Hailong Zhou

Oxidative stress, DNA damage and antioxidant enzyme activities in the pacific white shrimp (Litopenaeus vannamei) when exposed to hypoxia and reoxygenation.

To evaluate the genotoxic and physiological effects of acute hypoxia on the pacific white shrimp (L. vannamei), shrimps were treated firstly with three dissolved oxygen levels 6.5 ppm (control), 3.0 ppm and 1.5 ppm for 24 h, respectively, and then reoxygenated (6.5 ppm) for 24 h. The changes of superoxide dismutase (SOD) activity, glutathione peroxidases (GPX) activity, malondialdehyde (MDA) concentration and DNA damage in the tissues of gill, hepatopancreas and hemolymph were examined during the period of hypoxia and reoxygenation. The results indicated SOD activity, GPX activity, MDA concentration and DNA damage all increased basically compared with the control during the period of hypoxia except for MDA concentrations in the gill at 12 h and 24 h hypoxia (3.0 ppm), and these parameters were recovered to some degree during the period of reoxygenation. Moreover, the comet assays in the tissues of gill and hepatopancreas showed an obvious time- and dose-dependent response to hypoxia, which indicated comet assay in the two tissues could be used as sensitive biomarker to detect the occurrence of hypoxia. We conclude that acute hypoxia can induce oxidative stress, DNA damage and lipid peroxidation in the tissues of gill, hepatopancreas and hemolymph of L. vannamei and the DNA damage may come from hypoxia-induced oxidative stress.

Spatial distribution, sources and ecological risk assessment of polycyclic aromatic hydrocarbons in surface seawater from Yangpu Bay, China

The occurrence of 14 polycyclic aromatic hydrocarbons (PAHs) listed by the United States Environmental Protection Agency was investigated in surface seawater of Yangpu Bay, China in November 2013 (winter) and May 2014 (summer). Seventy-two samples were collected from 12 sampling sites. The total concentrations of PAHs in Yangpu Bay showed obvious variations in different seasons, which varied from 582.8 to 2208.3 ng L(-1) in winter and 952.4 to 1201.7 ng L(-1) in summer, respectively. Two-ring PAHs accounted for more than 91.6% of total PAHs in winter, and three-ring PAHs were dominant with 81.6% of total PAHs in summer. Molecular indices analyses indicated that the main source of PAHs in Yangpu Bay could be petrogenic contamination. The ecological risk assessment by Risk quotients (RQNCs and RQMPCs) showed a potential ecological risk of PAHs in Yangpu Bay, indicating a close attention should be paid to pollution of PAHs in the coastal area of Yangpu.

Toxic effects of male Perna viridis gonad exposed to BaP, DDT and their mixture: A metabolomic and proteomic study of the underlying mechanism.

Benzo(a)pyrene and dichlorodiphenyltrichloroethane are typical persistent organic pollutants, and also the widespread environmental estrogens with known toxicity towards green mussels Perna viridis. In this study, the toxicological effects of BaP and DDT and their mixture were assessed in green mussel gonads using proteomic and metabolomic approaches. Metabolomics by NMR spectroscopy revealed that BaP did not show obvious metabolite changes in the gonad of male green mussel. DDT mainly caused some disturbance of osmotic regulation and energy metabolism by changing BCAAs, alanine, threonine, arginine, etc., unknown metabolite (3.53 ppm), glycine, homarine and ATP at different levels. However, the mixture of BaP and DDT mainly caused some disturbance in osmotic regulation and energy metabolism by differentially altering branched chain amino acids, glutamate, alanine, arginine, unknown metabolite (3.53 ppm), glycine, 4-aminobutyrate, dimethylglycine, homarine and ATP. The results suggest that DDT alone may cause most of metabolites changes in the mixture exposed male mussel gonad, and the results also show that the male P. viridis gonad was more sensitive to DDT than BaP exposures. Proteomic study showed that BaP, DDT and their mixture may have different modes of action. Proteomic responses revealed that BaP induced signal transduction, oxidative stress, spermatogenesis, etc. in the male green mussel gonad; whereas DDT exposure altered proteins that were associated with signal transduction, oxidative stress, cytoskeleton and cell structure, cellular organization, energy metabolism, etc. However, the mixture of BaP and DDT affected proteins related to cytoskeleton and cell structure, oxidative stress, cellular organization, etc. This research demonstrated that metabolomic and proteomic approaches could better elucidate the underlying mechanism of environmental pollutants gonad toxicity.

