Responses of growth and hemolymph quality in juvenile Chinese horseshoe crab Tachypleus tridentatus (Xiphosura) to sublethal tributyltin and cadmium.

Responses of growth endpoints and hemolymph constituents in juvenile Chinese horseshoe crab Tachypleus tridentatus under treatments of 0.01 and 0.1 mg/l tributyltin (TBT) and 0.1 and 1 mg/l cadmium (Cd) were examined in a 12-week experiment. A significant decline in final body weight, final prosomal width, percentage of individuals molted and mean molting time was detected under TBT exposures. While morphological abnormalities of the juveniles between TBT treatments and the control were statistically indistinguishable, significantly higher occurrences of carapace erosion and appendage loss were noted under 0.1 and 1 mg/l Cd exposures. Various hemolymph quality indicators, including hemolymph plasma protein level, amebocyte viability and percentage of granular-spherical state of amebocytes of the juveniles exposed to TBT or Cd were significantly lower than the control. Such a decrease in hemolymph quality suggested deleterious effects of metal contaminant-induced stressors on the health status of the juveniles even at low exposure levels (i.e., 0.01 mg/l TBT and 0.1 mg/l Cd). Changes of hemolymph parameters in juvenile horseshoe crabs were more sensitive than growth performance as well as morphological abnormalities in response to metal stressors, and can be used as an indicator to reflect habitat conditions and contaminant levels.

Ecology of artificial reefs in the subtropics.

The application of artificial reefs (ARs) has a long history, and there is a wealth of information related to the design and performance of ARs in coastal and ocean waters worldwide. However, relatively fewer studies in the literature are focused on the response of benthic communities within the reef areas than those on fish attraction and fish production and on the settlement and colonization of epibiota on the AR structures, especially in the subtropics where seasonal differences and environmental conditions can be large. Recent advances in the understanding of the ecology of ARs in the subtropics are highlighted, with a focus on fish attraction versus fish production, development of epibiota on AR systems and responses of in situ benthic communities in the reef areas. Data are also presented on studies of trophic relationships in subtropical AR systems, and further research areas using analyses of biological traits, stable isotope signatures and fatty acid profiles in investigating the ecology of ARs are proposed.

Responses of biological traits of macrobenthic fauna to a eutrophication gradient in a semi-enclosed bay, China.

The impacts of eutrophication on benthic ecological functions are of increasing concern in recent years. In order to assess the response of macrobenthic fauna to increasing eutrophication, two field sampling surveys were conducted during the summer (July-August 2020) and autumn (October-November 2020) from offshore, nearshore to estuarine sediments in Bohai Bay, northern China. Biological trait analysis was employed for the assessment of macrofaunal samples. The results indicated that there was an increase in the proportion of benthic burrowering or tube-dwelling sediment feeders and taxa with higher larval dispersal ability, but a decrease in the proportion of taxa showing high motility in areas with higher nutrient levels. Seasonal differences were also noted in the shift in biological traits, with a significantly lower similarity among the sampling areas in summer and a higher proportion of carnivorous taxa in autumn. The findings suggested that long-term disturbance can lead to the dominance of smaller body-sized benthic species and reduced sediment quality, impeding ecological recovery of benthic organisms under such harsh environment.

Estrogenic potential of benzotriazole on marine medaka (Oryzias melastigma).

This study, for the first time, assessed the reproductive effects of benzotriazoles, widely used industrial chemicals, on marine fish. Marine medakas (Oryzias melastigma) were exposed to 0.01, 0.1, and 1mg/L benzotriazole for periods of four and 35 days. The results that are obtained showed that the expression levels of CYP1A1 were down-regulated in the liver, gills and intestines of both males and females. Vitellogenin (VTG) was highly induced in the liver, gills and intestine of both male and female marine medaka, and CYP19a was up-regulated in the ovaries especially after being exposed for 35 days. Most importantly, the results of the present study suggest that even at environmentally relevant concentrations detected in the aquatic environment, 0.01 mg/L, benzotriazole also caused notable changes in expression levels of VTG, CYP1A1 and CYP19a. More concerns about the toxicity of benzotriazoles on marine animals should be raised.

Seasonal Changes of Growth, Immune Parameters and Liver Function in Wild Chinese Sturgeons Under Indoor Conditions: Implication for Artificial Rearing.

