Cytotoxicity induced by abamectin in hepatopancreas cells of Chinese mitten crab, Eriocheir sinensis: An in vitro assay.

Toxic effects of abamectin on non-target aquatic organisms have been well documented due to its extensive use in both agricultural and aquacultural areas. However, knowledge of the abamectin induced cytotoxicity in crustacean hepatopancreas is still incomplete. In this study, we investigated the cytotoxic effects of abamectin on hepatopancreas cells of Chinese mitten crab, Eriocheir sinensis by an in vitro assay. The results showed that abamectin inhibited cell viability with elevated reactive oxygen species (ROS) and malondialdehyde (MDA) levels in a dose-dependent manner. Increased olive tail moment (OTM) values and 8-hydroxy-2'-deoxyguanosine (8-OHdG) contents indicate the DNA damage under abamectin exposure. The up-regulation of the typical apoptosis-related protein BCL2-associated X protein (Bax) and the down-regulation of B cell leukemia/lymphoma 2 (Bcl-2) demonstrate apoptosis in hepatopancreas cells. Meanwhile, the activities of both caspase-3 and caspase-9 were increased, indicating caspase-mediated apoptosis. In addition, qRT-PCR results showed the up-regulation of antioxidant genes superoxide dismutase (SOD) and catalase (CAT). The mRNA expression of Cap 'n' Collar isoform-C (CncC) and c-Jun NH2-terminal kinases (JNK) was also significantly increased, implying the involvement of the Nrf2/MAPK pathway in the antioxidative response. The alteration of innate immune-associated genes Toll-like receptor (TLR) and myeloid differentiation primary response gene 88 (Myd88) also indicates the influence of abamectin on immune status. In summary, the present study reveals the cytotoxicity of abamectin on hepatopancreas cells of E. sinensis and this in vitro cell culture model could be used for further assessment of pesticide toxicity.

Effects of chlorantraniliprole-based pesticide on transcriptional response and gut microbiota of the crucian carp, Carassius carassius.

Chlorantraniliprole (CAP) is a presentative diamide pesticide utilized in agricultural area and as well as rice-fish co-culture system for pest control. However, the understanding of toxic effects of CAP on fish species is still incomplete. In the present study, we performed an integrated study of the acute toxicity and bioaccumulation of CAP on the crucian carp, Carassius carassius, a fish species widely distributed in freshwater area in China and commonly farmed in the rice-fish co-culture systems. Besides, biochemical changes, transcriptional responses and gut microbiota of fish were investigated upon sub-chronic CAP exposure. The results showed that CAP is low toxic to crucian carp with a 96 h LC50 of 74.824 mg/L, but has considerable accumulation in the fish muscles when exposed to 3 mg/L of CAP for 14 d and still detectable after 18 d recovery in fresh water. For sub-chronic test, fish were exposed to CAP at 0, 0.3, 3 and 30 mg/L respectively for 14 d. CAP induced oxidative stress and detoxification inhibition in the liver of fish by decreasing antioxidative and detoxicated enzymes activities and downregulating relevant genes expression. In addition, disrupted gut flora composition was found in all experimental groups by the 16 S rRNA sequencing data, indicating the gut microbiota dysbiosis in crucian carp and potential adverse host effect. All the results suggest that CAP at sublethal concentrations has prominent toxic effect on crucian carp and more attentions should be paid especially using directly in an integrated aquaculture system.

Predicting the impact of the COVID-19 pandemic on globalization.

The COVID-19 pandemic has significantly influenced the global economy, international travel, global supply chains, and how people interact, and subsequently affect globalization in coming years. In order to understand the impact of COVID-19 on globalization and provide potential guidance to policymakers, the present study predicted the globalization level of the world average and 14 specific countries in scenarios with and without COVID-19 based on a new Composite Indicator method which contains 15 indicators. Our results revealed that the world average globalization level is expected to decrease from 2017 to 2025 under the scenario without COVID-19 by -5.99%, while the decrease of globalization under the COVID-19 scenario is predicted to reach -4.76% in 2025. This finding implies that the impact of COVID-19 on globalization will not be as severe as expected in 2025. Nevertheless, the downward trend of globalization without COVID-19 is due to the decline of the Environmental indicators, whereas the decline under the COVID-19 scenario is attributed to Economic aspects (almost -50%). The impact of COVID-19 on globalization varies across individual countries. Among the countries investigated, COVID-19 had a positive impact on the globalization of Japan, Australia, the United States, the Russian Federation, Brazil, India and Togo. In contrast, the globalization in the United Kingdom, Switzerland, Qatar, Egypt, China and Gabon are expected to decrease. The variation of impact induced by COVID-19 on those countries is attributed to the weighting of economic, environmental and political aspects of globalization is different across these countries. Our results can help governments take suitable measures to balance economic, environmental and political policies, which may better support their decision-making.

