Concurrent exposure of bottlenose dolphins (Tursiops truncatus) to multiple algal toxins in Sarasota Bay, Florida, USA

Sentinel species such as bottlenose dolphins (Tursiops truncatus) can be impacted by large-scale mortality events due to exposure to marine algal toxins. In the Sarasota Bay region (Gulf of Mexico, Florida, USA), the bottlenose dolphin population is frequently exposed to harmful algal blooms (HABs) of Karenia brevis and the neurotoxic brevetoxins (PbTx; BTX) produced by this dinoflagellate. Live dolphins sampled during capture-release health assessments performed in this region tested positive for two HAB toxins; brevetoxin and domoic acid (DA). Over a ten-year study period (2000–2009) we have determined that bottlenose dolphins are exposed to brevetoxin and/or DA on a nearly annual basis (i.e., DA: 2004, 2005, 2006, 2008, 2009; brevetoxin: 2000, 2004, 2005, 2008, 2009) with 36% of all animals testing positive for brevetoxin (n = 118) and 53% positive for DA (n = 83) with several individuals (14%) testing positive for both neurotoxins in at least one tissue/fluid. To date there have been no previously published reports of DA in southwestern Florida marine mammals, however the May 2008 health assessment coincided with a Pseudo-nitzschia pseudodelicatissima bloom that was the likely source of DA observed in seawater and live dolphin samples. Concurrently, both DA and brevetoxin were observed in common prey fish. Although no Pseudo-nitzschia bloom was identified the following year, DA was identified in seawater, fish, sediment, snails, and dolphins. DA concentrations in feces were positively correlated with hematologic parameters including an increase in total white blood cell (p = 0.001) and eosinophil (p<0.001) counts. Our findings demonstrate that dolphins within Sarasota Bay are commonly exposed to two algal toxins, and provide the impetus to further explore the potential long-term impacts on bottlenose dolphin health.

Preliminary studies on the effect of irradiation as a means of preservation of fish and shellfish

Preliminary investigations on the effect of irradiation on commercially important fish and shell fish like silver pomfret, Bombay duck and prawns were conducted. Irradiated samples had an extended storage life compared to their respective controls even though yellowish or brownish discoloration occurred earlier in irradiated fish. Irradiation enhanced the rate of drip formation. Brine treatment prior to irradiation retarded this rate. Pre-blanching was found to further extend the storage life of irradiated fish.

Blood cells as targets of snake toxins

Snake venoms are mixtures of enzymes and peptides which exert toxicological effects by targeting their substrates or receptors upon envenomation. Snake venom proteins widely affect vascular system including circulating blood cells, coagulation factors, an

Studies on aminotransferase enzymes in fish and shellfish

The distribution of aspartate aminotransferase (AAT) and alanine aminotransferase (ALAT) activities in the skeletal muscle of several fish species is reported. AAT activity is found higher than ALAT activity and that the red muscle has higher aminotransferase activity than the white muscle. It is observed that 2-oxoglutaric acid has a wider scope as an amino group acceptor than pyruvic acid in the skeletal muscle of fish. The significance of transamination& in fish is discussed.

Preliminary studies on predicting the setting season of oysters for the benefit of shellfish farmers

Preliminary findings from a spatfall forecasting programme initiated in April 1981 at Himamaylan River, Negros Occidental are presented. Two main activities are involved: 1) monitoring of daily counts of oyster larvae in the plankton; and 2) monitoring of actual setting of oysters on standardized collectors put in the vicinity of oyster farm sites. Findings indicate that when the count of mature larvae exceeds 5 per 100 ml sample and persists for at least 3 days, spat may be expected to occur shortly afterwards.

Effects of toxins induced by phytoplankton bloom on physiological function of indicator animals in Anzali Lagoon