Seasonality has a significant effect on the physiology of fish, especially the effect of water temperature changes. In the present study, the growth, innate immune parameters and liver function indices of two rescued wild adult Chinese sturgeons under captive conditions were monitored for 1 year. The results showed that the total annual weight loss rate of the male was -4.58% and the total weight gain rate of the female was 24.12%, in which the weight of both individuals registered highly significant differences in summer, fall and winter (p < 0.01). The male Chinese sturgeon also exhibited negative specific growth rates (-0.1 to -0.8%) during spring to fall, whereas positive specific growth rates, ranging from 0.03 to 0.11%, were recorded in the female. Seasonality also affected the innate immune parameters of the two Chinese sturgeons, in which leukocytes had been increasing since spring and C-reactive protein (CRP) content was significantly higher (p < 0.05) in summer than fall in both individuals. The CRP level of the male Chinese sturgeon showed a significant increase from fall to winter (p < 0.05), suggesting that it may have contracted infection or inflammation during this study period. With the analysis of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), transaminase (AST:ALT) ratio, alkaline phosphatase, albumin to globulin ratio and triglycerides, it was found that the liver function of the captive Chinese sturgeons was adversely affected along seasonal changes, with the highest degree of liver impairment in winter. In combining observations from growth performance and changes in innate immune and liver function parameters, the present findings deduced that the male Chinese sturgeon under study was more susceptible to seasonal changes than the female. For better indoor culture of adult Chinese sturgeons, monitoring of hematological parameters to detect early signs of inflammation and liver function abnormality should be conducted with routine veterinary care during prolonged captivity.

Temozolomide delivery to tumor cells by a multifunctional nano vehicle based on poly(ß-L-malic acid).

PURPOSE: Temozolomide (TMZ) is a pro-drug releasing a DNA alkylating agent that is the most effective drug to treat glial tumors when combined with radiation. TMZ is toxic, and therapeutic dosages are limited by severe side effects. Targeted delivery is thus needed to improve efficiency and reduce non-tumor tissue toxicity. METHODS: Multifunctional targetable nanoconjugates of TMZ hydrazide were synthesized using poly(ß-L-malic acid) platform, which contained a targeting monoclonal antibody to transferrin receptor (TfR), trileucine (LLL), for pH-dependent endosomal membrane disruption, and PEG for protection. RESULTS: The water-soluble TMZ nanoconjugates had hydrodynamic diameters in the range of 6.5 to 14.8 nm and ζ potentials in the range of -6.3 to -17.7 mV. Fifty percent degradation in human plasma was observed in 40 h at 37°C. TMZ conjugated with polymer had a half-life of 5-7 h, compared with 1.8 h for free TMZ. The strongest reduction of human brain and breast cancer cell viability was obtained by versions of TMZ nanoconjugates containing LLL and anti-TfR antibody. TMZ-resistant cancer cell lines were sensitive to TMZ nanoconjugate treatment. CONCLUSIONS: TMZ-polymer nanoconjugates entered the tumor cells by receptor-mediated endocytosis, effectively reduced cancer cell viability, and can potentially be used for targeted tumor treatment.

Polybrominated diphenylethers (PBDEs) alter larval settlement of marine benthic polychaetes.

Polybrominated diphenylethers (PBDEs) are found ubiquitously in marine environments worldwide. Sediment is the major sink of PBDEs, with the congener BDE 47 being most abundant. In this study, laboratory experiments were carried out to test the hypothesis that contamination of BDE 47 at environmentally realistic sediment concentrations can alter polychaete larval settlement. Using multiple-choice experiment, settlement of three polychaete species (Pseudopolydora vexillosa, Polydora cornuta, and Capitella sp. I) on four types of spiked sediment was studied and compared: (i) low BDE 47 concentration (0.5 ng g(-1) dry weight); (ii) high BDE 47 concentration (3.0 ng g(-1) dry weight), (iii) hexane (solvent control), and (iv) natural sediment (control). Our results showed that settlement of P. vexillosa and Capitella sp. I larvae was significantly promoted, while settlement of P. cornuta reduced, at high BDE 47 concentration in sediment compared with the respective controls under both short- (24 h) and long-term (4 week) exposures. After 4 weeks, body burden of BDE 47 in all polychaete species was directly related to the spike concentration, and body length of settled juveniles of P. vexillosa and Capitella sp. I at the high-concentration treatment was significantly longer compared with that of other treatments and controls. For the first time, we demonstrated that environmentally realistic concentrations of BDE 47 in sediment can affect polychaete settlement in species-specific and dose-dependent manners. Given the global contamination of PBDE in marine sediment, BDE 47 may potentially alter the settlement pattern of marine polychaetes and hence their benthic composition over large areas.

