SENSORS AND ACTUATORS B-CHEMICAL
·
2024
·
IF 8
Luting Yan,Yuhao Zhang,Qi Guo,Lei Tian,Falan Li,Qingqing Yang,Yanyan Zhang
Most electrochemiluminescent (ECL) biosensors currently necessitate coreactants that affect sensor detection accuracy and increase sensor construction complexity. This study detailed the creation of an ECL biosensor without coreactant, utilizing Methionine-capped Au nanoclusters (Met-AuNCs) as luminophore and a Pb 2+ -dependent DNAzyme-mediated DNA walker signal amplification strategy to detect Vibrio parahaemolyticus (Vp) in scallops. Double-stranded DNA (dsDNA) was produced through hybridization of the Vp aptamer with DNA walker. The electrode surface was loaded with cDNA labeled with Met-AuNCs. Upon the presence of Vp, it competitively bound to the aptamer, resulting in dsDNA dissociation and exposing the DNAzyme sequence in the DNA walker. The introduction of Pb 2+ initiated the activation and circulation of the DNAzyme-mediated DNA walker on the electrode surface, causing the cDNA to be sheared. As a result, the cDNA-Met-AuNCs on the surface of the electrode decreased, resulting in a reduction of ECL signal intensity. The ECL biosensor detected Vp at a range of 1.00×10 1 -1.00×10 7 CFU/mL, under optimal conditions, with the limit of detection (LOD) being 1.23 CFU/mL. In this work, the biosensor demonstrated excellent stability and specificity and had proven successful in detecting actual samples of aquatic products. This study offers a novel ECL method without a coreactant for the detection of foodborne pathogens in aquatic products.
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MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Wei Zhang,Qiao Tang,Mengmeng Wang,Pei Liang,Xuejiao Zhang,Kun Pang,Qiang Chen,Xiaoping Yu
In this work, an aptamer sensor (aptasensor) for SERS detection of Vibrio parahaemolyticus was prepared by utilizing flower-shaped silver nanoparticles coated with graphene oxide (Ag@GO) and core–shell Au@Ag nanoparticles embedded with the Raman reporter 4-mercaptobenzoic acid (4MBA). Graphene oxide (GO) was synthesized using an improved Hummers method and wrapped around thiol-functionalized Ag nanoflowers to form the Ag@GO composite. The SERS aptamer sensor with the strongest Raman signal of 4MBA was developed by combining Ag@GO with Au@4MBA@Ag NPs modified by aptamer through hydrogen bonding and π-π stacking interactions. The aptasensor exhibited good repeatability and stability, and demonstrated high specificity. And it showed a good linear relationship for Vibrio parahaemolyticus detection within the concentration range of 10 2 cfu/mL–10 7 cfu/mL. Introducing GO as an internal standard improved the linear regression coefficient (R 2 ) of detection from 0.9885 to 0.9956. Additionally, spiked real samples of fresh shrimp were tested, yielding recovery rates between 94.1 % and 104.7 %, which demonstrate the feasibility and practicality of this method.
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FOOD CONTROL
·
2024
·
IF 5.6
Hongyong Lin,Tengyang Liao,Yaoyao Sang,Jin Mao,Yu Liu,Bin Zhang,Di Sun,Wei Jiang
Highly accurate and sensitive detection of Vibrio parahaemolyticus is crucial for ensuring seafood safety and preventing the transmission of foodborne diseases. Herein, a dual-recognition strategy-driven colorimetric biosensor for the detection of V. parahaemolyticus was developed. This biosensor was based on the use of PEG-mediated cefepime-functionalized magnetic nanoparticles (Cefe-PEG-MNPs) and aptamer-modified Fe-doped polydopamine (Fe@PDA) nanozymes. The magnetic platform Cefe-PEG-MNPs was introduced to enhance the enrichment efficiency of V. parahaemolyticus , achieving a capture efficiency exceeding 90%. The Fe@PDA nanozyme exhibited notable peroxidase mimetic enzyme activity. The colorimetric biosensor developed for V. parahaemolyticus detection demonstrated good sensitivity, with a detection range spanning from 2.1 × 10 1 to 2.1 × 10 6 CFU/mL, and a limit of detection of 21 CFU/mL. Moreover, the biosensor demonstrated remarkable specificity, achieving good recovery rates from 95.80% to 103.80% in Penaeus vannamei with a relative standard deviation of less than 1.38%. The newly developed colorimetric biosensor offers a remarkable opportunity for the accurate and sensitive detection of V . parahaemolyticus in seafood.
