FOOD CONTROL
·
2024
·
IF 5.6
Aiping Deng,Xuan Li,Xiaoxing Fan,Yajie Li,Yelin Han,Tianzhu Guan,Shuo Wang,Guoqiang Zhu,Zhenquan Yang,Wenyuan Zhou
Bacteriophage-based biosensors have the potential for rapid, convenient, and sensitive detection of Staphylococcus aureus in food products; however, their effectiveness is often limited by the host range of the bacteriophage. This study presents a new colorimetric biosensor using the S. aureus phage SapYZU04 exhibiting a broad host range, and a MnFeO nanozyme for the effective detection of S. aureus in food products. Notably, SapYZU04 exhibited excellent features, such as a high titre (1.0 × 10 10 PFU/mL), a short latency period (5 min), a large burst size (210 PFU/cell), a wide host spectrum (98.95%, 94/95), and good stability at pH 3.0–12.0. The biosensor, referred to as SapYZU04@MnFeO was constructed by immobilising SapYZU04 on the MnFeO nanozyme via electrostatic interactions. SapYZU04@MnFeO demonstrated a broad detection range of bacterial concentrations, particularly within the range of 3 × 10 2 to 3 × 10 9 CFU/mL, with a low detection limit of 65 CFU/mL and rapid detection time (14 min). Furthermore, it exhibited high accuracy in detecting and quantifying live S. aureus with recovery rates ranging from 92.21% to 105.56%. The chromogenic system demonstrated high selectivity for S. aureus and was unaffected by other interfering microorganisms or food preservatives, highlighting its specificity and interference-resistance. Whole-genome sequencing of SapYZU04 suggested that the selectivity, sensitivity, and specificity of SapYZU04@MnFeO could be attributed to the tail fibre proteins of the phage. In conclusion, the SapYZU04@MnFeO biosensor is a highly effective tool for identifying viable S. aureus strains in the food industry.
研究领域:
微生物学
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食品安全
·
生物传感技术
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PLoS Pathogens
·
2026
·Vol. 22
·期 2
·e1013975
·
IF 4.9
Huijie Hu,Naiyuan Jiang,Juxiong Liu,Junlong Bi,Xuanting Liu,Bin Xu,Yu Cao,Wenjin Guo,Shoupeng Fu
Staphylococcus aureus ( S. aureus )–driven senescence of bovine mammary epithelial cells is a key determinant of mammary gland health, yet its molecular basis remains poorly defined. Sirtuin 5 (SIRT5), a mitochondria-localized desuccinylase, may play an important regulatory role in this process. This study aimed to elucidate the mechanisms by which S. aureus drives cellular senescence and to define the contribution of the SIRT5–mitochondrial axis to delaying senescence. We found pronounced oxidative stress and cellular senescence in mammary tissues from cows with S. aureus mastitis, accompanied by marked downregulation of SIRT5. In an S. aureus -infected epithelial cell model, infection induced mitochondrial stress characterized by excessive mitochondrial fragmentation, loss of membrane potential, and increased mitochondrial superoxide, along with oxidative damage and cellular senescence. Mechanistically, S. aureus toxins and the toxin-induced inflammatory response cooperatively drove mitochondrial stress, which in turn increased intracellular bacterial burden and exacerbated cell death. During infection, SIRT5 protein abundance was significantly reduced. Mass spectrometry and co-immunoprecipitation analyses indicated that infection upregulated the ubiquitin-conjugating enzyme ubiquitin C (UBC), enhanced its interaction with SIRT5, and promoted ubiquitin-mediated degradation of SIRT5. Loss of SIRT5 increased succinylation of dynamin-related protein 1 (DRP1), inhibited its ubiquitin-mediated degradation, and led to its excessive accumulation on the outer mitochondrial membrane, thereby promoting excessive mitochondrial fission. Functionally, SIRT5 overexpression markedly alleviated mitochondrial stress, oxidative damage, and senescence phenotypes. When mitochondrial fission was forcibly enhanced, the cytoprotective effect of SIRT5 was substantially weakened, confirming that SIRT5 acts through a pathway dependent on mitochondrial integrity. Collectively, S. aureus infection releases toxins and induces inflammatory injury, during which UBC-mediated SIRT5 degradation activates DRP1-dependent mitochondrial hyper-fragmentation, aggravating mitochondrial stress, oxidative stress, and mammary epithelial cell senescence. These findings identify SIRT5 as a critical regulator of redox and mitochondrial homeostasis in mammary epithelial cells and a potential therapeutic target for mitigating oxidative damage associated with bovine mastitis.
