Polymeric membrane potentiometric sensors based on template-removal-free imprinted receptors for determination of antibiotics.

Currently, Nernstian-response-based polymeric membrane potentiometric sensors using molecularly imprinted polymers (MIPs) as receptors have been successfully developed for determination of organic ionic species. However, the preparation of these MIP receptors usually involves tedious and time-consuming template-removal procedures. Herein, a template-removal-free MIP is proposed and used as a receptor for fabrication of a potentiometric sensor. The proposed methodology not only significantly shortens the preparation time of MIP-based potentiometric sensors but also improves the batch-to-batch reproducibility of these sensors. By using antibiotic vancomycin as a model, the new concept offers a linear concentration range of 1.0 × 10-7 to 1.0 × 10-4 mol L-1 with a detection limit of 2.51 × 10-8 mol L-1. It can be expected that the template-removal-free MIP-based sensing strategy could lay the foundation for simple fabrication of electrochemical sensors without the need for template removal such as potentiometric and capacitive sensors and ion-sensitive field-effect transistors.

Chronopotentiometric Nanopore Sensor Based on a Stimulus-Responsive Molecularly Imprinted Polymer for Label-Free Dual-Biomarker Detection.

The development of sensors for detection of biomarkers exhibits an exciting potential in diagnosis of diseases. Herein, we propose a novel electrochemical sensing strategy for label-free dual-biomarker detection, which is based on the combination of stimulus-responsive molecularly imprinted polymer (MIP)-modified nanopores and a polymeric membrane chronopotentiometric sensor. The ion fluxes galvanostatically imposed on the sensing membrane surface can be blocked by the recognition reaction between the target biomarker in the sample solution and the stimulus-responsive MIP receptor in the nanopores, thus causing a potential change. By using two external stimuli (i.e., pH and temperature), the recognition abilities of the stimulus-responsive MIP receptor can be effectively modulated so that dual-biomarker label-free chronopotentiometric detection can be achieved. Using alpha fetoprotein (AFP) and prostate-specific antigen (PSA) as model biomarkers, the proposed sensor offers detection limits of 0.17 and 0.42 ng/mL for AFP and PSA, respectively.

Maintenance-free antifouling polymeric membrane potentiometric sensors based on self-polishing coatings.

Polymeric membrane ion-selective electrodes (ISEs) have been widely used in environmental monitoring. However, in complicated marine environments, marine biofouling usually becomes a sticky problem for these electrodes. Herein, for the first time, a novel maintenance-free antifouling potentiometric marine sensor based on a self-polishing coating (SPC) is proposed. The SPC is synthesized by using the seeded emulsion polymerization method based on the triisopropylsilyl methacrylate monomer as the regulator of the self-renewal rate. This coating can be simply modified onto the electrode surface by drop-casting. The silyl acrylate side groups of the obtained SPC on the sensor surface can be hydrolyzed in the marine alkaline medium. The shear movement of seawater driven by sea waves, wind, gravity, or vibration removes the leftover (fouled) brittle polymer backbone and thus the fouling marine microorganisms. As a proof-of-concept experiment, a polymeric membrane Ca2+-ISE is chosen as a model. Compared to the unmodified electrode, the SPC-coated Ca2+-ISE exhibits remarkable improved antifouling properties in terms of superior anti-adhesive abilities towards marine microorganisms, such as bacterial cells and algae and excellent long-term stability even in the presence of high levels of marine microorganisms. Since no additional manual maintenance is required for maintaining the antifouling abilities of the sensor, the proposed self-polishing sensor may lay an important foundation for construction of unattended long-term potentiometric monitoring systems in real marine environments.

Improvement of the selectivity of a molecularly imprinted polymer-based potentiometric sensor by using a specific functional monomer.