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Huan Lin,Qiaoling Chen,Danfeng Zhang,Fang Cao,Yonghua Hu,Qiuqiang Zhu,Yining Huang,Tingting Liu,Qiaoping Sun
This study develops a dual-dynamic-crosslinked hydrogel via Schiff base and borate ester networks using dialdehyde cellulose (DAC) and ε-polylysine (ε-PL), with tunable properties and breakthroughs in combating drug resistance. DAC (3.83 ± 0.25 mmol/g aldehyde content, 80.21 % yield) crosslinked with ε-PL at 1:0.45 M ratio yields DAC-ε-PL-3 (92.37 % productivity, 8.26 mg/g lysine). The PVA-DAC-ε-PL hydrogel features 1.05–8.10 μm pores, 85.56–93.19 % water retention, shear-thinning rheology, and excellent mechanical properties. It shows broad-spectrum activity against 9 pathogens, including multidrug-resistant (MDR) Vibrio parahaemolyticus (HG9G, HG3G, HC3G), with an 8.75 mg/mL MIC against HG9G and 12.35–16.97 mm inhibition zones. The antibacterial mechanism against MDR strains involves membrane disruption (membrane disruption observed by scanning electron microscopy, elevated conductivity, AKP leakage), respiratory chain inhibition, and downregulation of toxR , tetA , and sul2 . Sustained activity inhibits MDR strains for 24 h. This eco-friendly platform addresses antibiotic resistance, with promise for wound dressings and food packaging.
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Marine Drugs
·
2022
·Vol. 20
·期 11
·702
·
IF 6.085
Lefan Li,Fengyan Liang,Chengpeng Li,Tingting Hou,Dong-an Xu
To explore the application of chitosan–gentamicin conjugate (CS-GT) in inhibitingVibrio parahaemolyticus(V. parahaemolyticus), which is an important pathogen in aquatic animals worldwide, the antimicrobial activity of CS-GT and the effects of a CS-GT dose on the intestine histopathology and intestinal flora ofV. parahaemolyticus-infected shrimps were explored. The results showed that CS-GT possessed broad-spectrum antibacterial activity, with minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and half inhibitory concentration (IC50) of 20.00 ± 0.01, 75.00 ± 0.02 and 18.72 ± 3.17 μg/mL forV. parahaemolyticus, respectively. Further scanning electron microscope and cell membrane damage analyses displayed that the electrostatic interaction of CS-GT with cell membrane strengthened after CS grafted GT, resulting in leakage of nucleic acid and electrolytes ofV. parahaemolyticus. On the other hand, histopathology investigation indicated that high (100 mg/kg) and medium (50 mg/kg) doses of CS-GT could alleviate the injury of a shrimp’s intestine caused byV. parahaemolyticus. Further 16S rRNA gene sequencing analysis found high and medium dose of CS-GT could effectively inhabitV. parahaemolyticusinvasion and reduce intestinal dysfunction. In conclusion, CS-GT possesses good antibacterial activity and could protect shrimps from pathogenic bacteria infection.Keywords:chitosan;Vibrio parahaemolyticus;antimicrobial mechanism;intestine microbe
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Food Analytical Methods
·
2018
·Vol. 12
·期 2
·595-603
·
IF 2.245
Shen Mofei,Duan Nuo,Wu Shijia,Zou Ying,Wang Zhouping
