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Crucian carp (Carassius auratus gibelio), an extensively cultivated freshwater fish, was one of the model species for the study of fish immunology. Polyclonal antibodies were advantageous molecular tools for studying teleost immune system. Specifically, polyclonal antibodies reacting with immunoglobulins (Ig) were used successfully in studies of the teleost fishes. In the present study, we produced polyclonal antibody against CH2 domains of crucian carp IgM, and measured the in vivo dynamics of IgM mRNA caused by CyHV-2 infection. The recombinant protein IgM with relative molecular weight about 53 KD was correctly expressed in prokaryotic cells. The specificity of the polyclonal antibody was evaluated by Western blotting and results revealed that the antibody not only specifically recognized crucian carp serum but also cross-reacted with grass carp serum. Quantitative RT-PCR analysis demonstrated the expression of IgM mRNA changed significantly after CyHV-2 infection. The expression of IgM in the kidney increased and reached a maximum at 6 h post-infection (hpi), while dropped to a low level at 5 days post-infection (dpi). In conclusion, the expression of IgM was significantly upregulated in the kidney of crucian carp infected with CyHV-2, indicating that IgM played a potential role in systemic immunity against viral infection. Polyclonal antibody against crucian carp IgM had certain clinical relevance, which might provide insight into the early stage of virus infection and prevention of the disease.

Cyprinid herpesvirus 2 (CyHV-2) is the pathogen of herpesviral hematopoietic necrosis (HVHN), causing the severe economic losses in farmed gibel carp (Carassius gibelio). Further exploration of the genome structure and potential molecular pathogenesis of CyHV-2 through complete genome sequencing, comparative genomics, and molecular characterization is required. Herein, the genome of a CyHV-2 YC-01 strain isolated from diseased gibel carp collected in Yancheng, Jiangsu Province, China was sequenced, then we analyzed the genomic structure, genetic properties, and molecular characterization. First, the complete YC-01 genome comprises 275,367 bp without terminal repeat (TR) regions, with 151 potential open reading frames (ORFs). Second, compared with other representative published strains of the genus Cyvirus, several evident variations are found in YC-01, particularly the orientation and position of ORF25 and ORF25B. ORF107 and ORF156 are considered as potential molecular genetic markers for YC-01. ORF55 (encoding thymidine kinase) might be used to distinguish YC-01 and ST-J1 from other CyHV-2 isolates. Third, phylogenetically, YC-01 clusters with the members of the genus Cyvirus (together with the other six CyHV-2 isolates). Fourth, 43 putative proteins are predicted to be functional and are mainly divided into five categories. Several conserved motifs are found in nucleotide, amino acid, and promoter sequences including cis-acting elements identification of YC-01. Finally, the potential virulence factors and linear B cell epitopes of CyHV-2 are predicted to supply possibilities for designing novel vaccines rationally. Our results provide insights for further understanding genomic structure, genetic evolution, and potential molecular mechanisms of CyHV-2.

The mode and outcome of fish–virus interactions are influenced by many abiotic factors, among which water temperature is especially important in poikilothermic fish. Rare minnow Gobiocypris rarus is a eurythermal small cyprinid fish that is sensitive to infection with genotype II grass carp reovirus (GCRV). HSP70, a conservative and key player in heat shock response, is previously identified as an induced pro-viral factor during GCRV infection in vitro. Here, rare minnow was subjected to heat shock treatment (HST), 1 h treatment at 32 °C followed by reverting to a normal temperature of 24 °C, and subsequently challenged with GCRV-II at a dosage of 1 × LD50. The effect of HST on GCRV virulence in vivo was evaluated by calculating virus-associated mortality and viral load in both dead and survival fish. The results revealed that HST enhanced the mortality of rare minnow infected with GCRV; the fact that viral loads in the tissue samples of HST-treated fish were significantly higher than those in samples of the control group at 6, 8 d p.i. reflected a faster infection process due to HST. Quantitative gene expression analysis was further employed to show that the expression levels of Hsp70 in intestine and liver tissues from the HST group declined faster than muscle tissue after HST. HST W/O GCRV challenge upregulated proinflammatory cytokines such as MyD88 and Nf-κB, which was in consistence with the inflammation observed in histopathological analysis. This study shed light on the complexity of the interaction between fish abiotic and biotic stress response, which suggested that HST, an abiotic stress, could enhance the virulence of GCRV in Gobiocypris rarus that involved modulating the gene expression of host heat shock, as well as a pro-inflammatory response.

