Ichthyotoxicity found in cultured media of Protogonyaulax spp.

Ichthyotoxicity was observed in the cultured media from Protogonyaulax catenella and P. tamarensis. No paralytic shellfish poison was detected in the cultured media. Test fish in the cultured media died showing signs of hypoxia. Histological observations of the gills of intoxicated fish indicated that the epithelial cells were swollen and exfoliated from the pillar cell. The cultured media had hemolytic activity against erythrocytes of various animals. The media also decreased the elasticity of eggs of rainbow trout Salmo gairdneri. From these results, it is concluded that the cultured media of both species of Protogonyaulax contain a cytotoxin which is lethal to fish.     

Presence of Alexandrium catenella and paralytic shellfish toxins in finfish, shellfish and rock crabs in Monterey Bay, California, USA

The central California coast is a highly productive, biodiverse region that is frequently affected by the toxin-producing dinoflagellate Alexandrium catenella. Despite the consistent presence of A. catenella along our coast, very little is known about the movement of its toxins through local marine food webs. In the present study, we investigated 13 species of commercial finfish and rock crabs harvested in Monterey Bay, California for the presence of paralytic shellfish toxins (PSTs) and compared them to the presence of A. catenella and PSTs in sentinel shellfish over a 3-year period. Between 2003 and 2005, A. catenella was noted in 55% of surface water samples (n = 307) and reached a maximum concentration of 17,387 cells L⁻¹ at our nearshore site in Monterey Bay. Peak cell densities occurred in the month of July and were associated with elevated shellfish toxicity in the summers of 2004 and 2005. When A. catenella was present, particulate PSTs were detected 71% of the time and reached a maximum concentration of 962 ng STXeq L⁻¹. Of the 13 species tested, we frequently detected PSTs in Pacific sardines (Sardinops sagax; maximum 250 μg STXeq 100 g⁻¹), northern anchovies (Engraulis mordax; maximum 23.2 μg STXeq 100 g⁻¹), brown rock crabs (Cancer antennarius; maximum 49.3 μg STXeq 100 g⁻¹) and red rock crabs (C. productus; 23.8 μg STXeq 100 g⁻¹). PSTs were also present in one sample of Pacific herring (Clupea pallas; 13.3 μg STXeq 100 g⁻¹) and one sample of English sole (Pleuronectes vetulus; 4.5 μg STXeq 100 g⁻¹), and not detected in seven other species of flatfish tested. The presence of PSTs in several of these organisms reveals that toxins produced by A. catenella are more prevalent in California food webs than previously thought and also indicates potential routes of toxin transfer to higher trophic levels. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Preying on commercial fisheries and accumulating paralytic shellfish toxins: a dietary analysis of invasive Dosidicus gigas (Cephalopoda Ommastrephidae) stranded in Pacific Canada

In fall of 2009, several mass strandings of Humboldt squid (Dosidicus gigas) occurred on Vancouver Island (49°7′60N 125°54′0W). Morphological dissections coupled with DNA barcoding of stomach contents revealed Sardinops sagax (Pacific sardine) and Clupea pallasii (Pacific herring) as their primary prey. Plastic nurdles, fishing line, bull kelp, eelgrass, and a guillemot feather were also discovered. The primary prey, Pacific sardines and Pacific herring, are known to bioaccumulate paralytic shellfish toxins (PSTs); additionally, both PSTs and domoic acid (DA) have been implicated in other mass strandings. Therefore, stomach contents, and other tissues when possible, were tested for PSTs and DA. Testing revealed DA concentrations below regulatory guidance levels for human consumption, yet PSTs were well in excess. Though we cannot conclude that PSTs were the definitive cause of the strandings, our findings are the first report of PSTs in D. gigas.     

Dynamics and physiology of saxitoxin production by the dinoflagellatesAlexandrium spp.

