Jian-Wen Qiu

Acute toxicities of five commonly used antifouling booster biocides to selected subtropical and cosmopolitan marine species

Since 1990s, various booster biocides have been increasingly used as substitutes of organotins. However, knowledge about their toxicities on tropical/sub-tropical marine species is significantly lacking. This study comprehensively investigated the acute toxicities of copper, tributyltin (TBT), and five commonly used booster biocides including Irgarol, diuron, zinc pyrithione (ZnPT), copper pyrithione (CuPT) and chlorothalonil on the growth or survival of 12 marine species in which eight of them are native species of subtropical Hong Kong. We found that Irgarol was more toxic than TBT on the growth of autotrophic species. The toxicity of CuPT was comparable to that of TBT on almost all test species, while it showed higher toxicity than TBT on medaka fish larvae. As the usage of these biocides is expected to further increase worldwide, accurate assessments of their ecological risks are required for better informed decision on their management. This study provided useful datasets for such purposes.

Insights from an Integrated View of the Biology of Apple Snails (Caenogastropoda: Ampullariidae)

ABSTRACT Apple snails (Ampullariidae) are among the largest and most ecologically important freshwater snails. The introduction of multiple species has reinvigorated the field and spurred a burgeoning body of research since the early 1990s, particularly regarding two species introduced to Asian wetlands and elsewhere, where they have become serious agricultural pests. This review places these recent advances in the context of previous work, across diverse fields ranging from phylogenetics and biogeography through ecology and developmental biology, and the more applied areas of environmental health and human disease. The review does not deal with the role of ampullariids as pests, nor their control and management, as this has been substantially reviewed elsewhere. Despite this large and diverse body of research, significant gaps in knowledge of these important snails remain, particularly in a comparative framework. The great majority of the work to date concerns a single species, Pomacea canaliculata, which we see as having the potential to become a model organism in a wide range of fields. However, additional comparative data are essential for understanding this diverse and potentially informative group. With the rapid advances in genomic technologies, many questions, seemingly intractable two decades ago, can be addressed, and ampullariids will provide valuable insights to our understanding across diverse fields in integrative biology.

Environmental factors influencing the distribution of total and fecal coliform bacteria in six water storage reservoirs in the Pearl River Delta Region, China

The Pearl River Delta (PRD) is one of the most developed and densely populated regions in China. Quantifying the amount of pathogens in the source of drinking water is important for improving water quality. We collected water samples from six major water storage reservoirs in the PRD region in both wet and dry seasons in 2006. Results showed that external environmental factors, such as precipitation, location, as well as the internal environmental factors, i.e., physicochemical properties of the water, were closely related with the distribution of coliforms. Seasonally, the coliform bacterial concentrations in wet season were one to two orders of magnitude greater than those in dry season. Spatially, coliform bacterial levels in reservoirs near urban and industrial areas were significantly higher (p < 0.05) than those in remote areas. Correlation analyses showed that the levels of coliforms had close relationships with pH, temperature, suspended solid, organic and inorganic nutrients in water. Principal components analysis further demonstrated that total coliforms in the reservoirs were closely related with water physicochemical properties, while fecal coliforms were more associated with external input brought in by seasonal runoff.

Effects of food availability, larval source and culture method on larval development of Balanus amphitrite amphitrite Darwin: implications for experimental design

Copyright (c) 1997 Elsevier Science B.V. All rights reserved. Effects of food concentration on larval development of Balanus amphitrite amphitrite Darwin (Crustacea: Cirripedia) were studied in three experiments designed to examine plasticity of survival, time of development, and size: I, larvae from a single brood, cultured individually; II, larvae from multiple broods, cultured individually; III, larvae from multiple broods, cultured in groups (batch cultures). Nauplii were fed the diatom Skeletonema costatum (Grev.) Cleve at six concentrations ranging from 0 to 10 6 cells.ml −1 at 24°C and 30‰ salinity. Food concentration had a clear impact on larval survival and development time for B. a. amphitrite and larvae showed the same trends in survival, duration of development, and growth whether they were obtained from single or multiple parents and whether they were cultured individually or in batches. Nauplii of B. a. amphitrite failed to reach the cypris stage at algal concentrations ≤10 3 cells.ml −1 . Development stopped at nauplius II when larvae were reared at 0-10 2 cells.ml −1 and stopped at nauplius III when larvae were reared at 10 3 cells.ml −1 . Survival at 10 5 and 10 6 cells.ml −1 was higher than at 10 4 cells.ml −1 . Larvae reared at the three highest food concentrations developed to the cypris stage. Development time from nauplius II to cyprid was similar (p>0.10) when larvae were maintained at 10 5 or 10 6 cells.ml −1 , but was significantly longer when reared at 10 4 cells.ml −1 . The sizes of B. a. amphitrite larvae at a particular stage were similar regardless of food concentration, developmental rate, larval source (from single vs. multiple broods) and the method of culture. In future studies that involve B. a. amphitrite larvae, larval source and method of culture can be selected solely on the basis of questions to be addressed.