Potentiometric sensors based on the molecularly imprinted polymers (MIPs) as the receptors have been successfully developed for determination of various organic and biological species. However, these MIP receptors may suffer from problems of low selectivity. Especially, it would be difficult to distinguish the target analyte from its structurally similar interferents. In this work, we propose a novel strategy that using specific functional monomer to fabricate MIP with high selectivity towards the target molecule. The density functional theory calculations are used to investigate the interactions between the template and the functional monomer. The binding energy between the template and functional monomer can be used as the criterion for identifying the optimal monomer. As a proof-of-concept experiment, bisphenol A (BPA) is chosen as the template and the MIP is synthesized by the precipitation polymerization method using the specific allyl-ß-cyclodextrin (allyl-ß-CD) with high affinity towards BPA as the functional monomer. The high-affinity MIP is employed as the receptor for the construction of the potentiometric sensor. The proposed potentiometric sensor based on the MIP using allyl-ß-CD as the functional monomer shows an improved response performance in terms of selectivity and sensitivity compared to the conventional potentiometric sensor based on the MIP with the common monomer (i.e., methacrylic acid). This allyl-ß-CD MIP-based potentiometric sensor shows a detection limit of 0.29 µM for BPA, which is about one order of magnitude lower than that obtained by the conventional MIP-based potentiometric sensor. We believe that utilizing a functional monomer with specific recognition ability towards target in the fabrication of MIP could provide an appealing way to construct highly selective MIP-based electrochemical and optical sensors.

Polymeric membrane potentiometric antibiotic sensors using computer-aided screening of supramolecular macrocyclic carriers.

Carrier-based polymeric membrane potentiometric sensors are an ideal tool for detecting ionic species. However, in the fabrication of these sensors, the screening of carriers still relies on empirical trial- and error-based optimization, which requires tedious and time-consuming experimental verification. In this work, computer-aided screening of carriers is applied in the preparation of polymeric membrane potentiometric sensors. Molecular docking is used to study the host-guest interactions between receptors and targets. Binding energies are employed as the standard to screen the appropriate carrier. As a proof-of-concept experiment, the antibiotic ciprofloxacin is selected as the target model. A series of supramolecular macrocyclic receptors including cyclodextrins, cucurbiturils and calixarenes are chosen as potential receptors. The proposed sensor based on the receptor calix[4]arene screened by molecular docking shows a lower detection limit of 0.5 µmol L-1 for ciprofloxacin. It can be expected that the proposed computer-aided screening technique of carriers can provide a simple but highly efficient method for the fabrication of carrier-based electrochemical and optical sensors.

Anti-fouling TiO2-Coated Polymeric Membrane Ion-Selective Electrodes with Photocatalytic Self-Cleaning Properties.

Nowadays, using a polymeric membrane ion-selective electrode (ISE) to achieve reliable ion sensing in complex samples remains challenging because of electrode fouling. To address this challenge, we describe a polymeric membrane ISE with excellent anti-fouling and self-cleaning properties based on surface covalent modification of an anatase TiO2 coating. Under ultraviolet illumination, the reactive oxygen species produced by photocatalytic TiO2 can not only kill microorganisms but also degrade organic foulants into carbon dioxide and water, and a formed superhydrophilic film can effectively prevent the adsorption of foulants, thus inhibiting the occurrence of biofouling and organic fouling of the sensors. More importantly, residual foulants could be fully self-cleaned through the flow of water droplets. By using Ca2+-ISE as a model, an anti-fouling polymeric membrane potentiometric sensor has been developed. Compared to the unmodified electrode, the TiO2-coated Ca2+-ISE exhibits remarkably improved anti-biofouling properties with a low bacterial adhesion rate of 4.74% and a high inhibition rate of 96.62%. In addition, the proposed electrode displays unique properties of anti-organic dye fouling and a superior self-cleaning ability even after soaking in a concentrated bacterial suspension of 109 CFU mL-1 for 60 days. The present approach can be extended to improve the fouling resistance of other electrochemical or optical membrane sensors and is promising for the construction of contamination-free sensors.

Potentiometric sensor based on a computationally designed molecularly imprinted receptor.

Molecularly imprinted polymer (MIP)-based polymeric membrane potentiometric sensors are ideal candidates for detection of organic species. The development of such sensors has opened new attractive horizons for potentiometric sensing. However, it should be noted that in the preparation of these MIP receptors, the selection of the functional monomer usually depends on empirical trial- and error-based optimization, which involves tedious and time-consuming experiments. In this work, the computer-aided design and synthesis of an MIP receptor are applied in the fabrication of an MIP-based potentiometric sensor. The density functional theory calculation with the B3LYP model and 6-31G(d) basis set is used to study the interactions between the functional monomer and template molecules. The binding energies of the complexations between the template molecule and different functional monomers are used as a criterion for the selection of the proper monomer. The designed MIP is then synthesized and employed as the receptor for the fabrication of the potentiometric sensor. As a proof-of-concept experiment, the antibiotic sulfadiazine has been selected as a model and 4 functional monomers, 2-hydroxyethyl methacrylate, methyl methacrylate, N-isopropylacrylamide and N-phenylacrylamide, have been chosen. The designed MIP-based sensor exhibits excellent sensitivity with a linear range of 1-10 µM and also shows a good selectivity. We believe that the proposed computer-aided synthesis technique for the MIP receptor selection can provide a general and facile way to replace the traditional empirical MIP preparation method in the fabrication of MIP-based electrochemical and optical sensors.