Toxin content (fmol cell^–1) and a suite of elemental and macromolecular variables were measured in batch cultures of the dinoflagellatesAlexandrium fundyense, A. tamarense andAlexandrium sp. from the southern New England region, USA. A different perspective was provided by semicontinuous cultures which revealed sustained, steady-state physiological adaptations by cells to N and P limitation. Two types of variability were investigated. In batch culture, changes in nutrient availability with time caused growth stage variability in toxin content, which often peaked in mid-exponential growth. A second type of variability that could be superimposed on growth stage differences is best exemplified by the high toxin content of cells grown at suboptimal temperatures. Calculations of the net rate of toxin production (R _tox ; fmol cell^–1 d^–1) for these different culture treatments and modes made it possible to separate the dynamics of toxin production from cell division. Over a wide range of growth rates, cells produced toxin at rates approximating those needed to replace losses to daughter cells during division. The exception to this direct proportionality was with P limitation, which was associated with a dramatic increase in the rate of toxin production as cells stopped dividing due to nutrient limitation in batch culture. Growth stage variability in batch culture thus reflects small imbalances (generally within a factor of two) between the specific rates of toxin production and cell division. N limitation and CO₂ depletion both affect pathways involved in toxin synthesis before those needed for cell division; P limitation does the opposite. The patterns of toxin accumulation were the same as for major cellular metabolites or elemental pools. The highest rates of toxin production appear to result from an increased availability of arginine (Arg) within the cell, due to either a lack of competition for this amino acid from pathways involved in cell division or to increased de novo synthesis. There were no significant changes in toxin content with either acclimated growth at elevated salinity, or with short term increases or decreases of salinity. These results demonstrate that toxin production is a complex process which, under some conditions, is closely coupled to growth rate; under other conditions, these processes are completely uncoupled. Explanations for the observed variability probably relate to pool sizes of important metabolites and to the differential response of key biochemical reactions to these pool sizes and to environmental conditions.     

Modelling of paralytic shellfish toxin biotransformations in the course of Crassostrea gigas detoxification kinetics

The aim of this study was to evaluate the importance of biotransformation of paralytic shellfish toxins during the detoxification process in contaminated oysters. Mathematical models based upon the detoxification patterns of digestive gland and other tissues were developed. It was demonstrated that biotransformations do not seem to play an important role in digestive gland or other tissue detoxification kinetics with our data set. Moreover, different toxin transfers from digestive gland toward other tissues were investigated. No significant transfer was highlighted in our data set. These first conclusions were drawn after comparing the results obtained from 13 biotransformations and identifiable transfer scenarios. Finally, to determine a more robust model, all 12 states corresponding to toxic compounds and tissues were aggregated into a single state model. The best adjustment was obtained with a simple one-compartment model based on total flesh toxicity with elimination rate expressed by a function depending on initial concentrations of GTX3 and GTX2 (i.e. the two major toxic compounds found in contaminated oysters).     

Effects of nutrient limitation on toxin production and composition in the marine dinoflagellate Protogonyaulax tamarensis

Toxin production was measured by high pressure liquid chromatography (HPLC) when the marine dinoflagellate Protogonyaulax tamarensis (NEPCC 255) was grown under nitrogen or phosphorus limitation. The major toxins found in P. tamarensis (255) consisted of (N21-SO ₃ ^- )STX (11%), (N21-SO ₃ ^- )NeoSTX (44%), and [(N21-SO ₃ ^- )GTX₂ plus (N21-SO ₃ ^- )GTX₃] (20%). Total toxin content on a per cell basis was high for cultures in log phase (30 to 40 fmol cell^-1) and then decreased to ca 20 fmol cell^-1 as the cultures entered stationary phase. There was a gradual decrease in the toxin content per cell during nitrogen-limited stationary phase to ca 3 fmol cell^-1 or less. Phosphorus-limited cultures showed a markedly different response than nitrogen-limited cultures. Toxin content in P-limited cells dramatically increased at the start of stationary phase, reaching levels 3 to 4 times that observed in control and nitrogen-limited cultures. These results cannot be explained by changes in the average cell volume. Eventhough dramatic effects on the total toxin concentration were observed in response to nutrient limitation (N or P), the toxin composition (on a percent basis) remained constant. This suggests that the individual toxin composition of a given isolate is a fixed genetic trait and not a transient response to changing environmental factors.     