Proteomic and metabolomic analysis on the toxicological effects of Benzo[a]pyrene in pearl oyster Pinctada martensii.

Benzo[a]pyrene (BaP) is one of the typical toxic polycyclic aromatic hydrocarbons (PAHs) that are widely present in marine environment. BaP has diverse toxic effects, including teratogenic, carcinogenic, mutagenic effects and so on, in various organisms. In this work, we focused on the differential proteomic and metabolomic responses in the digestive gland of pearl oyster Pinctada martensii exposed to two doses of BaP (1 and 10μg/L). Metabolic responses revealed that the high dose of BaP (10μg/L) mainly caused disturbances in osmotic regulation and energy metabolism in the digestive gland. Proteomic responses indicated that both doses of BaP induced disturbances in energy metabolism, cytoskeleton, cell injury, oxidative stress and signal transduction based on the differential proteomic biomarkers. Overall, these results demonstrated a number of potential biomarkers that were characterized by an integrated proteomic and metabolomic approach and provided a useful insight into the toxicological effects on pearl oyster P. martensii.

Spatial and temporal distribution and risk assessment of polycyclic aromatic hydrocarbons in surface seawater from the Haikou Bay, China.

Spatial and temporal distributions of 14 polycyclic aromatic hydrocarbons (PAHs) were investigated in surface waters of Haikou Bay, China from October 2013 to September 2014. The total PAHs concentrations ranged from 420.2 to 2539.1 ng L(-1) with the average value of 1016.3±455.8 ng L(-1), which were predominated by low molecular weight PAHs (2- and 3-ring PAHs). Moreover, PAHs displayed spatial and temporal variations in the concentration and composition pattern. Source analysis based on isomer ratios indicated that the PAHs mostly originated from petroleum and combustion processes. An eco-toxicological risk assessment showed that the potential risk of individual PAHs had reached moderate to high levels and the total concentrations of PAHs had also reached a relatively high level compared with previous studies. This study offers important information on the pollution levels of 14 PAHs in the surface waters of Haikou Bay and recommends that prevention and control of PAHs pollution should be implemented in the region.

Transcriptomic responses of Perna viridis embryo to Benzo(a)pyrene exposure elucidated by RNA sequencing.

The green mussel Perna viridis is an ideal biomonitor to evaluate marine environmental pollution. Benzo(a)pyrene (BaP) is a typical polycyclic aromatic hydrocarbon (PAH), which is well known for the mutagenic and carcinogenic characteristics. However, the toxicological effects of BaP on Perna viridis embryo are still unclear. In this study, we investigated the embryo transcriptomic profile of Perna viridis treated with BaP via digital gene expression analysis. A total of 92,362,742 reads were produced from two groups (control and BaP exposure) by whole transcriptome sequencing (RNA-Seq). Gene Ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analysis were used on all genes to determine the biological functions and processes. Genes involved in various molecular pathways of toxicological effects were enriched further. The differential expression genes (DEGs) were related to stress response, infectious disease and innate immunity. Quantitative real-time PCR (qRT-PCR) measured expressional levels of six genes confirmed through the DGE analysis. This study reveals that RNA-seq for transcriptome profiling of P. viridis embryo can better understand the embryo toxic effects of BaP. Furthermore, it also suggests that RNA-seq is a superior tool for generating novel and valuable information for revealing the toxic effects caused by BaP at transcriptional level.