Thin membrane-based potentiometric sensors for sensitive detection of polyions.

A novel protocol for development of sensitive and rapid polymeric membrane polyion sensitive electrodes has been explored in this work. In contrast to the traditional polyion electrodes which usually have a sensing membrane thickness of 100∼200 µm, a thin membrane electrode with a membrane thickness of 5 µm is proposed to detect polyions. By using such thin membrane configuration, the diffusion of polyions from the organic boundary layer into the bulk of the membrane can be effectively blocked. The induced accumulation of polyions in the membrane boundary layer largely enhances the obtained potential response. It has been found that the proposed electrode shows a remarkably improved sensitivity and measurement time over conventional potentiometric polyion sensors based on the thick membranes. By using protamine as a model of polyions, the new concept offers a detection limit nearly two orders of magnitude lower than those obtained by the traditional thick-membrane polyion electrodes for potentiometric measurements of polyions. The proposed polyion sensing platform offers great promise in the sensitive and rapid detection of polyions as well as other polyion-involved bioanalyses.

Multifunctional Molecularly Imprinted Receptor-Based Polymeric Membrane Potentiometric Sensor for Sensitive Detection of Bisphenol A.

Molecularly imprinted polymer (MIP)-based polymeric membrane potentiometric sensors have become an attractive tool for detection of organic species. However, the MIP receptors in potentiometric sensors developed so far are usually prepared by only using single functional monomers. This may lead to low affinities of the MIP receptors due to the lack of diversity of the functional groups, thus resulting in low detection sensitivity of the potentiometric sensors. Additionally, these classical MIP receptors are nonconductive polymers, which are undesirable for the fabrication of an electrochemical sensor. Herein, we describe a novel multifunctional MIP receptor-based potentiometric sensor. The multifunctional MIP receptor is prepared by using two functional monomers, methacrylic acid, and 3-vinylaniline with a dual functionality of both recognition and conduction properties. The poly(aniline) groups are introduced into the methacrylic acid-based MIP by postoxidation of the aniline monomer. Such poly(aniline) groups not only serve as the additional functional groups for selective recognition, but also work as a conducting polymer. The obtained multifunctional MIP receptor shows a high binding capacity and an excellent electron-transfer ability. By using bisphenol A as a model, the proposed multifunctional MIP sensor exhibits a largely improved sensitivity and low noise levels compared to the conventional MIP sensor. We believe that the proposed MIP-based sensing strategy provides a general and facile way to fabricate sensitive and selective MIP-based electrochemical sensors.

Towards potentiometric detection in nonaqueous media: Evaluation of the impacts of organic solvents on polymeric membrane ion-selective electrodes.

Polymeric membrane ion-selective electrodes (ISEs) have been widely used in various fields including clinical diagnosis, environmental monitoring and industrial analysis. Although most samples of analytical interest measured by the ISEs are aqueous solutions, the applications of these electrodes in nonaqueous media are often inevitable. Unfortunately, so far, little has been known about the extent to which the properties of the ISEs could be affected by the organic solvents. Herein, the feasibility for the applications of the polymeric membrane ISEs in nonaqueous media has been investigated. A polymeric membrane Ca2+-ISE is chosen as a model of potentiometric sensors. Four typical water miscible organic solvents (three protic solvents: ethanol, acetic acid, and methanol, and one aprotic dipolar solvent: acetonitrile) are used as the representative examples. Experiments show that the aprotic solvent acetonitrile has the strongest destructive ability towards the sensing performance of the ISE in terms of Nernstian slope and selectivity coefficient. Moreover, the effect on the sensing performance depends on the kind of the protic solvent, the immersion time and the polarity of the membrane plasticizer. We believe that the obtained results could promote further applications of the polymeric membrane ISEs in the organic solvent-containing samples, which could significantly extend the application scope of the ISEs.