Changes of substrate microbial biomass and community composition in a constructed mangrove wetland for municipal wastewater treatment during 10-years operation.

Constructed wetlands (CWs) have been used for wastewater treatment for decades, but research on microorganisms involved, especially long-term changes, is still limited. In this study, we evaluated changes in the substrate microbial community in a pilot-scale horizontal subsurface-flow constructed mangrove wetland during 10-years operation. In the 3rd year of operation, microbial biomass carbon and phospholipid fatty acids (PLFAs) reached peak values in two vegetated belts planted with Aegiceras corniculatum (Ac) and Kandelia obovata (Ko), respectively, then stabilized or declined in the 9th and 10th years of operation. PLFA profiles reflecting microbial community compositions varied significantly in the Ac belt during the operation period. Principal component (PCA) and redundancy analyses (RDA) revealed that microbial community compositions were significantly correlated with organic matter content, especially in the 9th and 10th years of operation, implying that the substrate microbial community in constructed mangrove wetland is sensitive to substrate characteristics and can be used as an indicator for long-term performance of CWs.

Toxicity of bisphenol A and its bioaccumulation and removal by a marine microalga Stephanodiscus hantzschii.

The toxicity of bisphenol A (BPA) to Stephanodiscus hantzschii, a diatom isolated from tidal water of Futian Mangrove Nature Reserve, China, and the bioaccumulation and removal capability of the marine microalga to BPA were investigated in the present study. Toxicity experiments showed that the 96-h EC50 of BPA was 8.65+/-0.26 mg/L, and the cell number and chlorophyll a content of S. hantzschii decreased significantly with increases in BPA at concentrations higher than 3.00 mg/L. S. hantzschii had high removal capability at low BPA concentrations as BPA was bioaccumulated and biodegraded by cells. After 16-day treatment, 88%, 99%, 92%, 61%, 48%, 28% and 26% of BPA were removed by the diatom in the media supplemented with 0.01, 0.10, 1.00, 3.00, 5.00, 7.00 and 9.00 mg/L BPA, respectively. The present study demonstrated that S. hantzschii was a tolerant isolate that could be used to remove BPA from contaminated waters.

Deriving site-specific sediment quality guidelines for Hong Kong marine environments using field-based species sensitivity distributions.

Field data of benthic communities and contaminant loadings in marine sediments measured in parallel can be used to derive sediment quality guidelines (SQGs) using a field-based species sensitivity distribution (f-SSD) approach. Recently, SQGs have been successfully derived from f-SSDs for the Norwegian continental shelf with an extensive survey (>1 million km(2)) and a large data set (1,902 sampling stations with 1,944 species). The present study examined the practicality of this approach in deriving SQGs for a much smaller geographical area, namely, the marine environment of Hong Kong (sea area: 1,651 km(2)), making use of databases of the government of Hong Kong special administrative region. As the construction of f-SSDs requires the use of a collection of responses from individual species to a chemical gradient in sediment, data screening criteria on the minimum abundance of the species were evaluated and optimized to ensure sufficient statistical power for estimating these responses. Sediment quality guidelines were derived for nine trace metals, total polycyclic aromatic hydrocarbons, and total polychlorinated biphenyls and compared with current SQGs in developed countries. The community-adjusted hazardous concentrations of 5% and 10% of the f-SSDs were adopted to represent the threshold effects level (TEL) and predicted effects level (PEL), respectively. The TELs derived from this f-SSD approach compares favorably with current SQGs, while the derived PELs were generally lower than the current SQGs, indicating that they are more protective. The f-SSDs can be directly utilized for probabilistic risk assessment, while the field-based SQGs can be used as site-specific guidelines or integrated into current SQGs. Our results suggest that the f-SSD approach can also be applicable to small areas such as Hong Kong.