Search for Dark Matter Annihilation Signals in the H.E.S.S. Inner Galaxy Survey.

The central region of the Milky Way is one of the foremost locations to look for dark matter (DM) signatures. We report the first results on a search for DM particle annihilation signals using new observations from an unprecedented γ-ray survey of the Galactic Center (GC) region, i.e., the Inner Galaxy Survey, at very high energies (≳100 GeV) performed with the H.E.S.S. array of five ground-based Cherenkov telescopes. No significant γ-ray excess is found in the search region of the 2014-2020 dataset and a profile likelihood ratio analysis is carried out to set exclusion limits on the annihilation cross section ⟨σv⟩. Assuming Einasto and Navarro-Frenk-White (NFW) DM density profiles at the GC, these constraints are the strongest obtained so far in the TeV DM mass range. For the Einasto profile, the constraints reach ⟨σv⟩ values of 3.7×10^{-26} cm^{3} s^{-1} for 1.5 TeV DM mass in the W^{+}W^{-} annihilation channel, and 1.2×10^{-26} cm^{3} s^{-1} for 0.7 TeV DM mass in the τ^{+}τ^{-} annihilation channel. With the H.E.S.S. Inner Galaxy Survey, ground-based γ-ray observations thus probe ⟨σv⟩ values expected from thermal-relic annihilating TeV DM particles.

DNA damage, immunotoxicity, and neurotoxicity induced by deltamethrin on the freshwater crayfish, Procambarus clarkii.

Pyrethroid pesticides are applied to both agricultural and aquacultural industries for pest control. However, information of their impact on the commercial important freshwater crayfish, Procambarus clarkii is scarce. Therefore, the present study aimed to characterize to effects of a commonly used pyrethroid pesticide, deltamethrin on DNA damage, immune response, and neurotoxicity in P. clarkii. Animals were exposed to 7, 14, and 28 ng/L of deltamethrin, which correspond to 1/8, 1/4, and 1/2 of the LC50 (96 hours) of this pyrethroid to P. clarkii. Significant increase of olive tail moment (OTM) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) was found after deltamethrin exposure in a dose-dependent way. Total hemocyte counts (THC) and activities of immune-related enzymes including acid phosphatase (ACP), lysozyme (LZM), and phenoloxidase (PO) were all decreased and significantly lower than control at concentration of 28 ng/L after 96 hours exposure. Acetylcholinesterase (AChE) activity, an indicator of neurotoxic effect was investigated and it was decreased significantly in muscles at 14 and 28 ng/L after 24 hours exposure. The level of intracellular reactive oxygen species (ROS) in hemocytes was also measured and the significant increase of ROS was found at 14 and 28 ng/L concentrations. The results revealed that deltamethrin induced DNA damage, immunotoxicity, and neurotoxicity in P. clarkii by excessive generation of ROS. Because of the dose-dependent responses of all parameters under exposure of deltamethrin at environmentally realistic concentrations, these parameters could be used as sensitive biomarkers for risk assessment of deltamethrin in aquaculture area.

Oxidative stress, immunological response, and heat shock proteins induction in the Chinese Mitten Crab, Eriocheir sinensis following avermectin exposure.