This paper investigated a new detection method for food pathogenic bacteria Vibrio parahaemolyticus based on surface-enhanced Raman scattering (SERS). Nanostructures consisting of Aptamer-Au nanoparticles-polydimethylsiloxane (Apt-Au-PDMS film) were firstly used as SERS substrate. Aptamers were immobilized on the functionalized PDMS film covered with Au nanoparticles (AuNPs) to act as a capture structure via the affinity binding of aptamer, and Vibrio parahaemolyticus . AuNPs were modified with the aptamer and Raman reporter molecule 4-mercaptobenzoic acid (4-MBA) to act as SERS-sensing probes that bind to the target in the same way as the Apt-Au-PDMS film. The solid sandwich structure was formed by capture structure-target-SERS-sensing probes. The concentration of Vibrio parahaemolyticus can be quantified by measurement of the SERS intensity of 4-MBA. Under optimal conditions, the signal of 4-MBA at 1590 cm −1 was linearly related to the Vibrio parahaemolyticus concentration in the range between 1.2 × 10 2 and 1.2 × 10 6 cfu·mL −1 . Recoveries ranging from 97.7% to 105.3% were found when analyzing spiked prawn samples. This developed ultrasensitive aptamer-based SERS detection structure suggested that it would be a promising strategy for a variety of sensing applications.
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Frontiers in Immunology
·
2026
·
IF 7
Yihuan Chen,Liyuan Yi,Le Zhang,Yiming Zheng,Chen Chen,Yaohua Wang,Shilin Shang,Maocang Yan,Qianjin Zhou,Jiong Chen
Acute hepatopancreatic necrosis disease (AHPND), caused by Vibrio parahaemolyticus, poses a severe threat to global shrimp aquaculture. Reduced susceptibility of aquaculture-associated bacterial pathogens to conventional antibiotics, together with concerns regarding antimicrobial input and environmental exposure, highlights the need for dose-sparing therapeutic strategies. This study investigated the antibiotic-potentiating activity of honokiol (HKL), a plant-derived lignan, in combination with sulfamonomethoxine sodium (SMM-Na) against AHPND. Checkerboard and time-kill assays demonstrated that sub-inhibitory HKL concentrations (32–64 μg/mL) markedly enhanced SMM-Na activity, reducing its minimum inhibitory concentration by 8–32-fold and achieving bactericidal effects in combination. The SMM-Na + HKL combination also significantly inhibited biofilm formation and compromised bacterial membrane integrity, as evidenced by enhanced propidium iodide uptake and scanning electron microscopy, while inducing severe ATP depletion. In Penaeus vannamei challenged with V. parahaemolyticus, dietary administration of low-dose SMM-Na (16 mg/kg) and HKL (32 mg/kg) yielded the highest survival rate (86.6%), substantially reduced hepatopancreatic bacterial load, attenuated histopathological damage, and restored total hemocyte counts. The treatment also upregulated a broad spectrum of immune-related genes (e.g., Alf, Tlr, Lec, Crustin, Lzm, CatB) in the hepatopancreas and mucosal barrier genes (e.g., Muc-1, Muc-4, Muc-5AC, Muc-19) in the intestine. Furthermore, 16S rRNA sequencing indicated that the combination therapy was associated with treatment-related shifts in the intestine microbiota, including increased alpha diversity indices, reduced Vibrio abundance, and altered predicted KEGG functional profiles. Collectively, our results identify HKL as a natural-origin antibiotic potentiator that enhances the efficacy of low-dose SMM-Na against AHPND-causing V. parahaemolyticus. These findings provide proof-of-concept evidence for a dose-sparing combination strategy that may reduce therapeutic SMM-Na input while supporting pathogen control and host recovery in shrimp aquaculture.
