A protein phosphatase 2A inhibition assay for a fast and sensitive assessment of okadaic acid contamination in mussels

Tubaro, Aurelia; Florio, Chiara; Luxich, Elena; Sosa, Silvio; Loggia, R. Della; Yasumoto, Takeshi

doi:10.1016/0041-0101(96)00027-x

Cited by 138 publications

(67 citation statements)

References 14 publications

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“…Tissue samples were also tested for brevetoxin (North Carolina State University, Raleigh) using micellar electrokinetic capillary chromatography with laser-induced fluorescence detection (MEKC-LIF; Shea 1997). Extracts were also analyzed for okadaic acid (and other protein phosphatase inhibitors) using a colorimetric protein-phosphatase inhibition assay modified from Tubaro et al (1996). Portions of stomach contents and urine samples were screened for domoic acid and palytoxin (NOAA).…”

Section: Methodsmentioning

confidence: 99%

Neurological disease in wild loggerhead sea turtles Caretta caretta

Jacobson

Homer²,

Stacy³

et al. 2006

Dis. Aquat. Org.

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Beginning in October 2000, subadult loggerhead sea turtles Caretta caretta showing clinical signs of a neurological disorder were found in waters off south Florida, USA. Histopathology indicated generalized and neurologic spirorchiidiasis. In loggerhead sea turtles (LST) with neurospirorchiidiasis, adult trematodes were found in the meninges of the brain and spinal cord of 7 and 3 affected turtles respectively, and multiple encephalic intravascular or perivascular eggs were associated with granulomatous or mixed leukocytic inflammation, vasculitis, edema, axonal degeneration and occasional necrosis. Adult spirorchiids were dissected from meningeal vessels of 2 of 11 LST brains and 1 of 10 spinal cords and were identified as Neospirorchis sp. Affected LST were evaluated for brevetoxins, ciguatoxins, saxitoxins, domoic acid and palytoxin. While tissues from 7 of 20 LST tested positive for brevetoxins, the levels were not considered to be in a range causing acute toxicosis. No known natural (algal blooms) or anthropogenic (pollutant spills) stressors co-occurred with the turtle mortality. While heavy metal toxicosis and organophosphate toxicosis were also investigated as possible causes, there was no evidence for their involvement. We speculate that the clinical signs and pathologic changes seen in the affected LST resulted from combined heavy spirorchiid parasitism and possible chronic exposure to a novel toxin present in the diet of LST.KEY WORDS: Spirorchiidiasis · Brain · Spinal cord · Neuropathy · Loggerhead sea turtle · Caretta caretta Resale or republication not permitted without written consent of the publisherDis Aquat Org 70: [139][140][141][142][143][144][145][146][147][148][149][150][151][152][153][154] 2006 be under-diagnosed rather than being a rare occurrence.Digenetic trematodes of the family Spirorchiidae utilize freshwater turtles and sea turtles as their definitive hosts, and are known to cause neurologic complications. At least 8 genera and 20 species of spirorchiids infect the loggerhead Caretta caretta, green Chelonia mydas and hawksbill Eretmochelys imbricata sea turtles (Lauckner 1985), and are the most pathogenic of sea turtle parasites (George 1997). Spirorchiids are vascular system generalists, with a preference for the heart and arterial system of their turtle hosts (Platt & Brooks 1997). Adult parasites may cause endocarditis, arteritis and thrombosis of the blood vessels (Gordon et al. 1998). In addition to direct pathological effects induced by the adult parasite, eggs released within the vascular system may be transported to remote areas, such as the central nervous system (CNS), where they lodge in small vessels, often initiating a mild to severe granulomatous inflammatory response. The eggs can also migrate through blood vessel walls, causing tissue damage and inflammation in adjacent tissues (Gordon et al. 1998). Spirorchiid parasitism may promote secondary gram-negative bacterial infections (Raidal et al. 1998). Meningitis and encephalitis have been reported in green...

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Section: Methodsmentioning

confidence: 99%

Neurological disease in wild loggerhead sea turtles Caretta caretta

Jacobson

Homer²,

Stacy³

et al. 2006

Dis. Aquat. Org.

