Ballast water exchange was measured for the first time in Bilbao Harbour, one of the most active of northern Spain. Between 1997 and 2006, 41,900,980.34 ballast water tn were loaded and 13,272,709 tn were discharged. Bilbao Harbour appears to be mainly a source of ballast water, 90% of which would be discharged in European harbours. We estimated that vessels carrying liquid and solid bulk have higher probabilities of exporting ballast water, whereas those with liquid bulk and containers are more likely to introduce it. From 30 potentially harmful phytoplankton species identified to date near harbour facilities, there would be a high risk of exporting at least Alexandrium minutum, Dinophysis sp., Heterosigma akashiwo, Karlodinium sp., Ostreopsis cf. siamensis, Pfiesteria-like and Prorocentrum minimum. Invasion risk by ballast water was tested by analyzing the response of six strains of H. akashiwo from different geographic areas to varying salinity. Results show that successful growth of foreign strains would be possible.

Testing phytoplankton viability within ballast tanks and receiving waters of ballast water discharge remain understudied. Potentially harmful dinoflagellates and diatoms are transported via ballast water to Galveston Bay, Texas (USA), home to three major ports: Houston, Texas City and Galveston. Ballast water from vessels transiting the North Atlantic Ocean was inoculated into treatments representing low and high salinity conditions similar to the Ports of Houston and Galveston respectively. Phytoplankton in ballast water growout experiments were deemed viable and showed growth in low and mid salinities with nutrient enrichment. Molecular methods identified several genera: Dinophysis, Gymnodinium, Gyrodinium, Heterocapsa, Peridinium, Scrippsiella, Chaetoceros and Nitzschia. These phytoplankton genera were previously identified in Galveston Bay except Scrippsiella. Phytoplankton, including those capable of forming harmful algal blooms leading to fish and shellfish kills, are transported to Galveston Bay via ballast water, and are viable when introduced to similar salinity conditions found in Galveston Bay ports.

Continuous monitoring and early warning together represent an important mitigation strategy for harmful algal blooms (HAB). The coast of Texas experiences periodic blooms of three HAB dinoflagellates: Karenia brevis, Dinophysis ovum, and Prorocentrum texanum. A plankton image data set acquired by an Imaging FlowCytobot over a decade of operation was used to train and evaluate two new automated image classifiers. A 112 class, random forest classifier (RF_112) and a 112 class, convolutional neural network classifier (CNN_112) were developed and compared with an existing, 54 class, random forest classifier (RF_54) already in use as an early warning notification system. Both 112 class classifiers exhibited improved performance over the RF_54 classifier when tested on three different HAB species with the CNN_112 classifier producing fewer false positives and false negatives in most of the cases tested. For K. brevis and P. texanum, the current threshold of 2 cells.mL⁻¹ was identified as the best threshold to minimize the number of false positives and false negatives. For D. ovum, a threshold of 1 cell.mL⁻¹ was found to produce the best results with regard to the number of false positives/negatives. A lower threshold will result in earlier notification of an increase in cell concentration and will provide state health managers with increased lead time to prepare for an impending HAB.

Harmful or nuisance algal blooms can cause economic damage to fisheries and tourism. Additionally, toxins produced by harmful algae and ingested via contaminated shellfish can be potentially fatal to humans. The seas around the Orkney Islands, UK currently hold a number of toxic algal species which cause shellfishery closures in most years. Extensive and costly monitoring programs are carried out to detect harmful microalgae before they reach action levels. However, the ability to distinguish between toxic and non-toxic strains of some algae is not possible using these methods. The microarrays for the detection of toxic algae (MIDTAL) microarray contains rRNA probes for toxic algal species/strains which have been adapted and optimized for microarray use. In order to investigate the use of the chip for monitoring in the Orkney Islands, samples were collected between 2009 and 2011 from Brings Deep, Scapa Flow, Orkney Islands, UK; RNA was extracted and hybridized with generation 2 and 3.1 of the chip. The data were then compared to cell counts performed under light microscopy and in the case of Alexandrium tamarense to qPCR data targeting the saxitoxin gene and the LSU-rRNA gene. A good agreement between cell numbers and microarray signal was found for A. tamarense, Pseudo-nitzschia sp., Dinophysis sp. (r < 0.5, for all) in addition to this there the chip successfully detected a large bloom of Karenia mikimotoi (r < 0.70) in August and September 2011. Overall, there was good improvement in probe signal between generation 2 and generation 3.1 of the chip with much less variability and more consistent results and better correlation between the probes. The chip performed well for A. tamarense group I signal to cell numbers in calibrations (r > 0.9). However, in field samples, this correlation was slightly lower suggesting interactions between all species in the sample may affect signal. Overall, the chip showed it could identify the presence of target species in field samples although some work is needed to improve the quantitative nature of the chip before it would be suitable for monitoring in the Orkney Islands.

We reported previously that okadaic acid (OA) and dinophysistoxin-1 (DTX1) were responsible for diarrhetic shellfish poisoning (DSP) incidents due to consuming cultivated mussels (Mytilus galloprovincialis) in coastal cities near the East China Sea in May 2011. Pectenotoxin-2 (PTX2) and its seco acids were also present in these mussels. Causative species of microalgae were not identified because detailed information on the location of the contaminated shellfish was not recorded. In order to explore potential causes for these poisoning events, the lipophilic toxin profiles in picked cells of Dinophysis and in shellfish samples collected from two mariculture zones in the East China Sea were analyzed in the present study. Single-cell isolates (100 cells total for each location) of Dinophysis were collected from the aquaculture zones of Gouqi Island (Ningbo City, Zhejiang Province) and Qingchuan Bay (Ningde City, Fujian Province) in July and September 2013, respectively, for lipophilic toxin profiling. Shellfish samples collected over the course of a year from the Gouqi Island aquaculture zone and mussels (M. galloprovincialis) collected four times from the Qingchuan Bay aquaculture zone were tested for lipophilic toxins by LC-MS/MS. The Dinophysis cells isolated from both sampling sites were identified under the light microscope as Dinophysis caudata. Average quota of PTX2, the predominant toxin in D. caudata isolated from the coastal waters of Gouqi Island and Qingchuan Bay, was 0.58 and 2.8 pg/cell, respectively. Only trace amounts of OA and DTX1 were detected in D. caudata. PTX2, PTX2sa, 7-epi-PTX2sa, OA, and/or DTX1 were found in samples of mussels (M. galloprovincialis and Mytilus coruscus) collected in the Gouqi Island aquaculture zone from the end of May to the beginning of July 2013. PTX2, PTX2sa, and 7-epi-PTX2sa were also detected in oyster (Crassostrea gigas) during that period, but almost no OA and DTX1 were present. Gymnodimine (GYM) was detected in almost all mussel (M. coruscus) samples, with the highest levels occurring in winter. Trace amounts of pectenotoxins (PTXs) and OAs were also found in mussels (M. galloprovincialis) collected from Qingchuan Bay. D. caudata is suggested as an important source of PTXs in shellfish cultivated in the East China Sea. This is the first report of toxin profiles for single-cell isolates of Dinophysis in the East China Sea.