Hydrophobic interactions are widely applied in diverse fields, yet the effect of the degree of hydrophobicity on the interactions between hydrophobic surfaces is still unaddressed, limiting the development of functional hydrophobic surfaces. In this study, the interfacial interactions and configuration of hydrophobic chains on surfaces with ranging hydrophobicity are surveyed by surface forces apparatus. Our findings revealed that with partial hydrophobicity, a long-range electrical double-layer repulsive force pertains between surfaces, while between fully hydrophobized surfaces, only steric hindrance is observed, and the hydrophobic chains maintain configuration even after long time compression. Our findings provide useful implications for understanding hydrophobic interactions.

Abstract Background Cyprinid herpesvirus 2 (CyHV-2) is a pathogenic fish virus belonging to family Alloherpesviridae. The CyHV-2 gene encoding thymidine kinase (TK) is an important virulence-associated factor. Therefore, we aimed to investigate the biological function of open reading frame 55 (ORF55) in viral replication. Methods Purified CyHV-2 ORF55 protein was obtained by prokaryotic expression, and the interacting peptide was screened out using phage display. Host interacting proteins were then predicted and validated. Results ORF55 was efficiently expressed in the prokaryotic expression system. Protein and peptide interaction prediction and dot-blot overlay assay confirmed that peptides identified by phage display could interact with the ORF55 protein. Comparing the peptides to the National Center for Biotechnology Information database revealed four potential interacting proteins. Reverse transcription quantitative PCR results demonstrated high expression of an actin-binding Rho-activating protein in the latter stages of virus-infected cells, and molecular docking, cell transfection and coimmunoprecipitation experiments confirmed that it interacted with the ORF55 protein. Conclusion During viral infection, the ORF55 protein exerts its biological function through interactions with host proteins. The specific mechanisms remain to be further explored.

Hydration plays a crucial role in cement composites, but the traditional methods for measuring hydration heat face several limitations. In this study, we propose a machine learning-based approach to predict hydration heat at specific time points for three types of cement composites: ordinary Portland cement pastes, fly ash cement pastes, and fly ash–metakaolin cement composites. By adjusting the model architecture and analyzing the datasets, we demonstrate that the optimized artificial neural network model not only performs well during the learning process but also accurately predicts hydration heat for various cement composites from an extra dataset. This approach offers a more efficient way to measure hydration heat for cement composites, reducing the need for labor- and time-intensive sample preparation and testing. Furthermore, it opens up possibilities for applying similar machine learning approaches to predict other properties of cement composites, contributing to efficient cement research and production.

Cyprinid herpesvirus 2 (CyHV-2) is a pathogen that causes significant losses to the global aquaculture industry due to mass mortality in crucian carp and goldfish. This study demonstrates that the ORF55/ORF57 deletion mutants CyHV-2-Δ55-CP and CyHV-2-Δ57-CP obtained through homologous recombination replicate effectively within the caudal fin of Carassius auratus gibelio (GiCF) cells and exhibit morphologies similar to the CyHV-2 wild-type strain. Both mutants demonstrated a decrease in virulence, with CyHV-2-Δ57-CP exhibiting a more significant reduction. This serves as a reference for the subsequent development of recombinant attenuated vaccines against CyHV-2. Additionally, both mutants expressed the inserted RGNNV-CP (capsid protein of Redspotted grouper nervous necrosis virus) fusion protein gene, and inoculation with CyHV-2-Δ57-CP-infected GiCF cell lysates elicited an antibody response in the grouper. These results indicate that, while ORF55 and ORF57 genes of CyHV-2 are not required for viral replication in vitro, they do play a role in virulence in vivo. Additionally, expression of foreign protein in CyHV-2 suggests that the fully attenuated mutant of CyHV-2 could potentially function as a viral vector for developing subunit vaccines or multivalent recombinant attenuated vaccines.

