Germination of cysts serves as inoculum for the proliferation of some dinoflagellates, and cyst abundance in sediments represents crucial information to understand and possibly predict Harmful Algae Blooms (HABs). Cyst distribution is related to the physical characteristics of the sediments and the hydrodynamics (circulation) of a particular region. In the northern Gulf of California (nGC) several Gymnodinium catenatum HABs have been recorded. However, the presence of resting cysts and the effect of hydrodynamics on their distribution in the nGC have not been investigated. This study evaluated cyst abundance, distribution and their relation to local circulation in surface sediments during two periods that coincided with a non-bloom year condition (July 2016) and after a major HAB registered in the nGC that occurred in January 2017. Also, a numerical ocean model was implemented to characterize the transport and relocation of cysts and sediments in the region. Gymnodinium catenatum cysts were heterogeneously distributed with some areas of high accumulation (as high as 158 cyst g−1, and 27% of total cyst registered). Cysts seemed to be transported in an eastward direction after deposition and accumulated in an extensive area that probably is the seedbed responsible for the initiation of HABs in the region. The nGC is a retention area of cysts (and sediments) that permit the formation of seedbeds that could be important for G. catenatum HAB development. Our results provide key information to understand G. catenatum ecology and specifically, to understand the geographic and temporal appearance of HABs in the nGC.

Contamination of shellfish by paralytic shellfish poisoning (PSP) toxins poses an economic threat to shellfish farmers. As contaminated shellfish cannot be harvested for long periods of time, it would be very useful to develop processes to optimise and shorten their detoxification. In this study, Pacific oysters Crassostrea gigas were first experimentally contaminated over a period of 13 days with a continuous flow of toxic Alexandrium minutum cultures at concentrations ranging from 150 to 200 cell ml-1 (toxin content after 13 days of contamination 438 $\mu $ g STX equiv. 100 g−1 wet weight). Then, two different detoxification treatments were tested and showed detoxification rates greater than those observed in coastal environments. The first treatment consisted of feeding oysters on Skeletonema costatum, at a concentration of 2000 cell ml−1 to speed up detoxification rates. The second detoxification method used the same Skeletonema costatum diet, supplemented with silt particles at a concentration of 20 mg L−1. A control was also set up by placing contaminated oysters in seawater with no additional algal food. The detoxification experiment lasted 8 days. Toxin contents were analysed by liquid chromatography with fluorescence detection (LC-FD). The S. costatum diet significantly reduced the time needed for oysters to reach the sanitary threshold (80 $\mu $ g STX equiv. 100 g−1 wet weight), but no effect of the silt supplement could be demonstrated conclusively. These different detoxification methods did not influence toxin biotransformations as observed in oyster tissues, i.e. epimerisation and decarbamoylation of gonyautoxins 2 and 3.

The impact of toxic outbreaks of Gymnodinium catenatum, producer of paralytic shellfish poisoning toxins (PSP toxins) and Pseudonitzschia spp., producer of amnesic shellfish poisoning toxins (ASP toxins), was studied throughout the culture cycle of the king scallop Pecten maximus (length from 40 to 100 mm). Toxin concentration, in plankton and in the digestive gland of the scallops, other variables that describe the state of the cultured scallops (mortality, growth, length/weight ratio, gonadosomatic index) and the main environmental conditions (temperature and in vivo fluorescence) were determined from December 2001 to April 2003. Temperature ranged from 14 °C during the winter to 22 °C at the end of July. In vivo fluorescence ranged from not detectable to 25 relative units, with a maximum also recorded at the end of July. ASP toxin levels, in both plankton and organisms, were high during the first half of the sampling period (maximum 450 ng domoic acid L−1, in plankton, and 175 µg·g−1 of scallop digestive gland) when the scallops were juvenile. PSP toxins attained larger concentrations in the second half of the sampling period (maximum of 1.8 nmol L−1 in plankton and 550 nmol g−1 of digestive gland, of gonyautoxins after hydrolysis). Several mortality events took place during the period studied, but none of them coincided neither with an ASP or PSP outbreak nor with long exposure of the scallops to these toxins. One of the recorded mortalities occurred simultaneously with a high concentration of PSP in the scallops, but also with an important spawning. Additionally, it was not coincidental with the bloom of toxic plankton, thus, post-spawning stress seemed to be the main responsible for the mortality. No appreciable effect of ASP or PSP toxinson growth or reproductive state of the cultured scallops was found. Therefore, neither ASP nor PSP toxins appeared to represent a direct threat to the culture of Pecten maximus in that case.