Amelioration of marine farming impact on the benthic environment using artificial reefs as biofilters.

An in situ monitoring of the sediment characteristics and macrobenthic communities was undertaken at a marine fish culture site in subtropical waters of Hong Kong before and after the deployment of biofilters which were made of cement concrete artificial reef (AR) structures. According to the distance to the boundary of the fish cages, 6 points were selected as sampling stations: 2 at the fish cages, 2 near the boundary of the fish culture area, and 2 reference sites further away from the culture area. Bimonthly sediment samples were collected for analysis of silt-clay fraction (SCF), moisture content (MC), total organic carbon (TOC), total Kjeldahl nitrogen (TKN) and total phosphorus (TP). The macrobenthos (>0.5mm) present in the sediment were sorted, identified and enumerated. TOC, TKN and TP levels at the fish cage stations were consistently higher than those at the reference stations over the 1-year pre-AR and 2-year post-AR deployment monitoring period. The diversity of macrofauna was significantly reduced at the fish cage stations relative to the reference sites. The intermediary stations near the fish culture area showed a transitional state of disturbance. Over the 2-year post-AR deployment period, TOC, TKN and TP showed a decreasing trend at the fish cage and intermediary stations. More diverse macrofaunal communities were recorded at the fish cage stations, with species diversity H'increasing from 0-1 at the beginning of the AR deployment to H'>2 at the end of the study. The present results demonstrated that artificial reefs can improve the benthic abiotic environment and biotic conditions beneath fish rafts which are deteriorated due to farming activities.

Field observations on correlation of fatty acid profiles between suspended particulate matter and green-lipped mussels in subtropical waters of Hong Kong.

By analysis of the fatty acid profiles in mussel tissues and suspended particulate matter (SPM) in water, the present study showed a significant relationship of the trophic linkage between mussels and the SPM. At seven locations from inner to outer areas along the eutrophic Tolo Harbour and Tolo Channel, Hong Kong, the composition (as percentage of total fatty acids) of both monoenoic and polyunsaturated fatty acids in the tissues of green-lipped mussels Perna viridis and SPM in water had significant correlation (p<0.01). In particular, the composition of docosahexaenoic acid (DHA, 22:6n3) (as percentage of total fatty acids) in mussel tissues was statistically correlated with that in SPM (p<0.01), implying that mussels incorporate DHA, the biomarker of dinoflagellates. Principal component analysis further demonstrated that the fatty acid profiles of SPM were different among locations in the harbour, mid and outer channel of the study area, so were the mussel tissue fatty acid profiles. Cluster analysis of phytoplankton data also revealed the dominance of diatoms in the inner harbour and channel areas, whereas dinoflagellates were abundant in the outer channel waters. The possible implications of using benthic suspension feeders such as green-lipped mussels P. viridis for controlling phytoplankton abundance in coastal waters are discussed.

Use of ciliates (Protozoa: Ciliophora) as bioindicator to assess sediment quality of two constructed mangrove sewage treatment belts in Southern China.

To complement physical and chemical data, information of biological communities is important to assess the qualities of mangrove sediments receiving wastewater. Ciliate communities have cosmopolitan distribution, short life cycle and high sensitivity to pollutants, which make them useful as biological indicators of the sediment environment. In most literature, ciliates are widely used as bioindicators for the state of water quality. In this study, the physico-chemical parameters and ciliate community structure of surface sediment collected at different sampling points from two constructed mangrove (Aegiceras and Sonneratia) belts for treatment of municipal sewage in southern China were investigated. Results showed that most (> 80%) of the 216 species ciliates identified at the two constructed mangrove belts were either omnivorous or bacterivorous. Sediment redox potential (Eh) was considered an important factor to govern the distribution of ciliate species within the mangrove sediment. The saprobic system originally derived from freshwater ecosystem was used to evaluate the saprobic degrees of these constructed mangrove belts. Saprobic index (SI) values declined from the sewage inlet to the outlet points of the constructed belts, suggesting better sediment quality at the outlet point caused by treatment processes within the mangrove belt system. Sediment quality of the sewage outlet area of the constructed Aegiceras belt was determined as class II-III (SI = 2.48), while that of the Sonneratia belt was as class III (SI = 2.71) according to the saprobic classification, indicating that a better sewage treatment efficiency was apparent in the Aegiceras than Sonneratia belt. The present data suggested that ciliates could serve as a good bioindicator in assessing organically polluted sediment qualities.