Staphylococcus aureus ( S. aureus ) is a leading cause of bovine mastitis, an intramammary infection that reduces milk yield, increases antibiotic use, and results in substantial economic loss. One way this bacterium damages the udder is by driving bovine mammary epithelial cells into premature senescence, but how infection triggers this process has not been fully understood. Here, we show that S. aureus infection induces an inflammatory response via its secreted toxins, which in turn provokes mitochondrial stress. We identify the mitochondrial enzyme Sirtuin 5 (SIRT5) as a key defender in this context. During infection, the host ubiquitin-conjugating enzyme ubiquitin C (UBC) targets SIRT5 for degradation. Loss of SIRT5 leads to accumulation of the mitochondrial fission protein dynamin-related protein 1 (DRP1), causing excessive mitochondrial fragmentation, enhanced oxidative damage, increased intracellular bacterial burden, and aggravated cellular senescence. Restoring SIRT5 helps to preserve mitochondrial integrity and reduces oxidative injury and senescence; however, this protection is weakened when mitochondrial fission is forcibly maintained at a high level. Together, our findings reveal that the UBC–SIRT5–DRP1 pathway links S. aureus infection to mitochondrial damage and cellular senescence, and they highlight SIRT5 as a potential therapeutic target to protect the bovine mammary gland (S1 Fig).
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SENSORS AND ACTUATORS B-CHEMICAL
·
2015
·
IF 4.097
Ting Wang,Zhang Zhang,Yiyan Li,Guoming Xie
Rapid detection of drug-resistant bacteria can facilitate the early diagnosis of various infections and provide essential guidance for appropriate usage of antibiotics. An ultrasensitive electrochemical DNA (E-DNA) sensor based on isothermal strand-displacement polymerization reaction (ISDPR) for detection of mecA gene in methicillin-resistant Staphylococcus aureus (MRSA) was proposed in this study. Methylene blue (MB)-labeled hairpin probes were self-assembled on a gold electrode to confine MB molecules close to the electrode surface for efficient electron transfer. The hairpin probes were then hybridized with complementary target DNA and undergone conformational changes, which led to a decreased electrochemical response. The primers annealed with the opened stems of hairpin probes were extended by DNA polymerase, which in turn released the target DNA to trigger the next polymerization cycle. Finally, each target DNA underwent through many rounds, resulting in a mass of MB molecules moved away from the electrode surface, which generated significant amplified current suppression for mecA gene detection. With this target recycling amplification strategy, the proposed DNA biosensor could offer excellent analytical performance for the detection of mecA gene and provide a new electrochemical method for the early diagnosis of drug-resistant bacteria.
研究领域:
微生物学
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分子生物学
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生物医学工程
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ANALYTICAL CHEMISTRY
·
2014
·
IF 5.825
Lihong Li,Zhao Li,Wen Shi,Xiaohua Li,Huimin Ma
A highly sensitive and selective near-infrared (NIR) fluorescent probe, (E)-2-(2-(6-((2-carboxy-8-oxo-7-(2-phenylacetamido)-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methoxy)-2,3-dihydro-1H-xanthen-4-yl)vinyl)-3,3-dimethyl-1-propyl-3H-indol-1-ium (1), is developed for the determination of β-lactamase. The probe is designed and synthesized by incorporating the specific substrate (cephalosporin) of β-lactamase into a stable hemicyanine skeleton. The fluorescence of 1 itself is very weak due to the alkylation of the hydroxyl group of the hemicyanine fluorophore; however, β-lactamase can selectively react with its substrate (β-lactam ring) in the probe, thereby causing a spontaneous fragmentation. This action leads to the release of the fluorophore and a large fluorescence enhancement at 707 nm (λex = 680 nm). Under the optimized conditions, the fluorescence response of probe 1 is directly proportional to the concentration of β-lactamase in the range of 0.05–2 nM, with a detection limit of 0.02 nM. The validity of the probe has been confirmed by determining β-lactamase in human urine samples in comparison with that determined by iodimetry. Moreover, by taking advantage of its high sensitivity and NIR emission feature, the probe has also been utilized to image β-lactamase in three types of Staphylococcus aureus, including methicillin-resistant S. aureus ATCC BAA44, penicillin-resistant strain ATCC 11632, and penicillin-susceptible strain ATCC 29213, which clearly reveals the significantly different expression levels of β-lactamase in these S. aureus.