In this study, the Chinese mitten crabs, Eriocheir sinensis were exposed to avermectin at 0.03, 0.06, 0.12, 0.24, and 0.48 mg/L respectively for 96 hours. The results showed that levels of superoxide dismutase, catalase, and glutathione peroxidase in hepatopancreas were slightly induced at concentration of 0.03 and 0.06 mg/L, but significantly (P < .05) decreased at higher concentrations, meanwhile similar trend of the activities of acid phosphatase, alkaline phosphatase and lysozyme were observed. Significant induction of HSP70 and HSP90 mRNA expression was detected at 24 hours whereas no significant change was found in HSP60. In addition, levels of reactive oxygen species in hepatocytes increased in dose- and time- dependent manners, and cell viabilities of hepatocytes and haemocytes decreased. These results indicated that sublethal concentration exposure of avermectin had a prominent oxidative stress effect on E. sinensis based on the antioxidative and immunological activity inhibition, and HSP60, 70, and 90 perform a protective response during the exposure.

GPS Trajectory Completion Using End-to-End Bidirectional Convolutional Recurrent Encoder-Decoder Architecture with Attention Mechanism.

GPS datasets in the big data regime provide rich contextual information that enable efficient implementation of advanced features such as navigation, tracking, and security in urban computing systems. Understanding the hidden patterns in large amount of GPS data is critically important in ubiquitous computing. The quality of GPS data is the fundamental key problem to produce high quality results. In real world applications, certain GPS trajectories are sparse and incomplete; this increases the complexity of inference algorithms. Few of existing studies have tried to address this problem using complicated algorithms that are based on conventional heuristics; this requires extensive domain knowledge of underlying applications. Our contribution in this paper are two-fold. First, we proposed deep learning based bidirectional convolutional recurrent encoder-decoder architecture to generate the missing points of GPS trajectories over occupancy grid-map. Second, we interfaced attention mechanism between enconder and decoder, that further enhance the performance of our model. We have performed the experiments on widely used Microsoft geolife trajectory dataset, and perform the experiments over multiple level of grid resolutions and multiple lengths of missing GPS segments. Our proposed model achieved better results in terms of average displacement error as compared to the state-of-the-art benchmark methods.

Obtaining P3P privacy policies for composite services.

With the development of web services technology, web services have changed from single to composite services. Privacy protection in composite services is becoming an important issue. P3P (platform for privacy preferences) is a privacy policy language which was designed for single web services. It enables service providers to express how they will deal with the privacy information of service consumers. In order to solve the problem that P3P cannot be applied to composite services directly, we propose a method to obtain P3P privacy policies for composite services. In this method, we present the definitions of Purpose, Recipient, and Retention elements as well as Optional and Required attributes for P3P policies of composite services. We also provide an instantiation to illustrate the feasibility of the method.

Abamectin-induced behavioral alterations link to energy metabolism disorder and ferroptosis via oxidative stress in Chinese mitten crab, Eriocheir sinensis.

The increasing application of abamectin (ABM) in agriculture has raised concerns regarding its environmental safety and potential adverse effects on aquatic environment safety. In the present study, the toxic effects of ABM exposure on the adult Chinese mitten crab, Eriocheir sinensis were investigated, with a focus on locomotion impairment, behavioral changes, oxidative stress, energy metabolism disruption, and ferroptosis. Crabs were exposed to sublethal concentrations of ABM at 2, 20 and 200 µg/L. After 21 d chronic exposure to 200 µg/L, residual ABM in hepatopancreas and muscles were detected as 12.24 ± 6.67 and 8.75 ± 5.42 µg/Kg, respectively. By using acute exposure experiments (96 h), we observed significant locomotion and behavioral alterations, alongside biochemical evidences of oxidative stress and energy metabolism impairment. The presence of ferroptosis, a form of cell death driven by iron-dependent lipid peroxidation, was notably identified in the hepatopancreas. Functional tests with N-acetylcysteine (NAC) supplementation showed restored behavioral responses and decrease of ferroptosis levels. It suggests that mitigating oxidative stress could counteract ABM-induced toxicity. Our findings highlight the critical roles of oxidative stress and ferroptosis in mediating the toxic effects of ABM on E. sinensis, underscoring the need for strategies to mitigate environmental exposure to pesticides.

Functional prediction of AMP deaminase 1 in Jingyuan chicken and evaluation of the biological activities of its expression vectors.