The spatial and temporal distribution of heavy metals in sediments of Victoria Harbour, Hong Kong.

Victoria Harbour has received substantial loadings of pollutants from industrial and municipal wastewater discharged since the 1950s. Inputs of contaminants have declined dramatically during the last two decades as a result of better controls at the source and improved wastewater treatment facilities. To assess the spatial and temporal changes of metal contaminants in sediments in Victoria Harbour, core and grab sediments were collected. The central harbour areas were generally contaminated with heavy metals. The spatial distribution of trace metals can probably be attributed to the proximity of major urban and industrial discharge points, and to the effect of tidal flushing in the harbour. In the sediment cores, the highest concentrations of trace metals were observed to have accumulated during the 1950s-1980s, corresponding with the period of rapid urban and industrial development in Hong Kong. From the late 1980s, there has been a major decline in the concentrations of trace metals, due to a reduction in industrial activities and to the enactment of wastewater pollution controls in the territory. The Pb isotopic compositions of the sediments revealed the anthropogenic inputs of Pb to the harbour. The (206)Pb/(207)Pb ratios varied from 1.154 to 1.190, which were lower than those of background geological materials in Hong Kong ((206)Pb/(207)Pb: 1.201-1.279). The data also indicated that the Pb in the harbour sediments most likely originated from mixed sources, including the leaded gasoline used in the past and other anthropogenic sources.

Trophic and growth baseline of dominant subtidal gastropods in contrasting subtropical marine environments.

Using 13C/12C, 15N/14N and 18O/16O isotopes, the trophic relationship and growth estimation were analyzed in gastropods Nassarius siquijorensis, Murex trapa and Turritella bacillum and their potential food sources and predators in summer and winter from estuarine and oceanic environments in subtropical Hong Kong. Results of δ13C and δ15N values and isotopic mixing model revealed N. siquijorensis and M. trapa were one trophic level higher than T. bacillum, in which its main food source was particulate organic matter (POM) whereas N. siquijorensis largely consumed POM and polychaetes and M. trapa also preyed on other gastropods. Crabs were the major predator of gastropods. Organisms collected from oceanic waters were more 13C enriched than from estuarine waters, reflecting different carbon food sources from marine or terrestrial origin. The δ18O profile from shell carbonate suggested these gastropods were one to two years old. T. bacillum exhibited faster summer growth than the other two species.

Impact of habitat management on waterbirds in a degraded coastal wetland.

The loss of coastal wetlands in Hong Kong Mai Po Nature Reserve adversely affected wetland-depended species. To mitigate this impact, gei wai ponds were reconstructed according to a set of biodiversity management zones (BMZs). This study, based on Canonical Correspondence Analysis (CCA), investigated if waterbird distribution was related to BMZ characteristics. Based on habitat characteristics, ponds in the same BMZ generally clumped in the same quadrant or within a short distance on CCA scatter plots, indicating that a BMZ zone produced common habitat traits. Ponds in a close distance on the plot had similar bird abundance or community structure. Significant correlations were noted between the abundance of cormorants and tall tree, and between waders and bare ground areas within study ponds. This study indicated that the control of key habitat factors was important for the success of reconstruction of gei wais and management of waterbirds in Mai Po.

Does energetic cost for leaf construction in Sonneratia change after introduce to another mangrove wetland and differ from native mangrove plants in South China?

Exotic species invasions are serious ecological problems. Leaf construction cost (CC) and growth traits of two Sonneratia (Sonneratia caseolaris and S. apetala) and four native species (Bruguiera gymnorrhiza, Kandelia obovata, Aegiceras corniculatum and Avicennia marina) in Hainan and Shenzhen mangrove wetlands were compared to evaluate invasive potentials of Sonneratia after introduced to Shenzhen, their new habitat. There were no significant differences in CC and growth traits between two wetlands, suggesting Sonneratia did not lose any advantage in the new habitat and were competitive in both wetlands. CC per unit mass (CCM), CC per unit area (CCA) and caloric values of Sonneratia were significantly lower than those of native mangrove species while specific leaf area (SLA) was just the opposite. CCM of S. caseolaris and S. apetala were 6.1% and 11.9% lower than those of natives, respectively. These findings indicated the invasive potential of Sonneratia in Shenzhen after their introduction.