Enhancing the Oil-Fouling Resistance of Polymeric Membrane Ion-Selective Electrodes by Surface Modification of a Zwitterionic Polymer-Based Oleophobic Self-Cleaning Coating.

Due to the frequent oil spill accidents and pollution of industrial oily wastewater, oil fouling has become a great challenge to polymeric membrane ion-selective electrodes (ISEs) for applications in oil-contaminated areas. Herein, a simple approach is proposed to enhance the oil-fouling resistance of polymeric membrane ISEs by surface modification of a zwitterionic polymer-based underwater oleophobic coating. As a proof-of-concept, a classical poly(vinyl chloride) membrane-based calcium ion-selective electrode (Ca2+-ISE) is chosen as a model sensor. The zwitterionic polymer-based coating can be readily modified on the sensor's surface by immersion of the electrode into a mixture solution of dopamine and a zwitterionic acrylate monomer (i.e., sulfobetaine methacrylate, SBMA). The formed poly(SBMA) (PSBMA) coating alters the oleophilic membrane surface to an oleophobic one, which endows the surface with excellent self-cleaning properties without loss of the sensor's analytical performance. Compared to the pristine Ca2+-ISE, the PSBMA-modified Ca2+-ISE exhibits an improved analytical stability when exposed to oil-containing wastewater. The proposed approach can be explored to enhance the oil-fouling resistance of other polymeric membrane-based electrochemical sensors for use in the oil-polluted environment.

A molecularly imprinted polymer-based potentiometric sensor based on covalent recognition for the determination of dopamine.

Polymeric membrane potentiometric sensors based on molecularly imprinted polymers (MIPs) have been successfully designed for the detection of organic compounds both in ionic and neutral forms. However, most of these sensors are based on the non-covalent recognition interactions between the functional groups of the MIP in the polymeric sensing membrane and the target. These weak non-covalent interactions are unfavorable for the detection of hydrophilic organic compounds (e.g., dopamine). Herein novel MIP potentiometric sensor based covalent recognition for the determination of protonated dopamine is described. Uniform-sized boronate-based MIP beads are utilized as the recognition receptors. These receptors can covalently bind with dopamine with a cis-diol group to form a five-membered cyclic ester and thus provide a higher affinity because of the stronger nature of the covalent bonds. It has been found that the proposed electrode shows an excellent sensitivity towards dopamine with a detection limit of 2.1 µM, which could satisfy the needs for in vivo analysis of dopamine in the brain of living animals. We believe that the covalent recognition MIP-based sensing strategy provides an appealing way to design MIP-based electrochemical and optical sensors with excellent sensing properties.

Thin polymeric membrane ion-selective electrodes for trace-level potentiometric detection.

In this work, we describe a novel method to improve the detection limits of the non-classical polymeric membrane ion-selective electrodes (ISEs) which are conditioned with highly discriminated ions instead of primary ions. It is based on a thin-layer ISE membrane with a thickness of 5 µm, which is coated on ordered mesoporous carbon used as solid contact. The diffusion of the primary ion from the surface of the sensing membrane to the bulk of the membrane could be avoided by the proposed thin membrane configuration. Since the detection sensitivity of the non-classical ISEs depends on the accumulation of the primary ion in the interfacial layer of the sensing membrane, a lower detection limit can be obtained. By using the copper ion as a model, the present potentiometric sensor shows a significantly improved detection sensitivity compared to the conventional ISE with a membrane thickness of ca. 200 µm. Low detection limits of 0.29 and 0.53 nM can be obtained in 0.01 and 0.5 M NaCl, respectively. In addition, the proposed sensor exhibits an excellent reversibility by using a neutral proton-selective ionophore incorporated in the thin membrane.

Plasticizer-free polymer membrane potentiometric sensors based on molecularly imprinted polymers for determination of neutral phenols.