Uptake,transformation, and elimination kinetics of paralytic shellfish toxins in white seabream (<Emphasis Type="Italic">Diplodus sargus</Emphasis>)

Marine toxins generated by harmful algal blooms can be transferred through the marine food web and ultimately cause massive deaths of piscivorous predators. However, very few studies have explored the processes of accumulation and biotransformation of paralytic shellfish toxins (PSTs) within fishes. White seabream (Diplodus sargus) were orally challenged with contaminated cockles (Cerastoderma edule) containing N-sulfocarbamoyl and decarbamoyl toxins and non-contaminated cockles afterwards. Specific PSTs that occurred in low abundance in cockles (B1 7.6% and dcSTX 1.6% molar fraction) were the only toxins detected in fish viscera possibly resulting from selective elimination and transformation of the various PSTs. Concentration of toxins progressively increased in fish viscera throughout the uptake period. Toxins were then rapidly depurated (B1 0.905 day⁻¹, dcSTX 0.467 day⁻¹) when diet was changed to non-toxic cockles. Results indicate conversion of a precursor toxin into B1 which in turn might be converted into dcSTX at a lower extent. Low accumulation efficiency of 1.7 and 5.0% was calculated to B1 and dcSTX, respectively. This study contributes to a better understanding of dynamics of PSTs in fish and the fate of these compounds in the marine food web.     

Population differences in nerve resistance to paralytic shellfish toxins in softshell clam, <Emphasis Type="Italic">Mya arenaria</Emphasis>, associated with sodium channel mutations

The softshell clam, Mya arenaria, is a commercially important bivalve with wide latitudinal distribution in North America. Populations of clams with a history of repeated exposure to toxic Alexandrium spp. have developed a natural resistance to the paralytic shellfish toxins (PSTs) produced by these algae. An association between PST resistance in individual clams and a single mutation in the saxitoxin (STX) binding region of the α-subunit of the voltage-gated sodium (Na⁺) channel gene was previously identified. Here we establish that more than one mutation associated with nerve resistance to STX occurred at this locus. Both cDNA from mRNA and genomic DNA sequences from individual clams are identical demonstrating that both alleles are expressed simultaneously. In addition, one resistant allele per individual is sufficient to confer neural resistance to STX even though heterozygous individuals show an intermediate level of resistance to STX in in vitro nerve trunk assays.     

Accumulation of paralytic shellfish poisoning toxins in planktonic copepods during a bloom of the toxic dinoflagellate<Emphasis Type="Italic"> Alexandrium tamarense</Emphasis> in Hiroshima Bay,western Japan

Concentrations of paralytic shellfish poisoning (PSP) toxins in toxic dinoflagellate cells and in marine planktonic copepods were monitored during the bloom of Alexandrium tamarense in Hiroshima Bay, western Japan. Concentration of the toxins retained by copepods was a function of the ambient toxin concentration, i.e. the product of A. tamarense cell density and cellular toxicity. The toxin concentration in copepods increased with the increase of toxicants in the seawater then leveled off, but decreased significantly at higher concentrations. In the field, the maximum toxin concentration was 1.2 pmol ind^-1, whereas in the laboratory, the copepod Acartia omorii accumulated a much higher concentration of PSP toxins (24 pmol ind^-1). Feeding avoidance against Alexandrium tamarense and a shift to alternative food sources such as diatoms in the field might keep their toxin levels lower than their potentially maximum level. The copepod toxin levels in the field were not so high as to cause an instantaneous lethal effect on their predator fishes but may reach possibly lethal levels after a few days' continuous feeding. Overall toxin retention by copepods after 12 h feeding and 2 h starvation was only 2.5% of total ingested toxins, which suggested that a significant amount of toxins was released into the seawater. Measurements of toxin reduction and gut evacuation suggested that the toxins were removed through both fecal evacuation and metabolism (e.g. excretion, decomposition and transformation). The results, as a whole, imply that copepods can be a link for PSP toxin flux in both pelagic and benthic food webs and can also be a sink for toxins by metabolizing and removing them from the environment.Communicated by T. Ikeda, Hakodate     