Comparative studies of hemolymph physiology response and HIF-1 expression in different strains of Litopenaeus vannamei under acute hypoxia

Litopenaeus vannamei has a high commercial value and is the primary cultured shellfish species globally. In this study, we have compared the hemolymph physiological responses between two L. vannamei strains under acute hypoxia. The results showed that hemocyanin concentration (HC) of strain A6410 was significantly higher than strain Zhengda; Total hemocyte counts (THC) decreased significantly in both strains under hypoxic stress (p < 0.05). We also investigated the temporal and spatial variations of hypoxia inducible factors 1 (HIF-1) by qRT-PCR. The results showed that hypoxia for 12 h increased the expression levels of HIF-1α in tissues of muscle and gill from the two strains (p < 0.05). In the hepatopancreas, the expression levels of HIF-1 increased significantly in strain Zhengda and decreased significantly in strain A6410 (p < 0.05). No significant changes of HIF-1 expression were detected in the same tissues between the two strains under hypoxia for 6 h (p > 0.05), but in the gills and hepatopancreas under hypoxia for 12 h (p < 0.05). Additionally, the expression level of HIF-1 was higher in the strain Zhengda than A6410 in the same tissue under hypoxia for 12 h. It was indicated that the hypoxic tolerance of Litopenaeus vannamei was closely correlated with the expression level of HIF-1, and the higher expression level of HIF-1 to hypoxia, the lower tolerance to hypoxia in the early stage of hypoxia. These results can help to better understand the molecular mechanisms of hypoxic tolerance and speed up the selective breeding process of hypoxia tolerance in L. vannamei.

Polycyclic aromatic hydrocarbons in surface sediment from Yangpu Bay, China: Distribution, sources and risk assessment

The study investigated the occurrence of polycyclic aromatic hydrocarbons (PAHs) in the surface sediment from eleven sites in Yangpu Bay, China in December 2013 (winter) and July 2014 (summer). The 16 US EPA priority PAHs were found in the range of 1583.2-5701.7 ng/g dry weights with an average of 3134.7 ± 1241.3 ng/g in winter and ranged from 2161.8 to 4527.2 ng/g with an average of 3016.6 ± 748.0 ng/g in summer, respectively. The concentrations of the PAHs tended to be relatively high in comparison with other areas from the literatures. The identification using molecular indices analysis indicated that the PAHs originated mainly from pyrogenic and petrogenic sources in most of the sites. According to principle component analysis-multiple linear regression (PCA/MLR) for their source apportionment, the main sources of PAHs were vehicle emissions, petroleum products and biomass combustion. The risk assessment using international sediments quality guidelines and sediments quality criteria indicated that several PAHs, such as Nap, Flu, Phe, Ace, Acy and BghiP in most of the sites would potentially affect organisms in Yangpu Bay.

Sensitivity of Larvae and Adult and the Immunologic Characteristics of Litopenaeus vannamei under the Acute Hypoxia

Litopenaeus vannamei is one of the most commercially important species of shrimp in the world. In this study, we performed acute hypoxia tests with Litopenaeus vannamei to estimate 12 h median lethal concentration (LC50) values at different life stages. The results indicated that the 12 h LC50 values were significantly different in different life stages of shrimp (p < 0.05). The maximum value of 12 h LC50 was 2.113 mg L-1 for mysis III, and the minimum value was 0.535 mg L-1 for adult shrimp with an average total length of 6 cm. The study also determined the hemocyanin concentration (HC) and the total hemocyte counts (THC) in the conditions of hypoxia and reoxygenation. These results showed that the THC decreased and the HC increased under hypoxia, and the THC increased and the HC decreased in the condition of reoxygenation. These results can provide fundamental information for shrimp farming and seedling and also can guide the breeding selection, as well as being very helpful to better understand the hypoxia stress mechanism of shrimp.

An integrated metabolomic and proteomic study of toxic effects of Benzo[a]pyrene on gills of the pearl oyster Pinctada martensii

Benzo[a]pyrene (BaP) is one of the most important polycyclic aromatic hydrocarbons (PAHs), which are widely present in the marine environment. Because of its teratogenic, mutagenic, and carcinogenic effects on various organisms, the toxicity of BaP is of great concern. In this study, we focused on the toxic effects of BaP (1 µg/L and 10 µg/L) on gills of the pearl oyster Pinctada martensii using combined metabolomic and proteomic approaches. At the metabolome level, the high concentration of BaP mainly caused abnormal energy metabolism, osmotic regulation and immune response marked by significantly altered metabolites in gills. At the proteome level, both concentrations of BaP mainly induced signal transduction, transcription regulation, cell growth, stress response, and energy metabolism. Overall, the research demonstrated that the combination of proteomic and metabolomic approaches could provide a significant way to elucidate toxic effects of BaP on P. martensii.