A simple system was used to simulate the effect of alternating toxic (paralytic shellfish poisoning toxins) and non-toxic microalgal diets on oyster feeding behaviors and rates of toxin accumulation. These experimental conditions were meant to reflect, to some extent, the incoming and outgoing fluxes of toxic algae observed at the mouth of the Penzé estuary (Northern Brittany, France). Physiological and toxicological parameters were estimated based on fluorescence measurements recorded continuously at the outlet of each experimental tank, which contained a single oyster. Q tox, this variable describes toxin uptake in oysters, it was used (instead of the toxin ingestion rate): i) in simple graphical analyses, ii) as well as in one- and two-compartment models. Results show that toxin uptake varies widely from one individual to another and is not proportional to the concentration of toxic algae in sea water. A one-compartment model with individual fluorescence recordings as “input” data gave questionable results, however, a two-compartment model was found to effectively describe contamination kinetics in oysters. Limitations of this model as well as possible improvements are discussed.

Several experiments using a self-regulated system were conducted to define the factors likely to influence the uptake of paralytic shellfish poison (PSP) by oysters in the Penzé estuary (France, Brittany). Each 4-day experiment was carried out in a recirculated sea water system using 15 Pacific oysters (Crassostrea gigas) separated from each other and supplied with unfiltered natural seawater containing alternatively toxic (Alexandrium minutum) or non-toxic (Skeletonema costatum) algal diets. The food supply and exposure times to toxic diets were determined according to field studies of the upstream and downstream movement of patches containing A. minutum. The experimental parameters corresponded roughly to the hydrological conditions generally observed in June when tidal coefficients are lowest and blooms occur: (i) A. minutum concentrations in sea water of 200, 5000 and 10 000 cell ml−1; (ii) inorganic matter consisting of 5 and 15 mg L−1 of calcinated muddy sediments; and (iii) low and high tide salinities of 25 and 35‰, respectively. Significant experimental contamination (greater than the 80 µg STX equiv. 100 g-1 sanitary threshold) occurred after 4 days of exposure for the monospecific A. minutum diet (20-200 cell ml−1) and alternated A. minutum and S. costatum diets (5000 and 20 000 cell ml−1, respectively). Contamination levels were less than the sanitary threshold for alternated A. minutum/S. costatum diets of 200 and 20 000 cell ml−1, respectively, and for a monospecific A. minutum diet (1000−10 000 cell ml−1). In the last case, the accumulation rate was quite low, possibly because of inhibition of the filtration rate related to a lower biodeposit production rate and decreased feeding time activity. The addition of inorganic matter appeared to play a significant role in the observed increase of toxin uptake, whereas salinity was not a determining factor for toxin accumulation rates. These last observations were corroborated by statistical analysis and stepwise multiple linear regressions integrating all or some of the experimental parameters.

There is increasing evidence that bacterial–algal interactions play a role in Harmful Algal Bloom (HAB) ecology. Bacteria that are associated with bloom-forming algal species, specifically toxic dinoflagellate algae, have been implicated in the production and biotransformation of paralytic shellfish toxins (PSTs). To clarify the role that these bacteria may play in the production of PSTs, it is desirable to identify and localize the bacteria associated with the dinoflagellates and enumerate them during the course of the algal blooms that the toxic dinoflagellates produce. Because 16S rRNA-targeted probes offer the possibility of both, we previously made and tested probes for some putatively toxigenic bacteria isolated from cultures of the PSP-related dinoflagellates Alexandrium tamarense, A. affine and A. lusitanicum. The bacteria isolated from the dinoflagellates belong primarily to the alpha-proteobacterial group of Roseobacter and the gamma-proteobacterial group of Alteromonas. Here, we report the successful application of these probes to Lugol's-fixed seawater samples. We detected these bacteria in high numbers in the water column when Alexandrium spp. were both present and absent, and during periods when mussels contained PSTs.

The effect of N- and P-limitation on the growth and paralytic shellfish toxin content of Alexandrium fundyense was studied in a series of batch cultures in which the N[ratio ]P supply mass ratio was varied from less than 1 up to 160 (cf. Redfield ≡ 7·2). N- and P-limitation of growth was observed at the lower and higher ratios respectively, with dual limitation between. Cellular parameters were similar during the exponential phase of growth, regardless of external N[ratio ]P ratio. Upon nutrient exhaustion cell yields and internal pools were affected, although these also varied according to the N-source (nitrate vs. ammonium). Generally N-limitation alone led to a higher C[ratio ]N ratio, lower C[ratio ]P and N[ratio ]P ratios and a decreased toxin content in stationary-phase cells. P-limitation led to increased cell size and cell quotas, lower C[ratio ]N ratios, and higher C[ratio ]P and N[ratio ]P ratios (to maxima of 350 and 35 respectively). Complex internal P-fractionation schemes were found unnecessary to determine the P-status of cells and extraction using potassium persulphate, cold water and cold HCl alone were adequate. Severe P-limitation led to increased toxin content but only when N was also limiting, suggesting a synergistic effect of N and P availability on toxin synthesis and turnover. A positive relationship was found between toxin content and the intracellular concentration of arginine, but this varied with the nutrient status (in particular P-status) of the cells.