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Mardones Find articles by Jorge I. Hallegraeff Find articles by Gustaaf M. The authors have declared that no competing interests exist. Conceived E-Tech KMWL01 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 E-Tech KMWL01.
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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 E-Tech KMWL01 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. Quantification of superoxide production by these algae was also performed. The effect of purified phycotoxins and crude extracts was compared, and the effect of fatty acids is discussed.
The E-Tech KMWL01 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 E-Tech KMWL01 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.
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E-Tech KMWL01 Introduction The multi-million dollar economic impacts from harmful microalgae on fish farming have been well E-Tech KMWL01 and far exceed the comparable impacts on shellfish industries, where the main impact is direct loss of the product due to mass mortalities [ 1 — 5 ]. For instance, the 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 these 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.
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Reactive oxygen species are produced during respiration and photosynthesis, and can be significantly reduced using photosynthesis blockers, 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 E-Tech KMWL01. 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 Godrant 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 the algae. The signal is measured by a luminescence detector using a microplate reader.
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Screening for generation of superoxide E-Tech KMWL01 a wide range of microalgae, has conclusively shown that raphidophytes of the genus Chattonella 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 E-Tech KMWL01 are very fragile and susceptible to cell rupture; when this occurs, a cocktail 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 ]. 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 E-Tech KMWL01 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 E-Tech KMWL01 Oncorhynchus mykiss as a model [ 2730 ].
The premise of this approach is that fish gill damage is the first line of attack in harmful microalgae killing fish [ 31 ]. In the present study, we challenged the gill cells against different harmful microalgae and measure the activity E-Tech KMWL01 antioxidant enzymes in the gill cells, as well as superoxide production by the microalgae. The effect of purified phycotoxins and crude algal extracts was also assessed in gill cells to resolve the role of superoxide radicals, phycotoxins or other algal compounds in ichthyotoxicity.
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