Performance and bacterial community structure of a 10-years old constructed mangrove wetland.

Constructed mangrove wetland has been used for wastewater treatment but its long-term performance has not been reported. One-year monitoring of a 10-years old horizontal subsurface-flow constructed mangrove wetland consisting of three belts, two with mangrove plants and one without, revealed that the system maintained high and stable removal percentages of organic matter and nutrients, and planted belts performed better than unplanted control. Substrates in belts planted with Aegiceras corniculatum or Kandelia obovata had higher abundance of ammonifiers, nitrifiers and denitrifiers but lower total heterotrophic bacteria than unplanted substrate. Denaturing gradient gel electrophoresis showed that microbial diversity in planted substrate was significantly lower than that in unplanted one. The bacteria in substrates, irrespective to belts, were phylogenetically related to Proteobacteria (most dominant), Acidobacteria, Firmicutes, Nitrospirae, Gemmatimonadetes, Chloroflexi and Cyanobacteria. The steady performance of this 10-year old constructed mangrove wetland was affected by the abundance and diversity of bacterial community in substrate.

Effects of nitrogen and sulphide on macroinfaunal community: a microcosm study.

A microcosm experiment was conducted to test the short-term effects of nitrogen (as TKN, total Kjeldahl nitrogen) and sulphide (as AVS, acid volatile sulphide) on macrobenthic infauna over a period of 8 weeks. The experiment consisted of four treatments using sediment cores (D: 8 cm, H: 20 cm) with addition of: I, nitrogen (N) to an in situ mean level of 0.48 mg Ng(-1) dw; II, sulphidic solution (S) to an in situ mean level of 318.8 microM S g(-1) dw; III, nitrogen and sulphidic solution (NS) to in situ mean levels of 0.45 mg Ng(-1) dw and 329.0 microM S g(-1) dw, respectively, and IV, control with no addition of N and S. Sediment cores were retrieved for analysis of infaunal composition after weeks 2, 4 and 8. A total of 646 specimens of macrobenthic infauna belonging to 27 species were recorded from the cores, in which polychaetes were the most abundant with respect to species and individual numbers. Mean species number and diversity H' of the control and N treatment was statistically higher than S and NS treatments, mean individual number of the S treatment was larger than the NS treatment, and mean evenness J of the S treatment was lower than the N and NS treatments as well as the control. Individual numbers also showed a significant increase from weeks 2 to 8, whereas evenness J decreased in weeks 4 and 8. Multivariate analyses of the faunal data suggested that benthic composition of the N treatment and control did not differ during the experimental period, but changes in benthic structure in S and NS treatments were evident. The present findings demonstrated the dose-response relationship of benthic species changes under controlled addition of N and S. The response to N and S additions in the sediment microcosms was in agreement with the general effects of organic enrichment on macrobenthic communities along a spatial gradient of organic pollution as described by Pearson and Rosenberg [Pearson, T.H., Rosenberg, R., 1978. Macrobenthic succession in relation to organic enrichment and pollution of the marine environment. Oceanography and Marine Biology Annual Review 16, 229-311]. However, in the present experiment, community changes in the treatments were observable in a short, temporal scale.

Sediment characteristics and benthic ecological status in contrasting marine environments of subtropical Hong Kong.

Sediment characteristics and benthic communities on a finer sampling scale in four contrasting environments in subtropical Hong Kong were analyzed in summer and winter 2012. In two harbour habitats which suffered from historic sewage pollution or hypoxic events, organic carbon, nutrient and trace metal content in the sediment were significantly higher than that in an offshore area and a marine reserve. The relatively low organic and nutrient content in the offshore habitat could be resulted from enhanced resuspension of such materials from the seabed owing to intense water mixing and disturbance caused by bottom trawling. The biotic indices AMBI and M-AMBI were shown to be useful in assessing the benthic ecological status of these habitats. Such indices can also be more sensitive than sediment physico-chemical parameters in differentiating the response of macrofauna to seasonal changes in the benthic environment.