How good dispersion of nano-SiO2 (NS) plays a great influence on how strong reinforcing efficiency to cement-based materials. In this study, a novel nanohybrid of polycarboxylate ether (PCE) grafted nano-silica sol (PCE@NSS) was prepared by sol–gel methods to chemically adsorb PCE onto NSS. The developed nanohybrid showed a higher ultraviolet–visible (UV–Vis) absorbance and slower sedimentation in alkaline solutions, indicating the better dispersion and stability of PCE@NSS than that of untreated NSS. By this way, only 0.05 wt.% addition of PCE@NSS was helpful to refine the capillary pore size and reduce the total porosity of cement mortar, which in turn improved the compressive and flexural strength of cement mortar by 18.34% and 10.5% at 28 days, respectively. In addition, a reinforcing index (R index) with the ratio of strength enhancement % to dosage % was proposed, indicating that the R index of PCE@NSS showed 10 times higher than that of other NS-based nanomaterials reported in literatures, addressing the highly reinforcing efficiency of PCE@NSS to cement mortar. Furthermore, water absorption coefficient of cement mortar was therefore reduced by 12.7% with 0.2 wt.% addition of PCE@NSS. In conclusion, such an ultra-low dosage of PCE@NSS with better dispersion and stability exhibited great potential to reinforce cement-based materials with highly efficiency, lower cost and reduced carbon emissions.

IntroductionRibonucleotide reductase (RR) is essential for the replication of the double-stranded DNA virus CyHV-2 due to its ability to catalyze the conversion of ribonucleotides to deoxyribonucleotides, and is a potential target for the development of antiviral drugs to control CyHV-2 infection.MethodsBioinformatic analysis was conducted to identify potential homologues of RR in CyHV-2. The transcription and translation levels of ORF23 and ORF141, which showed high homology to RR, were measured during CyHV-2 replication in GICF. Co-localization experiments and immunoprecipitation were performed to investigate the interaction between ORF23 and ORF141. siRNA interference experiments were conducted to evaluate the effect of silencing ORF23 and ORF141 on CyHV-2 replication. The inhibitory effect of hydroxyurea, a nucleotide reductase inhibitor, on CyHV-2 replication in GICF cells and RR enzymatic activity in vitro was also evaluated.ResultsORF23 and ORF141 were identified as potential viral ribonucleotide reductase homologues in CyHV-2, and their transcription and translation levels increased with CyHV-2 replication. Co-localization experiments and immunoprecipitation suggested an interaction between the two proteins. Simultaneous silencing of ORF23 and ORF141 effectively inhibited the replication of CyHV-2. Additionally, hydroxyurea inhibited the replication of CyHV-2 in GICF cells and the in vitro enzymatic activity of RR.ConclusionThese results suggest that the CyHV-2 proteins ORF23 and ORF141 function as viral ribonucleotide reductase and their function makes an effect to CyHV-2 replication. Targeting ribonucleotide reductase could be a crucial strategy for developing new antiviral drugs against CyHV-2 and other herpesviruses.

Activating protein-1 (AP-1) composed of Jun and Fos family proteins, which are frequently activated by pathogens to induce cellular inflammation and immune responses. In this study, by using RNA sequencing (RNA-seq) technology, we found that FosB, Fosl2, and JunD expression was co-induced 12 and 20 h in grass carp (Ctenopharyngodon idella) kidney cell line (CIK cells) after grass carp reovirus (GCRV) infection. Full-length cDNA of the three genes were cloned and phylogenetic analysis indicated that the corresponding proteins shared high homology with those in higher vertebrates. The tissue distribution analysis of the three genes showed they were ubiquitously expressed in various tissues of healthy grass carp. In infection assay, FosB, Fosl2, and JunD were up-regulated and co-expressed 8 h after GCRV infection in CIK cells. Protein interaction and co-expression network analysis revealed that FosB, Fosl2, and JunD showed similar functions and were co-expressed in mammals and fish. Taken together, these results indicated that the co-expression of the AP-1 family proteins in grass carp uniformly responds to GCRV infection. These findings provided insights into the further study of AP-1 transcription factors function during grass carp reovirus infection.