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“…This method is under increasing criticism not only from an ethical point of view but also because of its limited sensitivity and inability to distinguish between toxins of different groups (Quilliam and Wright, 1995;Hess et al, 2006). A number of alternative methods have been proposed ranging from cytotoxicity assays (Aune et al, 1991;Amzil et al, 1992.;Tubaro et al, 1996a), protein phosphatase assays (Luu et al, 1993;Honkanen et al, 1996;Simon and Vernoux, 1994;Tubaro et al, 1996b;Isobe et al, 1995;Vieytes et al, 1997), liquid chromatography (LC) coupled with fluorescence detection and LC coupled with mass spectrometry, capillary electrophoresis and numerous immunosorbent methods (James et al, 2000). Some of these methods lack the required sensitivity and others require long sample preparation.…”

Section: Introductionmentioning

confidence: 99%

Development of a monoclonal antibody binding okadaic acid and dinophysistoxins-1, -2 in proportion to their toxicity equivalence factors

Stewart

Elliott

Walker³

et al. 2009

Toxicon

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a b s t r a c tOkadaic acid (OA) and structurally related toxins dinophysistoxin-1 (DTX-1), and DTX-2, are lipophilic marine biotoxins. The current reference method for the analysis of these toxins is the mouse bioassay (MBA). This method is under increasing criticism both from an ethical point of view and because of its limited sensitivity and specificity. Alternative replacement methods must be rapid, robust, cost effective, specific and sensitive. Although published immuno-based detection techniques have good sensitivities, they are restricted in their use because of their inability to: (i) detect all of the OA toxins that contribute to contamination; and (ii) factor in the relative toxicities of each contaminant. Monoclonal antibodies (MAbs) were produced to OA and an automated biosensor screening assay developed and compared with ELISA techniques. The screening assay was designed to increase the probability of identifying a MAb capable of detecting all OA toxins. The result was the generation of a unique MAb which not only cross-reacted with both DTX-1 and DTX-2 but had a cross-reactivity profile in buffer that reflected exactly the intrinsic toxic potency of the OA group of toxins. Preliminary matrix studies reflected these results. This antibody is an excellent candidate for the development of a range of functional immunochemical-based detection assays for this group of toxins.

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“…Testes de toxicidade com camundongos são eficientes, porém não identificam presença de variantes das toxinas, ou se outras toxinas estão sendo produzidas (AUNE; BERG, 1986), bem como os métodos com sacrifício de animais são cada vez mais desaconselhados para uso de rotina. Testes enzimáticos e imunoenzimáticos apresentam vantagens relacionadas a um custo mais baixo e não necessitam de profissionais com alto nível de especialização para realizar os testes RIVASSEAU et al, 1999;TUBARO et al, 1996). Contudo, esses testes enzimáticos também não identificam variantes das toxinas, e não detectam outras toxinas presentes, que não inibam a enzima ou para as quais não exista anticorpo.…”

Section: Realidades E Perspectivas Da Detecção De Toxinasunclassified

Toxins of cyanobacteria and microalgae: a challenge for aquatic ecotoxicology

Oliveira¹,

Filho²,

Bastos³

et al. 2010

Boletim Do Observatório Ambiental Alberto Ribeiro Lamego

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ResumoO objetivo desta revisão é de chamar a atenção para o crescente número de metabólitos secundários produzidos por microrganismos em ambientes aquáticos. Em especial pelas cianobactérias e pelas microalgas marinhas. O primeiro grupo é produtor de toxinas que têm por alvo o fígado, os nervos e a pele. As cianobactérias são as maiores responsáveis por eventos de intoxicação em ambientes aquáticos epicontinentais. Em ambientes marinhos, microalgas eucariotas como as diatomáceas e principalmente os dinoflagelados são produtores de ampla variedade de ficotoxinas. Ficotoxinas são causadoras de síndromes após consumo de mexilhão contaminado ou intoxicação ciguatérica por consumo de peixe (CFP). O quadro atual sinaliza atenção, pois muitas florações ocorrem sem notificação, o que impede importantes estudos sobre a história de florações sazonais. Aspectos metodológicos e de mão de obra qualificada também dificultam diagnósticos mais precisos sobre as florações. A cooperação entre profissionais com distintas formações como taxonomistas, químicos, biólogos e engenheiros ambientais é a base para estudos ecotoxicológicos que possam avaliar o risco destas florações e sugerir adaptações na legislação ambiental.Palavras-chave: Toxinas. Cianobactéria. Algas nocivas. AbstractThe purpose of this review is to draw attention to the growing number of secondary metabolites produced by microorganisms in aquatic environments, specially cyanobacteria and marine microalgae. The first group is a producer of major hepatotoxins, neurotoxins and dermatotoxins. Cyanobacteria are responsible for most

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A protein phosphatase 2A inhibition assay for a fast and sensitive assessment of okadaic acid contamination in mussels

Cited by 138 publications

References 14 publications

Neurological disease in wild loggerhead sea turtles Caretta caretta

Neurological disease in wild loggerhead sea turtles Caretta caretta

Development of a monoclonal antibody binding okadaic acid and dinophysistoxins-1, -2 in proportion to their toxicity equivalence factors

Toxins of cyanobacteria and microalgae: a challenge for aquatic ecotoxicology

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