Polymeric membrane potentiometric sensors based on molecularly imprinted polymers (MIPs) as the receptors have been successfully developed for detection of organic and biological species. However, it should be noted that all of the polymeric membrane matrices of these sensors developed so far are the plasticized poly(vinyl chloride) (PVC) membranes, which are usually suffered from undesired plasticizer leaching. Hence, for the first time, we describe a novel plasticizer-free MIP-based potentiometric sensor. A new copolymer, methyl methacrylate and 2-ethylhexyl acrylate (MMA-2-EHA), is synthesized and used as the sensing membrane matrix. By using neutral bisphenol A (BPA) as a model, the proposed plasticizer-free MIP sensor shows an excellent sensitivity and a good selectivity with a detection limit of 32 nM. Additionally, the proposed MMA-2-EHA-based MIP membrane exhibits lower cytotoxicity, higher hydrophobicity and better MIP dispersion ability compared to the classical plasticized PVC-based MIP sensing membrane. We believed that the new copolymer membrane-based MIP sensor can provide an appealing substitute for the traditional PVC membrane sensor in the development of polymeric membrane-based electrochemical and optical MIP sensors.

Stimulus-Responsive Imprinted Polymer-Based Potentiometric Sensor for Reversible Detection of Neutral Phenols.

Nowadays, polymeric membrane potentiometric sensors based on the molecularly imprinted polymers (MIPs) have been successfully developed for detection of various organic and biological species. However, it is difficult for these sensors to perform reversible detection of the targets due to the high affinities of the MIPs toward the targets. In this work, we propose a novel method for fully reversible potentiometric detection of neutral phenols based on the stimulus-responsive MIP as the selective receptor. Since such smart receptor can switch its recognition abilities according to the external environmental stimuli, the MIP binding sites in the polymeric membrane can be regenerated via the stimulus after each measurement. Thus, potentiometric reversible detection of the target can be achieved. As a proof of concept, the pH-responsive MIP is used as the selective receptor, which can be synthesized by using 4-vinylphenylboronic acid as the functional monomer. The boronate-affinity MIP can covalently bind with a cis-diol containing compound to form a five- or six-membered cyclic ester in a weakly alkaline aqueous solution, while the produced ester dissociates when the surrounding pH is changed to acidic. By using catechol as a model, the proposed smart sensor exhibits a significantly improved reversibility compared to the conventional MIP-based sensor. We believed that the stimulus-responsive MIP-based sensing strategy could provide an appealing way to design reversible MIP-based electrochemical and optical sensors.

Improving the Biocompatibility of Polymeric Membrane Potentiometric Ion Sensors by Using a Mussel-Inspired Polydopamine Coating.

Polymeric membrane potentiometric ion sensors have been widely used in clinical diagnosis for the detection of electrolyte ions and account for billions of measurements every year throughout the world. However, in many cases of practical relevance, biofouling, which might lead to sensor failure, usually occurs due to the lack of biocompatibility of these sensors. Herein, we describe a simple and robust approach for improving the biocompatibility of the polymeric ion-selective membranes. A marine mussel-inspired polydopamine polymer is used as a hydrophilic coating on the surface of conventional potentiometric ion sensors. Such a coating can be easily formed by self-polymerization of dopamine and robustly deposited on the sensor surface mimicking the adhesion mechanism of mussels. The classical poly(vinyl chloride) membrane-based calcium ion-selective electrode (ISE) is chosen as a model. Compared to the unmodified Ca2+ ISE, the polydopamine modified electrode shows a significantly reduced blood platelet adsorption while retaining original potentiometric ion response properties, which clearly indicates a high antifouling capability induced by the hydrophilic polydopamine coating. We believe that the proposed approach can provide an appealing way to improve the biocompatibility in the development of polymeric membrane electrochemical and optical sensors.

Robust fabrication of nanomaterial-based all-solid-state ion-selective electrodes.

Currently, nanomaterial-based all-solid-state ion-selective electrodes (ASS-ISEs) have become attractive tools for ion sensing in environmental and biological applications. However, nanomaterial solid contact can easily fall off the electrode surface owing to poor adhesion. This poses serious limits to the wide use of these sensors. Herein, we report a general and facile method for the robust fabrication of nanomaterial-based ASS-ISEs. It is based on the silver-based conductive adhesive (CA) with excellent electronic conductivity and strong adhesion ability as the binder to construct nanomaterial-based solid contact. The solid-contact Ca2+-ISE based on single-walled carbon nanotubes (SWCNTs) is chosen as a model. The proposed electrode based on CA-SWCNTs shows a linear response in the concentration range of 10-6 to 10-3 M with a slope of 25.96 ± 0.36 mV per decade and a detection limit of 1.7 × 10-7 M. In addition, the CA-SWCNT-based Ca2+-ISE exhibits an improved potential stability and reduced water film compared to the coated-wire ISE. Above all, experiments also show that the CA-SWCNT-based electrode exhibits nearly the same electrochemical characteristics as the classical only SWCNT-based electrode in term of resistance, capacitance and potential stability. We believe that CA-nanomaterial-based solid contacts provide an appealing substitute for traditional solid contacts based on nanomaterials.