Abstract In order to research the ability of AGV to cope with the dynamic path planning in mobile single-object dynamic environment, the dynamic path planning methods based on reduced point and non-reduced point were proposed. This paper focuses on the dynamic path planning method of non-reduced point moving targets, adopts the method of combining historical data and real-time data, establishing the NAR neural network prediction model to predict velocity of moving target, and connecting with the A * algorithm, through the example simulation experiment, the results show that the effective success rate of the algorithm reaches 77.5% in the map environment set in this paper.

The suspension system’s performance has a direct impact on the ride comfort, handling stability, and driving safety of heavy vehicles. The active air suspension of heavy vehicles can adjust the stiffness, damping parameters, and body height in real-time based on different road conditions. This adjustment ensures that the entire vehicle experiences a smooth ride while also making vehicle loading and unloading more labor efficient. Additionally, the active air suspension system enables the vehicle to achieve automation and intelligence. This study focused on a particular 6 × 4 heavy vehicle and designed an active air suspension system that aligns with the vehicle parameters of the heavy truck. Through the use of a fuzzy PID active control strategy, this study investigated and analyzed the height adjustment of the air spring. The results indicate that at a vehicle speed of 60 km/h on a class A road surface, the vehicle body’s vertical acceleration was reduced by 22.1%, and the dynamic travel of the suspension was reduced by 20.1%. This indicates that the fuzzy PID active air suspension system effectively reduces the vehicle’s vibrations and improves ride comfort.

Cyprinid herpesvirus 2 (CyHV-2) has caused great economic loss to the crucian carp breeding industry. Upon viral stimulation, eukaryotic cells generally activate the expression of anti-oxidative genes to maintain the intracellular oxidative balance and resist viral infection. Here, intracellular reactive oxygen species (ROS) levels in CyHV-2-infected cells were monitored to show that CyHV-2 induced the increase of intracellular ROS during early infection, and intracellular excessive accumulation of ROS was ameliorated during late infection, which was accompanied by activated expression of genes related to Nrf2 signaling pathway. In order to explore the interaction between CyHV-2 infection and ROS production, RyuF-2 cells were treated with either antioxidant epigallocatechin-3-gallate (EGCG) or berberine hydrochloride (BBH) and then infected with CyHV-2. Both BBH and EGCG could effectively inhibit the amplification of CyHV-2 while inhibiting the accumulation of intracellular ROS. Consistent with this, the oxidant stress-related genes were up-regulated by CyHV-2 infection and down-regulated in cells treated with either BBH or EGCG, through which the production of intracellular ROS was modulated. These results collectively demonstrated that early ROS accumulation favored the replication of CyHV-2, while antioxidants (BBH and EGCG) could inhibit the amplification of CyHV-2 by inhibiting ROS induction.

Cyprinid herpesvirus 2 (CyHV-2) is a causative factor of herpesviral hematopoietic necrosis (HVHN) in farmed crucian carp (Carassius carassius) and goldfish (Carassius auratus). In this study, we analyzed the genomic characteristics of a new strain, CyHV-2 SH-01, isolated during outbreaks in crucian carp at a local fish farm near Shanghai, China. CyHV-2 SH-01 exhibited a high sensitivity to goldfish and crucian carp in our previous research. The complete genome of SH-01 is 290,428 bp with 154 potential open reading frames (ORFs) and terminal repeat (TR) regions at both ends. Compared to the sequenced genomes of other CyHVs, Carassius auratus herpesvirus (CaHV) and Anguillid herpesvirus 1 (AngHV-1), several variations were found in SH-01, including nucleotide mutations, deletions, and insertions, as well as gene duplications, rearrangements, and horizontal transfers. Overall, the genome of SH-01 shares 99.60% of its identity with that of ST-J1. Genomic collinearity analysis showed that SH-01 has a high degree of collinearity with another three CyHV-2 isolates, and it is generally closely related to CaHV, CyHV-1, and CyHV-3, although it contains many differences in locally collinear blocks (LCBs). The lowest degree of collinearity was found with AngHV-1, despite some homologous LCBs, indicating that they are evolutionarily the most distantly related. The results provide new clues to better understand the CyHV-2 genome through sequencing and sequence mining.