This study investigated the function of AMP deaminase 1 (AMPD1) in Jingyuan chicken and the biological activity of its expression vector. AMPD1 was cloned and sequenced from chicken breast muscle tissue by RT-PCR and further analyzed using Cluster, DNASTAR, and online bioinformatics software, as well as vector construction, qPCR, Western blotting, enzymatic digestion, and sequencing. The coding sequence was 2162 bp, encoding 683 amino acids and producing a protein of approximately 78.95 kDa. After verification, the overexpression plasmids pEGFP-AMPD1, Cas9/sgRNA2, and Cas9/sgRNA3 were found to have biological activity in chicken muscle cells and individual chickens, and two sgRNAs (sgRNA2, sgRNA3) were identified that could edit AMPD1. The qPCR and Western blotting result showed that the pEGFP-AMPD1 plasmid significantly increased both mRNA and protein expression of AMPD1. T7EI digestion showed editing efficiencies of approximately 35 %, 37 %, and 33 % for sgRNA2, sgRNA3, and sgRNA2 + sgRNA3 of AMPD1 in chicken muscle cells. In comparison, TA cloning sequencing showed editing efficiencies of approximately 36.7 %, 86.7 %, and 26.7 % and editing efficiencies in chicken individuals of approximately 71 %, 45 %, and 76.7 %, respectively. These results provide a theoretical basis and support for further investigation into the function of the AMPD1 gene.

The complete mitochondrial genome of a new species of the genus Schizothorax from Sichuan, China (Cypriniformes: Cyprinidae).

Schizothorax gulinensis sp. nov. is a new species of the genus Schizothorax from Sichuan, China (Cypriniformes: Cyprinidae). In this study, we have first reported the complete mitochondrial genome of S. gulinensis with Illumina sequencing. There were 16,587 nucleotide pairs in the mitochondrial genome (mitogenome) of S. gulinensis, including 13 protein-coding genes (PCGs), two ribosomal RNAs (rRNAs), and 22 transfer RNAs (tRNAs), as well as one non-coding control region (CR). The proportion of nucleotides in mitochondrial genome was 29.67% (A), 25.45% (T), 17.84% (G), 27.05% (C), and A + T content was 55.12%. All PCGs have the same start codon of the standard ATG, excepting for that of NADH dehydrogenase subunit 1 (nad1) which was the ATC, NADH dehydrogenase subunit 5 (nad5) which was the ATT and cytochrome c oxidase 1 (cox1) which was the ATC. Phylogenetic analysis results supported that S. gulinensis was closely related to Schizothorax grahami. The complete mitochondrial sequence of S. gulinensis will contribute to mitochondrial genome database and provide useful resources for population genetics and evolution analyses.

Dietary Methionine Increased the Growth Performances and Immune Function of Partridge Shank Broilers after Challenged with Coccidia.

The present study investigated the effects of methionine (Met) on growth, immune function, and antioxidant capacity in partridge shank broilers, which were treated with either an anticoccidial drug or a coccidia vaccine. Chickens were fed five graded levels of Met (0.33%, 0.39%, 0.45%, 0.51%, or 0.57%) for 21 days in combination with the drug or vaccine. The results revealed that an optimal level of Met supplementation (1) increased ADFI (average daily feed intake), ADG (average daily gain), and F/G values (feed-to-gain ratio), indicating improved production; (2) increased OPG levels (oocysts per gram feces), intestinal lesion scores, bursa of Fabricius and thymus indexes, and sIgA content; (3) improved GSH-Px activities, and increased content levels of T-protein, albumin, and urea nitrogen. In addition, birds in the anticoccidial drug group had higher final weights, higher ADFI and ADG values, as well as lower F/G values, compared with birds in the vaccine group, indicating that coccidia vaccine reduces the performance of broilers. In conclusion, we found that an optimal level of dietary Met improved the production of partridge shank broilers, and this result might be related to immune function and antioxidant capacity. Optimal levels of digestible Met in terms of production performance (ADG and F/G) and immune function (sIgA in ileum mucosa) in partridge shank broilers (1-21 days) were found to be 0.418, 0.451, and 0.451 of diet, respectively, when birds were given anticoccidial drug treatment, with corresponding figures of 0.444, 0.455, and 0.452% when the coccidia vaccine was administered.

Prolonged darkness attenuates imidacloprid toxicity through the brain-gut-microbiome axis in zebrafish, Danio rerio.