Determination of F2-isoprostanes in cultured human lung epithelial cells after exposure to metal oxide and silica nanoparticles by high-performance liquid chromatography/tandem mass spectrometry

The environmental impact of nanotechnology has caused a great concern. Many in vitro studies showed that many types of nanoparticles were cytotoxic. However, whether these nanoparticles caused cell membrane damage was not well studied. F₂-isoprostanes are specific products of arachidonic acid peroxidation by nonenzymatic reactive oxygen species and are considered as reliable biomarkers of oxidative stress and lipid peroxidation. In this article, we investigated the cytotoxicity of different nanoparticles and the degree of cellular membrane damage by using F₂-isoprostanes as biomarkers after exposure to nanoparticles. The human lung epithelial cell line A549 was exposed to four silica and metal oxide nanoparticles: SiO₂ (15 nm), CeO₂ (20 nm), Fe₂O₃ (30 nm), and ZnO (70 nm). The levels of F₂-isoprostanes were determined by using high-performance liquid chromatography/mass spectrometry. The F₂-isoprostanes’ peak was identified by retention time and molecular ion m/z at 353. Oasis HLB cartridge was used to extract F₂-isoprostanes from cell medium. The results showed that SiO₂, CeO₂, and ZnO nanoparticles increased F₂-isoprostanes levels significantly in A549 cells. Fe₂O₃ nanoparticle also increased F₂-isoprostanes level, but was not significant. This implied that SiO₂, CeO₂, ZnO, and Fe₂O₃ nanoparticles can cause cell membrane damage due to the lipid peroxidation. To the best of our knowledge, this is the first report on the investigation of effects of cellular exposure to metal oxide and silica nanoparticles on the cellular F₂-isoprostanes levels.     

Decomposition of silicate minerals by Bacillus mucilaginosus in liquid culture

The extraction of K⁺ and SiO₂ from silicate minerals by Bacillus mucilaginosus in liquid culture was studied in incubation experiments. B. mucilaginosus was found to dissolve soil minerals and mica and simultaneously release K⁺ and SiO₂ from the crystal lattices. In contrast, the bacterium did not dissolve feldspar. B. mucilaginosus also produced organic acids and polysaccharides during growth. The polysaccharides strongly adsorbed the organic acids and attached to the surface of the mineral, resulting in an area of high concentration of organic acids near the mineral. The polysaccharides also adsorbed SiO₂ and this affected the equilibrium between the mineral and fluid phases and led to the reaction toward SiO₂ and K⁺ solubilization. These two processes led to the decomposition of silicate minerals by the bacterium.     

Photodegradation of the antibiotic spiramycin studied by high-performance liquid chromatography-electrospray ionization-quadrupole time-of-flight mass spectrometry

The degradation of spiramycin induced by ultraviolet (UV) irradiation was studied at pH 3–4, 6–7 and 8–9 using high-performance liquid chromatography-electrospray ionization-quadrupole time-of-flight mass spectrometry. Furthermore, the influence of hydrogen peroxide and oxygen on the photodegradation was investigated. Three degradation products were observed in the presence of oxygen and four in the absence of oxygen. The degradation kinetics were monitored as mass area-time curves, analyzed and described using subsequent follow-up reaction first-order models. The m/z values of the degradation products were determined to be 160, 322 and 366, the latter representing two separate products, the sugar moieties of spiramycin. Remainders of the macrocycle or the intact lactone ring of spiramycin were not observed, since the conjugated π-electron system was assumed photoexcited and thus undergoing photodegradation and fast destruction of the ring. The kinetic of the degradation of spiramycin itself was described using a first-order model. The degradation was accelerated upon addition of hydrogen peroxide. In contrast, systematic acceleration depending on pH could not be found. The absence of oxygen did not lead to any significant acceleration. The findings are interpreted in terms of the effectiveness of advanced oxidation processes and their potential use as a fourth purification step in wastewater treatment plants.     