The present overview summarizes data from one year's study during the period of 1379 to 1380 in the regions by "Anzali" lagoon called "Abkenar" and "Hendkhale". Specimens from this lagoon obtained weekly during mordad and shahrivar mounths (July 21 to september 21). The study included 67 types of 5 phytoplanktonal phylum. In "Abkenar" region Cyanobacters with maximum of 97% and minimum of 64.5% of total combination of phytoplanktones made the dominating combination during the period of study , while in "hendkhale" chyrsophyta with maximum of 89% and minimum of 38.7% of total phytoplanktonal was the dominating figure at the same period of time. Researches on ecological parameteres showed that the avarage dissolved oxygen in -Abkenar" and "hendlchale" regions was 10.7 and 8.0 mg/lit respectivly, also total rate of Phosphat in these regions was 0.085 and 0.15 mg/lit respectivly. This study showed that the rates of Nitrat and Amonium in "Abkenar" region was 0.043 and 0.79 mg / lit while for the same substances in "Ilendkhale" measured 0.08 and 0.7 mg I lit respective. Also the avarage rate of chlorophyll a in these two areas measured 58,38 and 40.45 j.un /ht respectivly. Depending on results of correlation cofficient in "Abkenar" region we had Cyanobacters , water and air temperature , Chlorophyll a and total amount of Phosphat as a poitive correlation with transparency while Amonium and Nitrat showed , a negative correlation , EC and finally dissolved oxygen showed a very low rate of correlation coffiocient. To perform this research 5 genus of Cyanobacteres horn "Anzali" lagoon have been isolated and cultured in a laboratorial conditions Later by using Mouse Bioassay method one of these genus identified as a toxic algae. Levels of LD50 with intra peritoneal injection of toxin on mouse in 24 hours was 660 mg/kg and Levels of LC50 by using Bioassay method on Artemia and Daphnia has been shown 618 and 1000 mg kg respectively. Also the physiological effects of were investigated. algae on two types of Cypronides family called Cyprinus carpi° and Hypophrthabnichthys Resultes of blood analyses of Cypronides who were feeded by toxic algae showed a significant decline (P < 0.05) in white and red blood cells and their hematocrites. Levels of LDH , SGOT and SGPT in their blood serum had a significant increase in porportion to control group (P < 0.05) but there was no evidance a differences in Total Protein levels. Pathological studies show damage and destruction of hepato pancreas and kidney of these fishes. Signs and symptoms of intoxication caused by Cyanobacter called Planktohrix agardlltii in mouse and fish show heptotoxic character. This toxin belongs to cyclicpeptides of microcystines group.

In vivo study on the effects of microcystin extracts on the expression profiles of proto-oncogenes (c-fos, c-jun and c-myc) in liver, kidney and testis of male Wistar rats injected i.v. with toxins

Microcystins (MCs) are a potent liver tumor promoter, possessing potent tumor-promoting activity and weak initiating activity. Proto-oncogenes are known to be involved in the tumor-promoting mechanisms of microcystin-LR. However, few data are available on the effects of MCs oil proto-oncogenes in the whole animal. To investigate the effects of MCs on the expression profile of the proto-oncogenes in different organs, male Wistar rats were injected intravenously with microcystin extracts at a dose of 86.7 mu g MC-LR eq/kg bw (MC-LR eq, MC-LR equivalents). mRNA levels of three proto-oncogenes c-fos, c-jun and c-myc in liver, kidney and testis were analyzed using quantitative real-time PCR at several time points post-injection. Significant induction of these genes at transcriptional level was observed in the three organs. In addition, the increase of mRNA expression of all three genes was much higher in liver than in kidney and testis. Meanwhile, the protein levels of c-Fos and c-Jun were investigated by western blotting. Both proteins were induced in the three organs. However, elevations of protein levels were Much lower than those of mRNA levels. These findings suggest that the expression of c-fos, c-jun and c-myc might be one possible mechanism for the tumor-promoting activity and initiating activity of microcystins. (c) 2008 Published by Elsevier Ltd.

Distribution of toxins in various tissues of crucian carp intraperitoneally injected with hepatotoxic microcystins

An acute toxicity experiment was conducted to examine the distribution and depuration of microcystins (MCS) in crucian carp (Carassius aurutus) tissues. Fish were injected intraperitoneally with extracted MCs at a dose of 200 mu g MC-LR (where L = leucine and R = arginine) equivalent/kg body weight. Microcystin concentrations in various tissues and aquaria water were analyzed at 1, 3, 12, 24, and 48 h postinjection using liquid chromatography coupled with mass spectrometry. Microcystins were detected mainly in blood (3.99% of injected dose at 1 h), liver (1.60% at I h), gonad (1.49% at 3 h), and kidney (0.14% at 48 h). Other tissues, such as the heart, gill, gallbladder, intestine, spleen, brain, and muscle, contained less than 0.1% of the injected MCs. The highest concentration of MCs was found in blood (526-3,753 ng/g dry wt), followed by liver (103-1,656 ng/g dry wt) and kidney (279-1,592 ng/g dry wt). No MC-LR was detectable in intestine, spleen, kidney, brain, and muscle, whereas MC-RR was found in all examined fish tissues, which might result from organ specificity of different MCs. Clearance of MC-RR in brain tissue was slow. In kidney, the MC-RR content was negatively correlated with that in blood, suggesting that blood was important in the transportation of MC-RR to kidney for excretion.

