Unprecedented Alexandrium blooms in a previously low biotoxin risk area of Tasmania, Australia.

During October 2012, a shipment of blue mussels (Mytilus galloprovincialis) from the poorly monitored east coast of Tasmania, Australia, was tested by Japanese import authorities and found to be contaminated with unacceptable levels of Paralytic Shellfish Toxins (PSTs; 10 mg/kg). Subsequently local oysters, scallops, clams, the viscera of abalone and rock lobsters were also found to be contaminated. This led to a global product recall and loss to the local economy of AUD 23M. Following low toxicity during 2013 and 2014 and implementation of minimal shellfish farm closures, a more severe bloom event occurred during July-November 2015 and again June-September 2016 (up to 300,000 Alexandrium cells/L; 24 mg/kg PST in mussels, 6 mg/kg in Crassostrea gigas oysters), also causing 4 human illnesses resulting in hospitalization after consumption of wild shellfish. While Alexandrium tamarense had been detected in low concentrations in southeastern Australia since 1987, all cultured strains belonged to the mostly non-toxic group 5 (now designated A. australiense; detected since 1987) and weakly toxic group 4 (A. pacificum; detected in 1997). In contrast, the 2012 to 2016 outbreaks were dominated by highly toxic group 1 (A. fundyense) never detected previously in the Australian region. Molecular analyses suggest that A. fundyense may have been a cryptic ribotype previously present in Tasmania, but newly stimulated by altered water column stratification conditions driven by changing rainfall and temperature patterns. Increased seafood and plankton monitoring of the area now include the implementation of Alexandrium qPCR, routine Neogen™ immunological and HPLC PST tests, but ultimately may also drive change in harvesting strategies and aquaculture species selection by the local seafood industry

Field validation of the southern rock lobster paralytic shellfish toxin monitoring program in Tasmania, Australia

Paralytic shellfish toxins (PST) are found in the hepatopancreas of Southern Rock Lobster Jasus edwardsii from the east coast of Tasmania in association with blooms of the toxic dinoflagellate Alexandrium catenella. Tasmania’s rock lobster fishery is one of the state’s most important wild capture fisheries, supporting a significant commercial industry (AUD 97M) and recreational fishing sector. A comprehensive 8 years of field data collected across multiple sites has allowed continued improvements to the risk management program protecting public health and market access for the Tasmanian lobster fishery. High variability was seen in toxin levels between individuals, sites, months, and years. The highest risk sites were those on the central east coast, with July to January identified as the most at-risk months. Relatively high uptake rates were observed (exponential rate of 2% per day), similar to filter-feeding mussels, and meant that lobster accumulated toxins quickly. Similarly, lobsters were relatively fast detoxifiers, losing up to 3% PST per day, following bloom demise. Mussel sentinel lines were effective in indicating a risk of elevated PST in lobster hepatopancreas, with annual baseline monitoring costing approximately 0.06% of the industry value. In addition, it was determined that if the mean hepatopancreas PST levels in five individual lobsters from a site were −1, there is a 97.5% probability that any lobster from that site would be below the bivalve maximum level of 0.8 mg STX equiv. kg−1. The combination of using a sentinel species to identify risk areas and sampling five individual lobsters at a particular site, provides a cost-effective strategy for managing PST risk in the Tasmanian commercial lobster fishery

Multiscale-Multifractal Assessment of Heart Rate Variability in Shift Workers by Detrended Fluctuation Analysis

Alteration of the circadian rhythm of the sleep-wake cycle is observed in shift workers, causing sleep disorders and changes in the cardiac autonomic nervous system. The multifractal-multiscale structure of inter-beat intervals (IBI) during sleep was analyzed in healthy shift working females by the multiscale-multifractal detrended fluctuation analysis (MMF-DFA) method. The MMF-DFA was applied to estimate the self-similarity coefficients α(q, τ) considering moment orders q between - 5 and + 5, and scales τ between 8 and 150 s. During daytime sleep, for q> 2, the α(q, τ) coefficients were very similar to those of nighttime sleep along τ. However, significantly higher scaling coefficients were observed during daytime sleep than during nighttime sleep, at small scales 44 ≤ τ≤ 68 for 1 ≤ q≤ 2, and at large scales 219 ≤ τ≤ 414 for negative moment orders q. The results suggest an alteration in the autonomic nervous system of shift workers, which could increase the risk of cardiovascular disease. Also, multifractal surface assessment of scale coefficients during sleep could be a tool to complement and improve the assessment of HRV alterations due to shift work