Field depuration and biotransformation of paralytic shellfish toxins in scallop<Emphasis Type="Italic"> Chlamys nobilis</Emphasis> and green-lipped mussel<Emphasis Type="Italic"> Perna viridis</Emphasis>

Under laboratory conditions, the scallop Chlamys nobilis and the mussel Perna viridis were exposed to N-sulfocarbamoyl toxins (C2 toxin), a paralytic shellfish toxin (PST), by feeding a local toxic strain of the dinoflagellate Alexandrium tamarense (ATDP) that produced C2 toxin exclusively. The bivalves were subsequently depurated in the field, and their depuration kinetics, biotransformation and toxin distribution were quantified. Depuration was characterized by a rapid loss within the first day, followed by a secondary slower loss of toxins. In the fast depuration phase, scallops detoxified PSTs more quickly than the mussels (depuration rate constants for scallops and mussels were 1.16 day^–1 and 0.87 day^–1, respectively). In contrast, the mussels detoxified PSTs more quickly than the scallops in the slow depuration phase, and the calculated depuration rate constants (mean+SE) from day 2 to day 13 were 0.063+0.009 day^–1 and 0.040+0.019 day^–1 for mussels and scallops, respectively. The differences in the appearances of gonyautoxins, GTX2 and GTX3, and their decarbamoyl derivatives, dcGTX2, dcGTX3 and GTX5, which are all derivatives of C2 toxin, indicated active and species-specific biotransformation of the algal toxins in the two bivalves. In both species of bivalves, the non-viscera tissue contained fewer toxins and lower concentrations than the viscera-containing tissue compartment. In scallops, very little toxin was distributed in the adductor muscle. In mussels, most of the PSTs were found in the digestive gland with significant transport of toxins into the digestive gland from other tissues during the course of depuration. The toxin profiles of scallops and mussels differed from each other and from that of the toxic algae fed. A significant fraction of GTX5 was detected in the mussels but not in the scallops. Our study demonstrates a species specificity in the depuration kinetics, biotransformation and tissue distribution of PSTs among different bivalves.Communicated by T. Ikeda, Hakodate     

Primary production and release of assimilated carbon by Chlamydomonas provasolii in culture

Radionuclide labeled sodium bicarbonate was used to measure the rates of primary production and release of metabolites by Chlamydomonas provasolii. The total carbon fixed was 23.18 nM (mg dry wt)^-1 h^-1 and approximately 5.56% of the total was released in organic form into the medium. Thin layer chromatography indicated that the materials released were the amino acids, glycine and leucine. These substances are similar to those identified in certain coral and flatworm symbioses with algae.     

杭州湾南岸滩涂环境状况及与贝类养殖的关系

浙江省慈溪市滩涂环境与贝类养殖的调查结果显示，杭州湾海域水中石油类及无机氮含量介于Ⅱ、Ⅲ类海水水质标准之间，无机磷平均含量超过Ⅳ类海水水质标准。滩涂底质总体上符合海洋沉积物第一类标准，但一些样点多氯联苯（PCBs）超标。分离N2976株细菌，隶属17个属及肠杆菌科的部分属，具有明显的陆源性特点。发现浮游生物71种，主要为硅藻类等广盐性种类，大部分为贝类的主要天然饵料。石油类、Cd、Cr、Hg、Cu及PCBs残留量在一些贝体内不同程度地超过GB18421-2001《海洋生物贝类质量标准》。为有效改良和保育滩涂环境，提高养殖贝类的产量和质量，在揭示滩涂环境与贝类养殖关系的基础上，提出了一些统筹兼顾的措施和建议。     

pH effect on the susceptibility to parasitoid infection in the marine diatom Coscinodiscus spp. (Bacillariophyceae)