Soluble Molecularly Imprinted Nanorods for Homogeneous Molecular Recognition.

Nowadays, it is still difficult for molecularly imprinted polymers (MIPs) to achieve homogeneous recognition since they cannot be easily dissolved in organic or aqueous phase. To address this issue, soluble molecularly imprinted nanorods have been synthesized by using soluble polyaniline doped with a functionalized organic protonic acid as the polymer matrix. By employing 1-naphthoic acid as a model, the proposed imprinted nanorods exhibit an excellent solubility and good homogeneous recognition ability. The imprinting factor for the soluble imprinted nanoroads is 6.8. The equilibrium dissociation constant and the apparent maximum number of the proposed imprinted nanorods are 248.5 µM and 22.1 µmol/g, respectively. We believe that such imprinted nanorods may provide an appealing substitute for natural receptors in homogeneous recognition related fields.

Oxidative removal of quinclorac by permanganate through a rate-limiting [3 + 2] cycloaddition reaction.

Quinclorac, a widely used herbicide in agriculture, has been recognized as an emerging environmental pollutant owing to its long persistence and potential risk to humans. However, no related information is available on the degradation of quinclorac by employing oxidants. Herein, the reactivity of quinclorac with permanganate was systematically investigated in water by combining experimental and computational approaches. The reaction followed overall second-order kinetics pointing to a bimolecular rate-limiting step. The second-order rate constant was found to be 3.47 × 10-3 M-1 s-1 at 25 °C, which was independent of pH over the range from 5 to 9 and was dependent on temperature over the range from 19 to 35 °C. The initial product was identified by UPLC-Q-TOF-MS to be mono-hydroxylated quinclorac, which was more susceptible to further oxidation. The result could be supported by the complete simulation of the reaction process in DFT calculations, indicating the [3 + 2] cycloaddition oxidation of the benzene ring in the rate-limiting step. The plausible mechanism was then proposed, accompanied by the analysis of the HOMO indicating the hydroxylation position and of the ESP suggesting a more electron-rich moiety. Considering the high effectiveness and low toxicity, permanganate oxidation was considered to be a very promising technique for removing quinclorac from aquatic environments.

PI3K signaling pathways modulated white spot syndrome virus (WSSV) replication in Procambarus clarkii.

The PI3K/AKT signaling pathway is commonly exploited to regulate viral replication and affect the fate of infected cells. In the present study, a PI3K-specific inhibitor (LY294002) was employed to pretreat crayfish to evaluate the effects of PI3K/AKT signaling pathway in WSSV replication. The results showed that the WSSV copy numbers in crayfish pretreated with LY294002 were significantly lower than those in Tris-HCl pretreatment crayfish on the sixth and tenth day after WSSV infection. In semigranular cells, the apoptosis rates were up-regulated on the third day post-WSSV infection, and a significantly lower proportion of apoptosis cells were observed in LY294002-pretreatment group. The expression level of Bax, Bax inhibitor-1 and lectin mRNA in haemocytes of crayfish were increased after WSSV infection. After the secondary stimulation with Tris-HCl, the Bax expression level in LY294002-pretreatment crayfish was significantly higher than that of crayfish pretreated with Tris-HCl on the third or sixth day, but the Toll and lectin mRNA expression decreased significantly on the third, sixth and tenth day. The Bax mRNA expression levels in LY294002-WSSV group were significantly higher than those in Tris-HCl-WSSV group on the third and tenth day. The Bax inhibitor-1 mRNA expression levels in LY294002-WSSV group were significantly lower than those in Tris-HCl-WSSV crayfish on the third day. These results together indicated that the hosts PI3K/AKT signaling pathway play positive roles in WSSV replication through the balance between host cell apoptois and innate immune responses. This information is helpful to further understand the role of PI3K/AKT signaling pathway on WSSV replication in Decapoda crustaceans.