The present study investigated the toxic effects of IMI on brain and gut of zebrafish (Danio rerio) by a combination of transcriptome and microbiome analysis. In addition, the involvement of light/dark period was also evaluated. An acute toxic test was conducted on adult zebrafish weighing 0.45 ± 0.02 g with 4 experimental groups (n = 15): 1) IMI group (Light: Dark = 12: 12 h), 2) prolonged light group (Light: Dark = 20: 4 h), 3) prolonged darkness group (Light: Dark = 4: 20 h) which received 20 mg/L of IMI, and 4) control group, which was not treated with IMI (Light: Dark = 12: 12 h). The results showed that prolonged darkness improved the survival rate of zebrafish upon IMI exposure for 96 h. In the sub-chronic test, zebrafish were divided into the same 4 groups and exposed to IMI at 1 mg/L for 14 d (n = 30). The results showed that IMI induced oxidative stress in both IMI and prolonged light groups by inhibition of antioxidant activities and accumulation of oxidative products. Transcriptome analysis revealed a compromise of antioxidation and tryptophan metabolism pathways under IMI exposure. Several genes encoding rate-limiting enzymes in serotonin and melatonin synthesis were all inhibited in both IMI and LL groups. Meanwhile, significant decrease (P < 0.5) of serotonin and melatonin levels was observed. However, there's remarkable improvement of biochemical and transcriptional status in prolonged darkness group. In addition, microbiome analysis showed great alteration of gut bacterial community structure and inhibition of tryptophan metabolism pathway. Similarly, the gut microbiota dysbiosis induced by IMI was alleviated in prolonged darkness. In summary, sub-chronic IMI exposure induced neurotoxicity and gut toxicity in zebrafish by oxidative stress and impaired the brain-gut-axis through tryptophan metabolism perturbation. Prolonged darkness could effectively attenuate the IMI toxicity probably through maintaining a normal tryptophan metabolism.

Imidacloprid induces locomotion impairment of the freshwater crayfish, Procambarus clarkii via neurotoxicity and oxidative stress in digestive system.

Imidacloprid (IMI) is used in integrated farming like the rice-crayfish co-culture system to prevent water weevil. However, the toxic effect of IMI on the freshwater crayfish Procambarus clarkii is unknown. In the current study, the effects of IMI on the locomotion, antioxidative status, digestion and intestinal microbiota of P. clarkii were investigated. The results showed that IMI caused locomotion impairment with reduced crawl velocity, and attenuated their dark preference, aggressiveness and reversal ability. Inhibited AChE in muscle and hepatopancreas indicates the neurotoxicity of IMI which may directly lead their locomotion dysfunction. The increase of antioxidative enzymes activity and MDA level were found after 25 µg/L and 250 µg/L exposure. Significant up-regulation of several antioxidative and immune-related genes, including CZ-SOD, CAT, GPx, GST, AFL, proPO, HSP27 and HSP70 confirmed that oxidative stress was induced in all treatments when exposed to IMI. In addition, there was significant increase of LDH, indicating the different energy allocation during the exposure. Meanwhile, results from DNA damage analysis showed elevated OTM value and 8-OHdG level in hepatopancretic cells. On the other hand, decreases of alpha-amylase, lipase and increase of trypsin in hepatopancreas was observed at 25 and 250 µg/L. In addition, significant changes of composition of intestinal microbiota at both phylum and genus levels were observed according to the 16S rRNA sequencing results. Increase of pathogenic genera and decrease of beneficial bacterial communities revealed the disequilibrium of intestinal flora of crayfish. In summary, results in the present study suggest that IMI at environmentally realistic concentration could induce AChE inhibition and oxidative stress, conjointly leading the locomotion impairment in crayfish. IMI also affected the digestive functions by enzymes inhibition and gut microbiota dysbiosis.

Cytotoxicity induced by abamectin exposure in haemocytes of Chinese mitten crab, Eriocheir sinensis.