The pH on the frustule of individual cells of the marine centric diatoms Coscinodiscus granii and Coscinodiscus wailesii (Bacillariophyceae) was measured with pH microsensors in culture media with increasing pH values of 8.04, 8.14, and 8.22, respectively. In 85–96% of the C. granii cells the pH on the frustule was up to 0.4 units higher than that of the medium, reaching a maximum pH 8.95. Only in 2–3% the surface pH exceeded that of the medium by up to 0.7 pH units. These results strongly suggest that diatoms in batch cultures differ, at least temporarily, in their individual photosynthetic activities. Infection experiments with the parasitoid nanoflagellate Pirsonia diadema (Stramenopile) showed that flagellates failed to infect when the culture pH was 8.8 and above. pH measurements on freshly infected C. granii showed that the prevalence of infection was higher in tendency on diatoms with low surface pH. Application of these results to parasitoid-diatom interactions in natural waters suggests that within phytoplankton populations a strong photosynthetic activity might prevent diatom cells temporarily from infection by pH-sensitive parasitoids.     

酱油曲霉（Aspergillus sojae）利用甘蔗渣产絮凝剂条件筛选及优化

通过Plackett-Burman实验（PB实验）对酱油曲霉（Aspergillus sojae）利用甘蔗渣分泌微生物絮凝剂的8个营养条件和培养条件进行考察,筛选出影响微生物生长、微生物絮凝剂质量和产量的显著性因素,并对显著性因素进行单因子实验,优化酱油曲霉利用甘蔗渣分泌絮凝剂的最佳营养条件和培养条件。实验结果表明：K2HPO4、还原性糖质量浓度、培养时间是影响菌体生长的显著因素,培养基的初始pH是影响絮凝率的显著因素,还原性糖质量浓度、（NH4）2SO4是影响絮凝剂粗产量的显著因素。对显著因素进行单因子实验确定最佳营养条件和培养条件为甘蔗渣的酶解液（还原性糖质量浓度（13.67±0.54）g.L-1）,4 g.L-1的硫酸铵,pH=5,于30℃培养60 h。     

Human transformities in a global hierarchy: Emergy and scale in the production of people and culture

In emergy research, transformities are of fundamental importance. They are factors that are used to convert the inputs to a process into emergy. Once placed in emergy units, the inputs to any process can then be added together or compared. Furthermore, as a product of an emergy analysis, new transformities for outputs can be used in other analyses. By this process the collection of known transformities grows, and subsequent emergy analyses become more accurate. Human labor is often a critical input to an emergy analysis. Transformities for humans have only been roughly estimated based on education level, and should be judged as first approximations. This paper refines the existing values for human services, using similar techniques, but with some different assumptions. The result is a larger range of human transformities, expanded at both lower and upper ends that range from 7.53E4 to 7.53E13. There are many applications of this knowledge, from improving empirical studies to expositions of hierarchy that more reliably “locate” humans, economic production, and information within energy transformation hierarchies.     

Nutrient-limited toxin production and the dynamics of two phytoplankton in culture media: A mathematical model

In this paper we have proposed and analyzed a simple mathematical model consisting of four variables, viz., nutrient concentration, toxin producing phytoplankton (TPP), non-toxic phytoplankton (NTP), and toxin concentration. Limitation in the concentration of the extracellular nutrient has been incorporated as an environmental stress condition for the plankton population, and the liberation of toxic chemicals has been described by a monotonic function of extracellular nutrient. The model is analyzed and simulated to reproduce the experimental findings of Graneli and Johansson [Graneli, E., Johansson, N., 2003. Increase in the production of allelopathic Prymnesium parvum cells grown under N- or P-deficient conditions. Harmful Algae 2, 135–145]. The robustness of the numerical experiments are tested by a formal parameter sensitivity analysis. As the first theoretical model consistent with the experiment of Graneli and Johansson (2003), our results demonstrate that, when nutrient-deficient conditions are favorable for the TPP population to release toxic chemicals, the TPP species control the bloom of other phytoplankton species which are non-toxic. Consistent with the observations made by Graneli and Johansson (2003), our model overcomes the limitation of not incorporating the effect of nutrient-limited toxic production in several other models developed on plankton dynamics.