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Mardones Find articles by Jorge I.

Hallegraeff Find articles by Gustaaf M. E-Tech KMWL01 authors have declared that no competing interests exist. Conceived and designed the experiments: Received Feb 20; Accepted Jun This article has been cited by other articles in PMC. Associated Data All relevant data are within the paper. Abstract Quantification of the role of reactive oxygen species, phycotoxins and fatty acids in fish toxicity by harmful marine microalgae remains inconclusive. An in vitro fish gill from rainbow trout Oncorhynchus mykiss assay was used to simultaneously assess the effect in superoxide dismutase, catalase and lactate dehydrogenase enzymatic activities caused by seven species of ichthyotoxic microalgae Chattonella marina, Fibrocapsa japonica, Heterosigma akashiwo, Karenia mikimotoi, Alexandrium catenella, Karlodinium veneficum, Prymnesium parvum.

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Quantification of superoxide production by these algae was also performed. The effect of purified phycotoxins E-Tech KMWL01 crude extracts was compared, and the effect of fatty acids is discussed.

E-Tech KMWL01 raphidophyte Heterosigma and dinoflagellate Alexandrium were the least toxic and had low superoxide production, except when A. Catalase showed no changes in activity in all the treatments. A strong relationship between gill cell viability and superoxide production or superoxide dismutase was not observed. These and our previous findings involving the role of fatty acids confirm that superoxide radicals are only partially involved in ichthyotoxicity and point to a highly variable contribution by other compounds such as lipid peroxidation products e.

Introduction The multi-million dollar economic impacts from harmful microalgae on fish farming have been well documented and far E-Tech KMWL01 the comparable impacts on shellfish industries, where the main impact is direct loss of the product due to mass mortalities [ 1 — 5 ]. For instance, E-Tech KMWL01 economic losses in Japan between and were The species that have created highest impacts on fish farms worldwide are the unarmoured Cochlodinium polykrikoides, Karenia mikimotoi dinoflagellatesChattonella marina, C.


Despite the impacts of these events, it remains to be clarified how E-Tech KMWL01 microalgae that do not produce chemically characterized toxins different from neurotoxic, diarrhetic, amnesic and paralytic shellfish poisoning causing metabolites are killing finfish. Ichthyotoxicity has been attributed variously to production of free fatty acids [ 8 — 10 ] and reactive oxygen species or ROS the superoxide radical O2 - in particular [ 1112 ] and occasionally to chemically defined phycotoxins such as brevetoxins or karlotoxins [ 13 — 15 ].

However, not all ichthyotoxic microalgae produce these compounds in amounts that can account for their impacts on fish. Reactive oxygen species are the result of electron transport, as occurs in metabolic processes within the cell. Reactive E-Tech KMWL01 species are produced during respiration and photosynthesis, and can be significantly reduced using photosynthesis E-Tech KMWL01, which suggests that fish mortality may be more prominent during day light hours [ 17 ]. Sensitive assays have been developed to quantify superoxide and hydroxyl radicals in aqueous solution.

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Superoxide lifetime in seawater has been measured in the range of 10— s [ 19 ]. The improved MCLA 2-methyl 4-methoxyphenyl -3,7-dihydroimidazo[1,2-a]pyrazin-3 7H -one assay adapted by E-Tech KMWL01 et al.

This assay is based on the chemilumninescence of MCLA generated when it reacts with the superoxide radicals in the medium, as being produced by E-Tech KMWL01 algae. The signal is measured by a luminescence detector using a microplate reader. Screening for generation of superoxide by a wide range of microalgae, has conclusively shown that raphidophytes of the genus E-Tech KMWL01 are the greatest O2 - producers, generating up to 18 times more superoxide than other ichthyotoxic species, including raphidophytes and dinoflagellates [ 21 ].


There exists controversy in the ichthyotoxic mechanism of the dinoflagellate Cochlodinium polykrikoides, since early studies suggested the role of ROS but others found only trace levels, concluding that C. Ichthyotoxic unarmoured microalgae are very fragile and susceptible to cell rupture; when this occurs, a E-Tech KMWL01 of reactive compounds are released into the water. These compounds affect the fish mainly via gill damage during respiration, and this damage can be accelerated with hyperventilation under continuing stress conditions [ 12 ].

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With some species, such as the armoured dinoflagellate Karlodinium, cell lysis has been conclusively demonstrated to be critical for ichthyotoxicity [ 26 ], and similarly for the naked flagellate Chattonella more fragile strains are more potent fish killers [ 27 ]. However, when ROS production exceeds these defenses, organisms can undergo oxidative stress through damage to proteins, DNA and lipids, creating physiological changes that may lead to death. Similarly, in pioneering fish challenge experiments by Yang et al. In our previous experimental studies we presented a sensitive in vitro assay to test toxicity of harmful phytoplankton using a gill cell line from rainbow trout Oncorhynchus mykiss as a model [ 2730 ].

The premise of this E-Tech KMWL01 is that fish gill damage is the first line of attack in harmful microalgae killing fish [ 31 ].


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