Abamectin is widely utilized in both agricultural and aquaculture areas in China for pest control. However, information about toxic effects of abamectin on non-target aquatic organisms is still incomplete. The Chinese mitten crab, Erocheir sinensis has been extensively bred in the rice-crab co-culture system for years, resulting in the frequent exposure to pesticides including abamectin. In the present study, a primary haemocyte culture model was established to investigate the immune response under exposure of abamectin. The results showed that medium osmolarity ranging from 360 to 480 mOsM/Kg was optimal for primary haemocyte culture from E. sinensis. Abamectin could induce significant decrease of cell viability, inhibition of phagocytic activity, as well as decline of acid phosphatase (ACP) and alkaline phosphatase (AKP) activities. All parameters decreased in time- and dose- dependent manners throughout the experiment, indicating the remarkable immunosuppression of abamectin on E. sinensis and also the sensitivity of the cytotoxicology model of haemocytes in vitro under abamectin exposure. In addition, a dose dependent increase of intracellular reactive oxygen species (ROS) was found after 6 h exposure. It revealed that excessive generation of ROS may a main reason to the degradation of cell viability, and moreover, the decrease in immune function.

Effects of imidacloprid on the oxidative stress, detoxification and gut microbiota of Chinese mitten crab, Eriocheir sinensis.

Imidacloprid (IMI) is used in integrated aquaculture systems for pest control and the toxicity of IMI to non-target aquatic animals such as fish and microcrustaceans has been recognised. However, knowledge about the toxic effect of IMI on commercial crabs is still scarce. In the present study, effects of IMI on the acute toxicity, antioxidative status, detoxification systems and gut microbiota in Chinese mitten crab, Erocheir sinensis were investigated. In the present study, the 96-h LC50 of IMI for E. sinensis was 24.97 mg/L. Under sublethal exposure, superoxide dismutase (SOD) activities increased under low concentration (LC, 5 µg/L) and median concentration (MC, 50 µg/L) exposure, but decreased in high concentration group (HC, 500 µg/L). Activities of catalyse (CAT) decreased in a dose-dependent manner. Detoxification-related enzymes aminopyrine N-demethylase (APND) and erythromycin N-demethylase (ERND) increased in all treatments whereas glutathione-S-transferase (GST) decreased dose-dependently. The relative mRNA expression of the cytochrome P4502 (cyp2) gene was induced significantly in LC and HC groups while no significant change was observed in cytochrome P4503 (cyp3) gene. The expression of gst was also significantly decreased in HC group. Up-regulation of heat shock protein hsp70 and 90 was observed in MC and HC groups whereas hsp60 up-regulated only in LC group. In addition, significant changes of composition of microbial communities at both phylum and genus levels were found in this test. In particular, beneficial bacteria were found to decrease and pathogens increased after exposure to IMI. These results indicate that high concentration of IMI could induce oxidative stress and suppress the detoxification system mainly by down-regulation of gst mRNA expression, inhibition of enzyme activities and dysbiosis of gut microbiota.

Abamectin at environmentally-realistic concentrations cause oxidative stress and genotoxic damage in juvenile fish (Schizothorax prenanti).

Abamectin (ABM) has been extensively used in Chinese aquaculture systems for parasite control, but no information is available regarding its effects on the important freshwater commercial fish species Schizothorax prenanti. We performed an acute toxicity test to determine the effects of ABM on S. prenanti, and the 48- and 96-h median lethal concentration values were 33.32 and 15.98 µg/L, respectively. In a second test, animals were exposed to sublethal concentrations of ABM (0.5, 2 or 8 µg/L) for 8 days, and various cytological and biochemical parameters were measured. ABM caused DNA damage in hepatocytes, with significant increases in Olive Tail Moment values and 8-hydroxy-2'-deoxyguanosine levels. Hepatocytic apoptosis occurred following all treatments, and was accompanied by an increase in reactive oxygen species (ROS) generation and caspase activity in a dose- and time-dependent manner. In addition, there were significant decreases in glutathione peroxidase levels and superoxide dismutase and catalase activity and increases in malonaldehyde levels. ABM-induced hepatocytic apoptosis in S. prenanti was probably triggered by ROS generation following a cascade reaction of caspases in mitochondrial or death receptor pathways, which caused antioxidant inhibition, oxidative product accumulation, and DNA damage in the liver.

Avermectin induces the oxidative stress, genotoxicity, and immunological responses in the Chinese Mitten Crab, Eriocheir sinensis.