Nitrate and phosphate supplementation to increase toxin production by the marine dinoflagellate Alexandrium tamarense

Alexandrium tamarense toxins have great value in biotechnology research as well as important in connection with shellfish poisoning. The influence of nitrate or nitrate and phosphate supplementation on cell biomass and toxin content were investigated in batch cultures. When cultures at low nitrate (88.2 mu M NaNO3) Were supplemented with 793.8 mu M NaNO3 at day 10 the cell density and cellular toxin contents were increased by 6-29% and 20-76%, respectively, compared with controls, and maximal values were 43,600 cells/ml (day 38) and 0.91 pg/cell (day 31). Supplementation with nitrate at day 14 or with nitrate and phosphate at day 10/14 to the cultures did not increase the cell density compared with the non-supplemented middle nitrate or high phosphate (108 mu M NaH2PO4) cultures, respectively, but increased the cellular toxin contents by an average of 52%. The results showed that supplementation with nitrate or with nitrate and phosphate at different growth phases of the cultures increased toxin yield by an average of 46%. Supplementation with nitrate at selected times to maintain continuous low level of nitrate might contribute to the effective increase of toxin yield of A. tamarense. (c) 2005 Elsevier Ltd. All rights reserved.

A screening study on persistent bioaccumulation toxins (PBT) in fuel combustion soot

Persistent bioaccumulative toxins (PBTs) are organic substances that are persistent, bioaccumulative and can cause severe toxic effects (e.g. potential oncogens, mutagenic, endocrine disrupters) to human health or environment which are the ones that need special attention. PBT chemicals could be released to the environment from several types of sources and are ubiquitous in environment. However, fast and efficiency monitoring and assessment methods to investigate PBTs in environment are still lacking. In this study, a cleaning-up procedure of analyzing PBTs in fuels combustion soot was developed and its performance was assessed through comparing the chromatograms of crude extracts with their cleaned extracts after the cleaning-up procedure. The results showed that polycyclic aromatic compounds (PACs) were the main components in fuel combustion soot and the clean-up procedure developed in this paper can be well used as the method of analyzing PBTs in fuels combustion soot.

DSP等赤潮藻毒素对哺乳类细胞的毒性效应及机制研究

本文研究了腹泻性贝毒的主要组分大田软海绵酸以及几株重要赤潮藻的提取物对四株哺乳类细胞的毒性效应，并探讨了应用细胞毒性检测方法作为赤潮藻毒素毒性检测常规方法的可能性。 结果发现：OA和利玛原甲藻提取物显著地抑制四株细胞的增殖并诱导四株细胞发生凋亡；四株细胞对毒素的敏感性存在一定的差异，人肝癌细胞和小鼠神经瘤细胞较敏感，其次分别为人肝细胞和小鼠皮肤细胞。 小鼠神经瘤细胞对OA反应敏感，细胞毒性检测指标及检测方法灵敏、快速，应用小鼠神经瘤细胞Neuro-2a进行的DSP毒素细胞毒性测试方法具有发展为该类毒素毒性监测常规方法的潜能。 米氏凯伦藻内存在抑制细胞增殖的毒性物质，且该物质是一种具有一定极性的脂溶性物质；该物质能够导致细胞肿胀、破裂，并诱导细胞发生脂质过氧化，导致脂质过氧化产物丙二醛(MDA)的积累。 相关亚历山大藻的去藻过滤液内存在抑制细胞增殖和诱导细胞凋亡的毒性物质，该毒性物质的分子量＞5K，这与本实验室以往的研究结果一致。 　　总之，通过我们的研究发现：DSP等赤潮藻毒素或毒性物质对哺乳类细胞存在毒性影响，且不同毒素的危害机制存在差异，赤潮藻毒素或赤潮藻产生的一些毒性物质威胁人类的健康，应引起我们的高度关注。

Growth characters and photosynthetic capacity of Gracilaria lemaneiformis as a biofilter in a shellfish farming area in Sanggou Bay, China

Experiments on growth characters and ecological functions of the macroalgae Gracilaria lemaneiformis, collected from south China, were conducted in polyculture areas of kelp and filter-feeding bivalve in Sanggou Bay in Weihai City, Shandong, in north China from May 2002 to May 2003. The results of 116 days cultivation showed that the average wet weight of alga increased 89 times from 0.1 to 8.9 kg rope(-1), with an average specific growth rate ( based on wet weight) of 3.95% per day. The most favorable water layer for its growth was 1.0 - 1.8 m below the surface in July and August, with an average specific growth rate of 8.2% per day in 30-day experiments. Photosynthetic activity changed seasonally, with an average of 7.3 mg O-2 g dw(-1) h(-1). The maximum rate (14.4 mg O-2 g dw(-1) h(-1)) was recorded in July, or 19.3 mg CO2 g dw(-1) h(-1), while the minimum (0.40 mg CO2 g dw(-1) h(-1)) was in April. This study indicated that the culture of G. lemaneiformis is an effective way to improve water quality where scallops are cultivated intensively.