publications

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“…Unlike previous reports of similar proteins isolated from sea hares (Kisugi et al, 1989;Yamazaki et al, 1990;Melo et al, 1998Melo et al, , 2000, we demonstrate that substrates determined the bactericidal or bacteriostatic effects of escapin. Bacteria cultured in M9 and glucose medium without high levels (50·mmol·l -1 ) of selected amino acids or without Tryptone Peptone supplements were not killed (Figs·7, 8).…”

Section: Discussioncontrasting

confidence: 99%

“…Several of these proteins, including three from sea hares, are known to be L-amino acid oxidases (Iijima et al, 2003;Jimbo et al, 2003;Butzke et al, 2004), but the cofactors responsible for oxidase activity have not been experimentally demonstrated. These studies differ in their findings as to whether the proteins are bacteriostatic or bactericidal (Yamazaki et al, 1990;Otsuka-Fuchino et al, 1992;Melo et al, 1998;. In addition to the interesting biological roles these proteins may play for their host organisms, some may also have practical industrial or medical uses, yet none have been successfully expressed in high output systems such as bacteria (Obara et al, 1999;Torii et al, 2000;Petzelt et al, 2002;Butzke et al, 2004;Cummins et al, 2004).…”

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Cloning, characterization and expression of escapin, a broadly antimicrobial FAD-containing l-amino acid oxidase from ink of the sea hare Aplysia californica

Yang

Johnson

et al. 2005

Journal of Experimental Biology

107

122

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SUMMARY A 60 kDa monomeric protein isolated from the defensive purple ink secretion of the sea hare Aplysia californica was cloned and sequenced, and is the first sea hare antimicrobial protein to be functionally expressed in E. coli. Sequence analysis suggested that this protein is a flavin-containing l-amino acid oxidase (LAAO), with one predicted potential glycosylation site, although the glycosylation could not be experimentally confirmed. This protein, which we call `escapin', has high sequence similarity to several other gastropod proteins. Escapin was verified by NMR, mass spectroscopy and HPLC to have FAD as its flavin cofactor. Escapin's antimicrobial effects, bacteriostasis and bactericidal, were determined using a combination of two assays: (1) incubation of bacteria on solid media followed by assessment of inhibition by direct observation of zones of inhibition or by turbidity measurements; and (2) incubation of bacteria in liquid media followed by counting viable colonies after growing on agar plates. Native escapin inhibited the growth of Gram-positive and Gram-negative bacteria, including marine bacteria (Vibrio harveyiiand Staphylococcus aureus) and pathogenic bacteria(Staphylococcus aureus, Streptococcus pyogenes and Pseudomonas aeruginosa). Escapin also inhibited the growth of yeast and fungi, with different efficacies. Escapin's antimicrobial activity was concentration dependent and did not decrease when stored for more than 5 months at room temperature. Escapin was bacteriostatic and not bactericidal in minimal media (e.g. salt media) with glucose, yeast extract, and a mixture of 20 amino acids each at 50 μmol l-1, but was bactericidal in media enriched with Tryptone Peptone. Escapin was also strongly bactericidal in media with l-lysine at concentrations as low as 3 mmol l-1 and slightly bactericidal in 50 mmol l-1l-arginine, but not in most other amino acids even at 50 mmol l-1. Escapin had high oxidase activity (producing hydrogen peroxide) with either l-arginine or l-lysine as a substrate and little to no oxidase activity with other l-amino acids. Hydrogen peroxide alone (without escapin or amino acids) was strongly bacteriostatic but poorly bactericidal, similar in this respect to l-arginine but different from l-lysine in the presence of escapin. Together these results suggest that there are multiple mechanisms to escapin's antimicrobial effects, with bacteriostasis resulting largely or entirely from the effects of hydrogen peroxide produced by escapin's LAAO activity, but bactericidal effects resulting from lysine-dependent mechanisms not directly involving hydrogen peroxide. Recombinant escapin expressed in bacteria was also active against Gram-positive and Gram-negative bacteria,suggesting that glycosylation is not essential for antimicrobial activity.

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Cloning, characterization and expression of escapin, a broadly antimicrobial FAD-containing l-amino acid oxidase from ink of the sea hare Aplysia californica

Yang

Johnson

et al. 2005

Journal of Experimental Biology

107

122

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“…The neuroexcitatory signs are consistent with those seen in other domestic species that have had toxicosis induced by chemical extracts from related sea hares. 5,6,7,8 Toxicosis may be due to the presence of cholinomimetic chemicals in the digestive gland and such chemicals have been isolated from other Aplysia species. 9,10,11,12,13 Gastrointestinal and respiratory abnormalities were also identified in the present study.…”

Section: Discussionmentioning

confidence: 99%

“…Several studies have reported respiratory distress prior to death in mice injected intraperitoneally with extracts from other Aplysia species. 7,8 The cause of respiratory distress is unknown, however, bronchoconstriction may play a role if the toxin has cholinomimetic effects. 9,10,11,12,13 In select cases, the use of a bronchodilator may be justified.…”

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confidence: 99%

“…Studies investigating the toxic effects of chemical extracts from other sea hare species only report clinical signs of the toxicosis and the cause of death is unknown. 5,6,7,8 The reason stated for euthanasia, where performed, was a worsening of clinical signs despite treatment. This study may misrepresent the number of dogs that died, or the proportion of dogs that developed clinical signs, after exposure to A. gigantea as there may be a population of dogs with exposure that never presented to a veterinarian.…”

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Aplysia gigantea toxicosis in 72 dogs in Western Australia

et al. 2013

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Objectives This study aimed to: (1) confirm a temporal association between exposure to the sea hare Aplysia gigantea and the development of a neurotoxicosis in dogs and (2) further characterise the clinical signs, treatment and outcomes in dogs with this suspected toxicosis. Methods The medical records from four veterinary practices within the Geraldton region of Western Australia were searched for dogs that had been exposed to A. gigantea and subsequently presented to a veterinarian during the period of January 2001 to March 2011. Signalment, exposure history, clinical signs, treatment and outcome were recorded. Results In total, 72 dogs met the inclusion criteria. Clinical signs included ptyalism, emesis, ataxia, hyperaesthesia, tremors, muscle fasciculations, seizures, nystagmus and respiratory distress; 30 dogs did not have abnormal clinical signs at presentation; 69 dogs were presented during January to April. Treatment included gastrointestinal and dermal decontamination, and supportive management of seizures, tremors and muscle fasciculations. Of the 72 dogs, 65 survived to discharge, 4 died and 3 were euthanased. Information from subsequent examinations was available for 57 dogs and no long‐term complications were reported. Conclusions Exposure to A. gigantea was temporally associated with the development of neuroexcitatory clinical signs in dogs. Gastrointestinal and respiratory signs also occurred in some dogs. Dogs with suspected toxicosis were presented mostly in the months of January to April. The proportion of dogs that died or were euthanased because of worsening clinical signs was approximately 10%.

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Mollusks–1

Kornprobst

2014

Encyclopedia of Marine Natural Products

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Toxic, antimicrobial and hemagglutinating activities of the purple fluid of the sea hare Aplysia dactylomela Rang, 1828

Cited by 15 publications

References 28 publications

Cloning, characterization and expression of escapin, a broadly antimicrobial FAD-containing l-amino acid oxidase from ink of the sea hare Aplysia californica

Cloning, characterization and expression of escapin, a broadly antimicrobial FAD-containing l-amino acid oxidase from ink of the sea hare Aplysia californica

Aplysia gigantea toxicosis in 72 dogs in Western Australia

Mollusks–1

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