Proteomic basis of stress responses in the gills of the pacific oyster crassostrea gigas

The Pacific oyster Crassostrea gigas is one of the dominant sessile inhabitants of the estuarine intertidal zone, which is a physically harsh environment due to the presence of a number of stressors. Oysters have adapted to highly dynamic and stressful environments, but the molecular mechanisms underlying such stress adaptation are largely unknown. In the present study, we examined the proteomic responses in the gills of C. gigas exposed to three stressors (high temperature, low salinity, and aerial exposure) they often encounter in the field. We quantitatively compared the gill proteome profiles using iTRAQ-coupled 2-D LC-MS/MS. There were 3165 identified proteins among which 2379 proteins could be quantified. Heat shock, hyposalinity, and aerial exposure resulted in 50, 15, and 33 differentially expressed gill proteins, respectively. Venn diagram analysis revealed substantial different responses to the three stressors. Only xanthine dehydrogenase/oxidase showed a similar expression pattern across the three stress treatments, suggesting that reduction of ROS accumulation may be a conserved response to these stressors. Heat shock caused significant overexpression of molecular chaperones and production of S-adenosyl-l-methionine, indicating their crucial protective roles against protein denature. In addition, heat shock also activated immune responses, Ca(2+) binding protein expression. By contrast, hyposalinity and aerial exposure resulted in the up-regulation of 3-demethylubiquinone-9 3-methyltransferase, indicating that increase in ubiquinone synthesis may contribute to withstanding both the osmotic and desiccation stress. Strikingly, the majority of desiccation-responsive proteins, including those involved in metabolism, ion transportation, immune responses, DNA duplication, and protein synthesis, were down-regulated, indicating conservation of energy as an important strategy to cope with desiccation stress. There was a high consistency between the expression levels determined by iTRAQ and Western blotting, highlighting the high reproducibility of our proteomic approach and its great value in revealing molecular mechanisms of stress responses.

Understanding the regulation of estivation in a freshwater snail through iTRAQ-based comparative proteomics.

The apple snail Pomacea canaliculata is a freshwater gastropod with a remarkable ability to withstand seasonal or unpredictable dry conditions by entering estivation. Studies of P. canaliculata using conventional biochemical and the individual gene approaches have revealed the expressional changes of several enzymes and antioxidative genes in response to estivation and arousal. In this study, we applied iTRAQ-coupled two-dimensional LC-MS/MS to identify and quantify the global protein expression during the estivation and arousal of P. canaliculata. A total of 1040 proteins were identified, among which 701 proteins were quantified and compared across four treatments (i.e., control, active snails; short-term estivation, 3 days of exposure to air; prolonged estivation, 30 days of exposure to air; and arousal, 6 h after resubmergence in water) revealing 53 differentially expressed proteins. A comparison of protein expression profiles across treatments indicated that the proteome of this species was very insensitive to initial estivation, with only 9 proteins differentially expressed as compared with the control. Among the 9 proteins, the up-regulations of two immune related proteins indicated the initial immune response to the detection of stress cues. Prolonged estivation resulted in many more differentially expressed proteins (47 compared with short-term estivation treatment), among which 16 were down-regulated and 31 were up-regulated. These differentially expressed proteins have provided the first global picture of a shift in energy usage from glucose to lipid, prevention of protein degradation and elevation of oxidative defense, and production of purine for uric acid production to remove toxic ammonia during prolonged estivation in a freshwater snail. From prolonged estivation to arousal, only 6 proteins changed their expression level, indicating that access to water and food alone is not a necessary condition to reactivate whole-sale protein expression. A comparison with hibernation and diapause revealed many similar molecular mechanisms of hypometabolic regulation across the animal kingdom.

Transcriptomic and iTRAQ proteomic approaches reveal novel short-term hyperosmotic stress responsive proteins in the gill of the Japanese eel (Anguilla japonica).

UNLABELLED Osmoregulation is critical for the survival of fishes that migrate between freshwater (FW) and seawater (SW). The eel, as a catadromous fish, has been studied for decades to reveal the mechanisms of osmoregulation. These studies, however, have been limited by the lack of a genomic database to decipher the mechanism of osmoregulation at a molecular level. In this study, using high-throughput transcriptomic and proteomic technologies, we have provided the first genome-wide study to identify hyperosmotic responsive proteins in the gills of the Japanese eel. Deep sequencing using the 454 platform produced over 660,000 reads with a mean length of 385 bp. For the proteomic study, we collected gill samples from three different treatment groups of fish that had fully adapted to FW/SW or were transferred from FW to SW for 6h. The respective group of gill proteins were extracted and labeled using an isobaric tag for relative and absolute quantitation (iTRAQ) using LTQ-Orbitrap, a high resolution mass spectrometer. Among the 1519 proteins identified from the gill samples, 96 proteins were differentially expressed between FW and SW adapted fish. Nineteen hyperosmotic responsive proteins were detected (10 up-regulated and 9 down-regulated proteins) after 6h post FW to SW transfer. BIOLOGICAL SIGNIFICANCE The study has provided the most comprehensive, targeted investigation of eel gill proteins to date, and shown the powerfulness of combining transcriptomic and proteomic approaches to provide molecular insights of osmoregulation mechanisms in a non-model organism, eel.