Unraveling the Karenia selliformis complex with the description of a non-gymnodimine producing Patagonian phylotype

Karenia selliformis is a bloom-forming toxic dinoflagellate known for production of gymnodimines (GYMs) and causing mass mortalities of marine fauna. Blooms have been reported from coastal waters of New Zealand, Mexico, Tunisia, Kuwait, Iran, China and Chile. Based on molecular phylogeny, morphology, toxin production, pigment composition and cell growth of Chilean K. selliformis isolated in 2018 (CREAN_KS01 and CREAN_KS02), this study revealed a more complex diversity within this species than previously thought. A phylogenetic reconstruction based on the large sub-unit ribosomal nucleotide (LSU rDNA) and Internal Transcriber Spacer (ITS) sequences of 12 worldwide isolates showed that within the K. selliformis clade there are at least two different phylotypes with clear phenotypic differences. Morphological differences related to the dorsal-ventral compression, shape of the hyposome and the presence of pores on the left lobe of the hyposome. A comparison of pigment signatures among worldwide isolates revealed the existence of both acyl-oxyfucoxanthin and fucoxanthin-rich strains within the phylotypes. A LC-MS/MS screening on both Chilean 2018 K. selliformis strains showed for first time no GYMs production among cultured clones of this species. However, both CREAN_KS01 and CREAN_KS02 contained two compounds with the same mass transition as brevenal, a brevetoxin related compound. A fish gill cell-based assay showed that the CREAN_KS02 strain was highly cytotoxic but pure GYM standard did not exhibit loss of cell viability, even at cell concentrations equivalent or exceeding those reported in nature. The fatty acid profile of CREAN_KS02 included high levels of saturated (14:0; 16:0) and polyunsaturated (18:3ω6+18:5ω3; 22:6ω3) fatty acids but superoxide production in this strain was low (0.86±0.53 pmol O2− cell−1 h−1). A factorial T-S growth experiment using the CREAN_KS02 strain showed a μmax of 0.41±0.03 d−1 at high salinity and temperature, which points to its optimal environmental niche in offshore waters during the summer season. In conclusion, the present study provides evidence for significant genetic and phenotypic variability among worldwide isolates, which points to the existence of a K. selliformis “species complex”. The massive fauna mortality during K. selliformis bloom events in the Chilean coast cannot be explained by GYMs nor brevetoxins, but can to a large extent be accounted for by the high production of long-chain PUFAs and/or still uncharacterized highly toxic compounds

Progress in Understanding Algal Bloom-Mediated Fish Kills: The Role of Superoxide Radicals, Phycotoxins and Fatty Acids

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). 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 raphidophyte Chattonella was the most ichthyotoxic (gill cell viability down to 35%) and also the major producer of superoxide radicals (14 pmol cell-1 hr-1) especially after cell lysis. The raphidophyte Heterosigma and dinoflagellate Alexandrium were the least toxic and had low superoxide production, except when A. catenella was lysed (5.6 pmol cell-1 hr-1). Catalase showed no changes in activity in all the treatments. Superoxide dismutase (SOD) and lactate dehydrogenase exhibited significant activity increases of ≤23% and 51.2% TCC (total cellular content), respectively, after exposure to C. marina, but SOD showed insignificant changes with remaining algal species. A strong relationship between gill cell viability and superoxide production or superoxide dismutase was not observed. Purified brevetoxins PbTx-2 and -3 (from Karenia brevis, LC50 of 22.1 versus 35.2 μg mL-1) and karlotoxin KmTx-2 (from Karlodinium; LC50 = 380 ng mL-1) could almost entirely account for the fish killing activity by those two dinoflagellates. However, the paralytic shellfish toxins (PST) GTX1&4, C1&C2, and STX did not account for Alexandrium ichthyotoxicity. Only aqueous extracts of Alexandrium were cytotoxic (≤65% decrease of viability), whereas crude methanol and acetone extracts of Chattonella, Fibrocapsa, Heterosigma, Karlodinium and Prymnesium decreased cell viability down to 0%. 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.g. aldehydes)