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MARINE BIOTECHNOLOGY
·
2026
·Vol. 28
·期 3
·93
·
IF 2.8
Liu Fengyi,Geng Qilin,Sun Xue,Lin Jie,Zhu Xuan,Wang Danyang,Zhang Xiao-Hua,Hu Jingjie,Wang Shi,Bao Lisui,Lv Jia
Vibrio parahaemolyticus ( V. parahaemolyticus ), a significant pathogen in aquatic products, necessitates rapid and precise detection to prevent foodborne disease outbreaks. Early detection of bacterial pathogens is crucial for effective prevention and control. In this study, we developed a field-deployable, “sample-in, answer-out” workflow that integrates efficient DNA extraction with visual fluorescence loop-mediated isothermal amplification (LAMP) for the on-site detection of V. parahaemolyticus . This extraction protocol enables efficient one-pot DNA release within 10 min, eliminating the need for additional purification steps. In addition, coupled with an optimized fluorescence LAMP assay targeting V. parahaemolyticus , the entire process—from sample to result—can be completed within approximately 50 min. The optimized assay exhibited high sensitivity, detecting as low as 1 copy/µL of plasmid DNA and 10 CFU per reaction from pure bacterial culture and spiked seawater samples. This extraction-amplification integrated approach demonstrates robust performance and high practicality for on-site early warning and pathogen monitoring in aquaculture safety settings.
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LWT-FOOD SCIENCE AND TECHNOLOGY
·
2022
·
IF 6.056
Xiangpeng Han,Mengyu Zhang,Jiayi Peng,Jinsong Wu,Qingping Zhong
Vibrio parahaemolyticus and its biofilm threaten food safety and human health. Bacteriocins are known to inhibit various foodborne bacteria and their biofilms; however, few bacteriocins with antibiofilm activity against V. parahaemolyticus have been reported. Herein, a novel plantaricin Z057 from Lactiplantibacillus plantarum Z057 was extracted with ethyl acetate, purified by Sephadex LH-20 gel chromatography and reversed-phase high-performance liquid chromatography, and identified by nano-liquid chromatography coupled with tandem mass spectrometry. Its molecular mass and amino acid composition were 1272.61 Da and A-G-G-D-G-G-H-G-G-S-I-I-L-Y, respectively. Plantaricin Z057 presented a broad antibacterial spectrum against Gram-positive and Gram-negative pathogens. Moreover, it also exhibited excellent heat tolerance, maintained antibacterial activity at pH 2.0–4.0, and was susceptible to common proteases (pepsin, trypsin, papain, proteinase K). In addition, scanning electron microscopy revealed that plantaricin Z057 destroyed membrane integrity and caused cell deformation and cytoplasmic leakage. The crystal violet staining and XTT reduction assays revealed that plantaricin Z057 markedly decreased biofilm biomass and metabolic activity of V. parahemolyticus . Fluorescence microscopy showed that plantaricin Z057 efficiently inactivated sessile cells, and damaged integrated biofilm structure by reducing the extracellular polymer content. In summary, plantaricin Z057 exhibited significant potential for controlling V. parahaemolyticus and could be used in food production as a biological preservative.
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MICROCHEMICAL JOURNAL
·
2021
·
IF 4.821
Yunxia Hou,Liangliang Zhu,Hongshun Hao,Zuowei Zhang,Chao Ding,Gongliang Zhang,Jingran Bi,Shuang Yan,Guishan Liu,Hongman Hou
In this work, layer-by-layer self-assembly method was used to construct the working electrode of the photoelectrochemical (PEC) aptamer sensor. Rare-earth doped Bi 2 WO 6 (BWO-1) and Ag 2 S are combined as highly efficient photoactive material to reduce the recombination rate of electron-hole pairs and enhance the photocurrent responsiveness under visible light. The PEC aptamer sensor, with BWO-1 concentration of 12 mg/mL, AgNO 3 concentration of 0.14 mol/L, ascorbic acid (AA) concentration of 0.13 mol/L, pH value of 7.4, aptamer concentration of 2 μmol/L, and incubation time of 50 min, had an optimal photocurrent responsiveness. The developed method showed a wide linear range from 3.2 × 10 2 CFU/mL to 3.2 × 10 8 CFU/mL. The detection limit was 40 CFU/mL. The constructed PEC aptamer sensor exhibited good detection specificity, stability and reproducibility for Vibrio parahaemolyticus and provided a simple, low-cost and rapid detection strategy for prevention and control of foodborne diseases. This method could be promising for the rapid detection of other food-borne pathogenic bacteria in the food.