The hydraulic accumulator has the advantages of high power density, fast response, stable operation and high cost performance. However, compared with the electric energy storage method, the hydraulic accumulator has low energy density and large pressure fluctuation while absorbing and discharging energy, which severely limits its application in hydraulic excavators. To improve the potential energy loss of the boom during the lowering process, an electro-hydraulic drive and energy recovery system for excavator booms (EHDR-EEB) based on a battery and accumulator is proposed. As a result, a simulation model of the electro-hydraulic drive and energy management strategy of a 1.6 t pure electric hydraulic excavator is built to investigate the energy regeneration and utilization. The simulation outcomes show that the potential energy recovery rate is as high as 92%. This research on EHDR-EEB makes a significant contribution to the economic improvement of electric hydraulic excavators.

Neomycin belongs to the family of 2-deoxystreptamine-containing aminoglycoside antibiotics. It is widely used for bacterial infections, targeting most gram-negative and some gram-positive bacteria. Neomycin has also been reported to show antiviral activity. Here, we evaluated the toxicity of neomycin sulfate, and investigated its effect on non-specific immunity and viral infection in crucian carp (Carassius auratus gibelio). The safe concentration of neomycin sulfate for crucian carp was determined to be 102.9 mg/kg in vivo. In oxidative damage assays, neomycin sulfate increased superoxide dismutase and catalase activity and decreased malondialdehyde in the liver of crucian carp. In non-specific blood immune assays, the white blood cell count and complement 3 content significantly increased after neomycin sulfate treatment, while no significant changes were observed in antibacterial or lysozyme activity. In a challenge test, neomycin sulfate protected the crucian carp from cyprinid herpesvirus 2 (CyHV-2) infection and inhibited CyHV-2 replication. In cytotoxicity assays, low concentrations of neomycin sulfate had no cytotoxicity on cells from the fins of crucian carp. The results of the present study indicate that oral administration of neomycin sulfate reduced oxidative damage, enhanced immunity and provided protection against CyHV-2 in crucian carp. Keywords: Neomycin sulfate, Non-specific immunity, Oxidative damage, Crucian carp, Cyprinid herpesvirus 2

DNA viruses, most notably members of the herpesvirus family, generally encode miRNAs to mediate both virus and host genes expression. We previously demonstrated that Cyprinid herpesvirus 2 (CyHV-2) encodes 17 miRNAs that are involved in innate immune signaling pathways. In this study, the function of CyHV-2-encoded miRNA was further investigated in GiCF cells. We found that miR-C4 promoted CyHV-2-induced apoptosis, while miR-C12 decreased CyHV-2-induced apoptosis. miR-C12 targeted to 3′ UTR sequence of caspase 8 and suppressed the expression of caspase 8. Besides, the silencing of caspase 8 by specific siRNA led to the attenuation of CyHV-2-induced apoptosis. Furthermore, caspase 8 was downregulated in cells transfected with miR-C12 during CyHV-2 infection. Overexpression of miR-C12 significantly suppressed CyHV-2-induced apoptosis, while silencing of miR-C12 promoted CyHV-2-induced apoptosis. Finally, inhibition of miR-C12 resulted in suppression of CyHV-2 propagation, overexpression of miR-C12, and CASP8-siRNA-1 facilitated CyHV-2 propagation. Taken together, our results demonstrated that CyHV-2-encoded miR-C12 to suppress virus-induced apoptosis and promoted virus replication by targeting caspase 8.