Novel Animal Defenses against Predation: A Snail Egg Neurotoxin Combining Lectin and Pore-Forming Chains That Resembles Plant Defense and Bacteria Attack Toxins

Although most eggs are intensely predated, the aerial egg clutches from the aquatic snail Pomacea canaliculata have only one reported predator due to unparalleled biochemical defenses. These include two storage-proteins: ovorubin that provides a conspicuous (presumably warning) coloration and has antinutritive and antidigestive properties, and PcPV2 a neurotoxin with lethal effect on rodents. We sequenced PcPV2 and studied whether it was able to withstand the gastrointestinal environment and reach circulation of a potential predator. Capacity to resist digestion was assayed using small-angle X-ray scattering (SAXS), fluorescence spectroscopy and simulated gastrointestinal proteolysis. PcPV2 oligomer is antinutritive, withstanding proteinase digestion and displaying structural stability between pH 4.0–10.0. cDNA sequencing and protein domain search showed that its two subunits share homology with membrane attack complex/perforin (MACPF)-like toxins and tachylectin-like lectins, a previously unknown structure that resembles plant Type-2 ribosome-inactivating proteins and bacterial botulinum toxins. The protomer has therefore a novel AB toxin combination of a MACPF-like chain linked by disulfide bonds to a lectin-like chain, indicating a delivery system for the former. This was further supported by observing PcPV2 binding to glycocalix of enterocytes in vivo and in culture, and by its hemaggutinating, but not hemolytic activity, which suggested an interaction with surface oligosaccharides. PcPV2 is able to get into predator’s body as evidenced in rats and mice by the presence of circulating antibodies in response to sublethal oral doses. To our knowledge, a lectin-pore-forming toxin has not been reported before, providing the first evidence of a neurotoxic lectin in animals, and a novel function for ancient and widely distributed proteins. The acquisition of this unique neurotoxic/antinutritive/storage protein may confer the eggs a survival advantage, opening new perspectives in the study of the evolution of animal defensive strategies.

Seasonal changes in imposex and tissue burden of butyltin compounds in Thais clavigera populations along the coastal area of Mirs Bay, China.

We assessed the current status of tributyltin (TBT) contamination of Thais clavigera (Gastropoda) along the coastal area of Mirs Bay, China for the first time. The snail samples were collected from 10 different sites in the summer (June) and winter (December) of 2006, respectively. They were analyzed for imposex status, i.e. relative penis size index (RPSI) and vas deferens sequence index (VDSI), followed by quantification of butyltins in their tissues. Most of the collected females suffered from imposex, but the level of butyltin contamination varied with the distance from Yantian Port, which is currently the fourth busiest container port in the world. The tissue concentration of TBT varied with season. For a particular site, the winter samples in general contained much higher concentration of TBT than the summer samples. RPSI, VDSI and organotin concentrations were higher in T. clavigera collected from sites closer to the Port, such as the Seafood Street and Kat O. Both RPSI and VDSI were positively correlated with the tissue burden of tributyltin. VDSI exhibited little seasonal variability, whereas RPSI showed marked seasonal variability, with lower values in the summer samples. The current results will serve as an important reference for long-term monitoring of butyltin contamination in this area.

Development of a Marine Subtidal Epibiotic Community in Hong Kong: Implications for Deployment of Artificial Reefs

A 2-year study was conducted in Hong Kong to examine the effects of substratum, season and length of submersion on the development of a subtidal epibiotic community using four types of settlement panels (concrete, steel, wood and tyre). The season and length of submersion had a strong influence on the total biomass and on community structure while the type of substratum had very little impact on the total biomass or the structure of the epibiotic community. The season of submersion determined the species composition of the newly submerged surfaces. In the spring and summer, tubeworms were the most abundant. In the autumn and winter, barnacles and tunicates dominated. Community succession was not obvious in the first year of submersion as it was intermingled with strong seasonal settlement, growth and death of barnacles and tunicates. In the second year of submersion, green mussels and tunicates settled and grew to occupy most of the panel surfaces, forming an assemblage that was characteristic of climax communities in the local subtidal waters. The results suggest that the type of construction material has limited impact on the development of epibiotic communities on artificial reefs deployed in Hong Kong; the season of submersion may affect community structure in the early successional stage, but not the characteristics of the climax communities. This study indicates that the type of substratum should not be of concern when deploying artificial reefs in the subtidal waters in this region. The design of artificial reefs should focus more on other physical and economical aspects such as durability, flow dynamics, stability, cost, and effects on the ambient environment.