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FISH & SHELLFISH IMMUNOLOGY
·
2011
·
IF 3.044
Rui Liu,Jixiang Chen,Kesheng Li,Xiaohua Zhang
A putative serine protease gene was cloned from the genomic DNA of Vibrio parahaemolyticus FYZ8621.4. The gene consisted of 1779 base pairs and encoded a 592 amino acid protein. The gene was expressed in Escherichia coli . The expressed protease was purified by Ni-NTA His-Bind Resin column and showed a 63 kDa band on SDS–PAGE. The protease exhibited proteolytic activity on gelatin agar plate and showed maximal proteolytic activity at pH 8.0 and 37 °C. It hydrolyzed N-α-benzoyl- l -tyrosine p-nitroanilide (BAPNA), but did not N-benzoyl- l -arginine ethylester (BAEE), N-benzoyl- l -tyrosine ethylester (BTEE) and N-acetyl- l -tyrosine ethylester (ATEE). Mutants at conserved residues Asp 51 (Asp 51 –Asn), His 89 (His 89 –Asp) and Ser 318 (Ser 318 –Leu, Ser 318 –Pro) lost proteolytic activities completely. The protein was confirmed to belong to serine protease. The purified serine protease was toxic to zebrafish with a LD 50 of 15.4 μg/fish. A DNA vaccine was constructed by inserting the mutated serine protease (Ser 318 –Pro) gene into pEGFP-N1 plasmid. The pEGFP-N1/m- vps was transfected in HeLa cells. The serine protease was confirmed to be expressed by fluorescence microscopy observation and Western blotting analysis. The pEGFP-N1/m- vps was further observed to express in muscle of the injected turbot ( Scophthalmus maximus ) by Western blotting seven days after immunization. Efficient protection against lethal V. parahaemolyticus challenge was observed on the vaccinated turbot with pEGFP-N1/m- vps , with the highest relative percent survival (RPS) of 96.11%. Significant specific antibody responses were also observed in the turbot vaccinated with the DNA vaccine. The results indicated that the serine protease might be a potential virulence factor and could be used as an efficient vaccine candidate for the disease control caused by V. parahaemolyticus .
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MICROCHIMICA ACTA
·
2024
·Vol. 191
·期 12
·1-12
·
IF 5.3
Chen Yu,Li Ruiwen,Shen Hao,Li Nana,Gao Wenjing,Guo Haipeng,Feng Bin,Yu Shaoning
As one of the most harmful pathogenic bacteria in shrimp aquaculture, Vibrio parahaemolyticus often causes massive mortality in shrimp. Accurate and rapid detection of V. parahaemolyticus in shrimp farming is essential for avoiding huge economic losses caused by related diseases. In this study, we designed a dual-recognition platform for efficient identification and quantification of V. parahaemolyticus . First, the target bacterium was captured with magnetic beads functionalized by aptamers (Apt-MBs), and then, the broad-spectrum fluorescent probe FcMBL@CdSe-ZnS was used to detect the bacterium based on the interactions between fragment crystallizable mannose-binding lectin (FcMBL) and pathogenic bacteria. The proposed dual-recognition strategy centered around aptamers and FcMBL@CdSe-ZnS was applied to definite quantification of V. parahaemolyticus over a wide range of 10–10 8 CFU/mL with a limit of detection of 4 CFU/mL within 55 min. The feasibility was demonstrated by using the platform to detect V. parahaemolyticus from shrimp intestine, aquaculture water, and seawater. Graphical
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