Abstract Background Grass carp (Ctenopharyngodon idella) hemorrhagic disease is caused by an acute infection with grass carp reovirus (GCRV). The frequent outbreaks of this disease have suppressed development of the grass carp farming industry. GCRV104, the representative strain of genotype III grass carp (Ctenopharyngodon idella) reovirus, belongs to the Spinareovirinae subfamily and serves as a model for studying the strain of GCRV which encodes an outer-fiber protein. There is no commercially available vaccine for this genotype of GCRV. Therefore, the discovery of new inhibitors for genotype III of GCRV will be clinically beneficial. In addition, the mechanism of GCRV with fiber entry into cells remains poorly understood. Methods Viral entry was determined by a combination of specific pharmacological inhibitors, transmission electron microscopy, and real-time quantitative PCR. Results Our results demonstrate that both GCRV-JX01 (genotype I) and GCRV104 (genotype III) of GCRV propagated in the grass carp kidney cell line (CIK) with a typical cytopathic effect (CPE). However, GCRV104 replicated slower than GCRV-JX01 in CIK cells. The titer of GCRV-JX01 was 1000 times higher than GCRV104 at 24 h post-infection. We reveal that ammonium chloride, dynasore, pistop2, chlorpromazine, and rottlerin inhibit viral entrance and infection, but not nystatin, methyl-β-cyclodextrin, IPA-3, amiloride, bafilomycin A1, nocodazole, and latrunculin B. Furthermore, GCRV104 and GCRV-JX01 infection of CIK cells depended on dynamin and the acidification of the endosome. This was evident by the significant inhibition following prophylactic treatment with the lysosomotropic drug ammonium chloride or dynasore. Conclusions Taken together, our data have suggested that GCRV104 enters CIK cells through clathrin-mediated endocytosis in a pH-dependent manner. We also suggest that dynamin is critical for efficient viral entry. Additionally, the phosphatidylinositol 3-kinase inhibitor wortmannin and the protein kinase C inhibitor rottlerin block GCRV104 cell entry and replication.

Cyprinid herpesvirus 2 (CyHV-2), a member of the Alloherpesviridae family belonging to the genus Cyprinivirus, is a fatal contagious aquatic pathogen that affects goldfish (Carassius auratus) and crucian carp (Carassius carassius). Although crucian carp and goldfish belong to the genus Carassius, it is unclear whether they are susceptible to the same CyHV-2 isolate. In addition, the origin of the crucian carp-derived CyHV-2 virus isolate remains unclear. CyHV-2 SH01 was isolated during herpesviral hematopoietic necrosis disease (HVHN) outbreaks in crucian carp at a local fish farm near Shanghai. CyHV-2 SH01 was confirmed by PCR and Western blot analysis of kidney, spleen, muscle, and blood tissue from the diseased crucian carp. Moreover, histopathological and ultra-pathological analyses revealed pathological changes characteristic of CyHV-2 SH01 infection in the tissues of the diseased crucian carp. In the present study, goldfish and crucian carp were challenged with CyHV-2 SH01 to elucidate viral virulence. We found that CyHV-2 SH01 could cause rapid and fatal disease progression in goldfish and crucian carp 24 h post-injection at 28 °C. Experimental infection of goldfish by injection indicated that the average virus titer in the kidney of the goldfish was 103.47 to 103.59 copies/mg. In addition, tissues exhibited the most prominent histopathological changes (cellular wrinkling and shrinkage, cytoplasmic vacuolation, fusion of the gill lamellae, and hepatic congestion) in CyHV-2 SH01-infected goldfish and crucian carp. Thus, crucian carp and goldfish showed a high sensitivity, with typical symptoms, to HVHN disease caused by CyHV-2 SH01.