Avermectin is commonly used in aquaculture systems for pest control in recent decades in China. However, no information is provided for the toxic effect to the important commercial species, Chinese mitten crab, Eriocheir sinensis. To investigate the aquatic toxicity of avermectin, an acute toxic test was performed in this study. The results showed that the 48 h- and 96 h- LC50 were 1.663 and 0.954 mg/L, respectively. For further research, crabs were exposed to sublethal concentrations of 0.03, 0.06, 0.12, 0.24 and 0.48 mg/L. Levels of antioxidants, including superoxide dismutase (SOD), catalase (CAT) and total antioxidant capacity (T-AOC) were significantly (P<0.05) decreased with dose- and time- dependent responses, meanwhile the oxidative products including malondialdehyde (MDA), hydrogen peroxide (H2O2) and protein carbonyl in serum increased significantly (P<0.05) at concentrations of 0.24 and 0.48 mg/L throughout the experiment. A significant (P<0.05) increase of intracellular ROS and decrease of phagocytic activity was observed in high concentration groups, with dose- and time- dependent manners during the exposure. In addition, serious genetic damage was detected, for the significant increase (P<0.05) of both comet ratio and %DNA in tail at each concentration, and micronucleus (MN) frequency at concentrations of 0.12, 0.24 and 0.48 mg/L at 96 h. These results indicated that sublethal concentration exposure of avermectin had a prominent toxic effect on E. sinensis based on the oxidative stress induced by generated ROS, immunological activity inhibition and genotoxicity.

Effects of deltamethrin on the antioxidant defense and heat shock protein expression in Chinese mitten crab, Eriocheir sinensis.

In the present study, the status of antioxidant response and molecular regulation in Chinese mitten crab, Eriocheir sinensis under the exposure of synthetic pyrethroid deltamethrin were investigated by means of measuring the antioxidative enzyme activity and relative mRNA expression of heat shock proteins (HSPs) in hepatopancreas. The results showed that activity of superoxide dismutase (SOD) and catalase (CAT) decreased remarkably in all treatments except the SOD activity at concentration of 0.073 µg/L. The oxidative stress products malondialdehyde (MDA) and hydrogen peroxide (H2O2) increased significantly at high concentrations while no significant difference was observed at concentrations of 0.073 and 0.146 µg/L throughout the experiment. Meanwhile, the relative mRNA expression of HSP 60, HSP 70 and HSP 90 was significantly up-regulated in all treatments at each time point. All resutls above indicated that deltamethrin has prominent toxic effect on E. sinensis based on antioxidative enzyme inhibition and oxidative products accumulation at environmental related concentrations, and a protective response by up-regulation of HSPs was carried out by animals to mitigate the oxidative stress. In addition, SOD, CAT, MDA, H2O2 and the expression of heat shock proteins, especially HSP 70 in hepatopancreas could be sensitive biomarkers in the assessment of toxic effect of deltamethrin on E.sinensis.

Clinically isolated enterovirus A71 subgenogroup C4 strain with lethal pathogenicity in 14-day-old mice and the application as an EV-A71 mouse infection model.

The Enterovirus A71 (EV-A71) subgenogroup C4 is prevalent in China. EV-A71 causes hand, foot and mouth disease (HFMD) in children and may lead to severe neurological diseases. The development of antiviral and protective vaccines against EV-A71 is significantly hindered by the lack of suitable animal models to recapitulate human neurological symptoms. In this study, GZ-CII, a highly virulent EV-A71 subgenogroup C4 strain, was isolated from hospitalized children with HFMD. Intraperitoneal infections of GZ-CII resulted in progressive neurological disease in mice as old as 14 days. Administration of an inactivated EV-A71 vaccine or an anti-EV-A71 immune serum protected the mice against the GZ-CII infection. This demonstrated that a mouse model with EV-A71 GZ-CII could be used to evaluate potential vaccine candidates and therapeutics for subgenogroup C4. Comparing the genome sequence of GZ-CII with that of the avirulent EV-A71 subgenogroup C4 strain revealed unique mutations in GZ-CII. When mutation VP2-K149I was introduced into the nonpathogenic EV-A71 subgenogroup C4 strain, the variant similar to GZ-CII significantly increased viral replication and virulence in mice. These results indicated that the VP2-K149I mutation played an important role in enhancing the virulence of the EV-A71 subgenogroup C4 strain in mice, and that mice infected with the GZ-CII strain are a promising model for evaluating vaccines and therapeutics against the EV-A71 subgenogroup C4.