Cyprinid herpesvirus 2 (CyHV-2), which infects goldfish and crucian carp causing high mortality, is an emerging viral pathogen worldwide. The genome of CyHV-2 is large and comprises double-stranded DNA, including several genes similar to cyprinid herpesvirus 1, ictalurid herpesvirus-1, cyprinid herpesvirus 3, and ranid herpesvirus-1. Genes of DNA viruses are expressed in three temporal phases: immediate-early (IE), early (E), and late (L) genes. Viral IE genes initiate transcription as soon as the virus enters the host, without viral DNA replication. IE gene products enable the efficient expression of E and L genes or regulate the host to initiate virus replication. In the present study, five IE genes of CyHV-2 were identified, including open reading frame (ORF)54, ORF121, ORF141, ORF147, and ORF155. Time course analysis and reverse transcription polymerase chain reaction confirmed five IE genes, thirty-four E genes, and thirty-nine L genes. In addition, all 150 ORFs identified in the CyHV-2 genome are transcribed, and are expressed in chronological order, similar to other herpesviruses. This study is the first to identify the IE genes of CyHV-2, which will provide more information for viral molecular characterization.

Objective: To determine factors associated with late preterm births in an underdeveloped region of China, and search for relevant reports in other underdeveloped regions by a systematic review. Materials and methods: Data of births occurring between January 2004 and December 2008 from eight hospitals in Western Sichuan Province, China, were analyzed. Late preterm birth was defined as delivery at 34–36 6/7 weeks' gestation. Medline, Cochrane Library, and Google Scholar were searched for studies which reported the risk factors of late preterm births in undeveloped regions until January 31, 2014. Results: During the study period there were 4711 late preterm births and 54,574 term births. The odds ratios (ORs) for a late preterm birth of mothers

Trichlorfon is a most widely used organophosphate insecticide in aquaculture, many successful results have been reported for bath treatments of trichlorfon to control parasites. However, immersion treatments of large stocks with trichlorfon has caused serious environmental pollution. In contrast, oral administration treatment has advantages on reducing environmental pollution and having little effect in non-targeted species. The aim of this study was to investigate the effect of trichlorfon on Carassius auratus gibelio physiology after a single oral administration. In this study, Carassius auratus gibelio was subjected to oral gavage with various concentrations of trichlorfon (0.5 g/kg, 1 g/kg, and 2 g/kg). The trichlorfon concentration in the plasma and liver tissue was quantified using liquid chromatography-tandem mass spectrometry at different time points. At the beginning of oral exposure, the uptake of trichlorfon in the plasma and liver tissue was fast, and trichlorfon was rapidly eliminated to a low level within 24 h. In addition, acetylcholinesterase, superoxide dismutase, catalase, and glutathione-S-transferase activities in the plasma and liver tissue changed significantly after trichlorfon exposure. Additionally, vacuolar degeneration, necrosis, and congestion of the central vein were observed in the liver after trichlorfon exposure, as assessed by hematoxylin and eosin staining. Our results suggested that trichlorfon could accumulate and induce hematotoxicity and hepatotoxicity in the plasma and liver tissue, the toxicity induced by trichlorfon might result in physiological disturbances in fish.

Silicon oxide-based memristors have been extensively studied due to their compatibility with the dominant silicon complementary metal−oxide−semiconductor (CMOS) fabrication technology. However, the variability of resistance switching (RS) parameters is one of the major challenges for commercialization applications. Owing to the filamentary nature of most RS devices, the variability of RS parameters can be reduced by doping in the RS region, where conductive filaments (CFs) can grow along the locations of impurities. In this work, we have successfully obtained RS characteristics in Pt dispersed silicon oxide-based memristors. The RS variabilities and mechanisms have been analyzed by screening the statistical data into different resistance ranges, and the distributions are shown to be compatible with a Weibull distribution. Additionally, a quantum points contact (QPC) model has been validated to account for the conductive mechanism and further sheds light on the evolution of the CFs during RS processes.

IE0 and IE1 are transactivator proteins of the most studied baculovirus, the Autographa californica multiple nucleopolyhedrovirus (AcMNPV). IE0 is a 72.6 kDa protein identical to IE1 with the exception of its 54 N-terminal amino acid residues. To gain some insight about important structural motifs of IE0, we expressed the protein and C‑terminal mutants of it under the control of the Drosophila heat shock promoter and studied the transactivation and replication functions of the transiently expressed proteins. IE0 was able to promote replication of a plasmid bearing the hr5 origin of replication of AcMNPV in transient transfections with a battery of eight plasmids expressing the AcMNPV genes dnapol, helicase, lef-1, lef-2, lef-3, p35, ie-2 and lef-7. IE0 transactivated expression of the baculovirus 39K promoter. Both functions of replication and transactivation were lost after introduction of selected mutations at the basic domain II and helix-loop-helix conserved structural motifs in the C-terminus of the protein. These IE0 mutants were unable to translocate to the cell nucleus. Our results point out the important role of some structural conserved motifs to the proper functioning of IE0.

BACKGROUND: De novo transcriptome sequencing is a robust method of predicting miRNA target genes, especially for organisms without reference genomes. Differentially expressed miRNAs had been identified previously in kidney samples collected from susceptible and resistant grass carp (Ctenopharyngodon idella) affected by Aeromonas hydrophila. Target identification for these differentially expressed miRNAs poses a major challenge in this non-model organism. RESULTS: Two cDNA libraries constructed from mRNAs of susceptible and resistant C. idella were sequenced by Illumina Hiseq 2000 technology. A total of more than 100 million reads were generated and de novo assembled into 199,593 transcripts which were further extensively annotated by comparing their sequences to different protein databases. Biochemical pathways were predicted from these transcript sequences. A BLASTx analysis against a non-redundant protein database revealed that 61,373 unigenes coded for 28,311 annotated proteins. Two cDNA libraries from susceptible and resistant samples showed that 721 unigenes were expressed at significantly different levels; 475 were significantly up-regulated and 246 were significantly down-regulated in the SG samples compared to the RG samples. The computational prediction of miRNA targets from these differentially expressed genes identified 188 unigenes as the targets of 5 conserved and 4 putative novel miRNA families. CONCLUSION: This study demonstrates the feasibility of identifying miRNA targets by transcriptome analysis. The transcriptome assembly data represent a substantial increase in the genomic resources available for C. idella and will provide insights into the gene expression profile analysis and the miRNA function annotations in further studies.

Cyprinid herpesvirus 2 (CyHV-2), a member of the Alloherpesviridae family belonging to the genus Cyprinivirus, was initially isolated from goldfish (Carassius auratus) and has been recently emerging as a virulent pathogen for cultured prussian carp (Carassius auratus gibelio) world-wide. In this study, a novel and effective method for concentration and quantification of live CyHV-2 virions from water was successfully established through coupling the iron flocculation with real time qPCR assay. Then, the shedding intensity of CyHV-2 in fish-tank water from artificially-challenged goldfish (25 fish/20 L) was monitored continuously for 7 days on a daily basis through quantitating viral genomic copy numbers by qPCR, and the maximum shedding level was determined to be 105 copies/L. Horizontal transmission research system was established by inoculating healthy goldfish in water spiked with serial dilution of CyHV-2 virions ranging from 107 to 10³ copies/L. Our results indicated that water-borne CyHV-2 efficiently caused the infection of tested goldfish even in a concentration of 10³ copies/L, and the overall transmission efficacy was not linearly correlated with the level of input virus in the fish tank. Commercial disinfectant Composite Sodium Chlorite Powder (CSCP) has been widely applied in aquaculture to control microbial infection through direct spill in the water, and its effect in inactivating the CyHV-2 infectivity remains unknown. We further determined that the EC50 of CSCP against 3.89 TCID50/mL CyHV-2 was close to 15.625 µg/mL in vitro, and application of CSCP in a level as high as 60 µg/mL (the safety concentration of CSCP for goldfish) couldn't protect goldfish from CyHV-2 challenge through immersion. Thus, the disinfectant CSCP was regarded as none-effective for blocking CyHV-2 transmission in water during epidemic. Overall, our data provided quantitative data to demonstrate the shedding intensity of CyHV-2 in water, and CSCP was shown to be not effective in blocking water-borne horizontal transmission of CyHV-2 in goldfish. The virus-concentration protocol and virus-inhibition assay established here also paved the way for evaluating more commercial disinfectants in their effects in blocking water-borne horizontal transmission of CyHV-2. [ABSTRACT FROM AUTHOR]