研究领域:
微生物学
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生物医学工程
·
药物化学
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高影响因子
Foodborne Pathogens and Disease
·
2023
·Vol. 20
·期 3
·90-99
·
IF 2.8
Wenyi Ran,Yuxi Yue,Feiwu Long,Kai Zhong,Jinrong Bai,Yue Xiao,Qian Bu,Yina Huang,Yanping Wu,Hong Gao
Staphylococcus aureus is a major foodborne pathogen that leads to various diseases due to its biofilm and virulence factors. This study aimed to investigate the inhibitory effect of 2R,3R-dihydromyricetin (DMY), a natural flavonoid compound, on the biofilm formation and virulence of S. aureus, and to explore the mode of action using transcriptomic and proteomic analyses. Microscopic observation revealed that DMY could remarkably inhibit the biofilm formation by S. aureus, leading to a collapse on the biofilm architecture and a decrease in viability of biofilm cell. Moreover, the hemolytic activity of S. aureus was reduced to 32.7% after treatment with subinhibitory concentration of DMY (p < 0.01). Bioinformation analysis based on RNA-sequencing and proteomic profiling revealed that DMY induced 262 differentially expressed genes and 669 differentially expressed proteins (p < 0.05). Many downregulated genes and proteins related to surface proteins were involved in biofilm formation, including clumping factor A (ClfA), iron-regulated surface determinants (IsdA, IsdB, and IsdC), fibrinogen-binding proteins (FnbA, FnbB), and serine protease. Meanwhile, DMY regulated a wide range of genes and proteins enriched in bacterial pathogenesis, cell envelope, amino acid metabolism, purine and pyrimidine metabolism, and pyruvate metabolism. These findings suggest that DMY targets S. aureus through multifarious mechanisms, and especially prompt that interference of surface proteins in cell envelope would lead to attenuation of biofilm and virulence.
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INDUSTRIAL CROPS AND PRODUCTS
·
2022
·
IF 6.449
Meiling Sun,Zhenghui Luo,Qi Yang,Chunxu Huang,Yu Kuang,Mei Kang,Jie Liu
Methicillin-resistant Staphylococcus aureus (MRSA) has evolved to tolerate almost all known β-lactam antibiotics, and been exhibiting resistance to vancomycin . Developing new drugs against the MRSA infection are urgently needed. Natural resource is enriched with antimicrobial molecules. Here, we report an extract from leaves of Cyclocarya paliurus (ECPL), a species of tree native to China and being traditionally cultivated as medicinal plant , which shows a broad spectrum of bacteriostatic and bactericidal effects at MIC 50 15.99 ± 0.55 μg/ml against methicillin-susceptible and -resistant S. aureus . The ECPL suppresses bacterial expression of FtsZ at both transcriptional and protein levels, inhibits its GTPase activity, blocks the assembly dynamics of FtsZ, and disrupts the formation of Z-ring. The bacteria cells fail to divide, enlarge initially, and lyse eventually. In mouse model, the ECPL facilitates skin wound healing with MRSA infection as effective as vancomycin. Our study unveils the potential of identifying novel molecule in ECPL for development of new class of antibiotic against both methicillin-susceptible and -resistant S. aureus , and support domestication of the C. paliurus growing from wild to cultivated, and further scale up the cultivation into industrialized farm for sustainable harvest for pharmaceutic interest.
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MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 9
·1-9
·
IF 5.479
Pang Bo,Zheng Yan,Wang Juan,Liu Yushen,Song Xiuling,Li Jinhua,Yao Shuo,Fu Kaiyue,Xu Kun,Zhao Chao,Li Juan
A colorimetric test is described for the rapid detection of Staphylococcus aureus (SA). Gold nanorods (AuNRs) are first labeled with urease and yolk immunoglobulin (IgY). This probe can specifically bind SA. In the next step, nonspecific magnetic beads and sample are added. This leads to the formation of the AuNR-IgY-SA-nMB immunocomplex which is then magnetically separated. Finally, a solution of urea is added to the supernatant. Ureases catalyzes the decomposition of urea which results in an increase in the pH value. The increase in the pH value is detected by using a phenolphthalein test paper which undergoes a color change from white to pink. The analytical process can be completed within 20 min. The method is highly specific and can detect as little as 476 cfu·mL −1 . It was verified by analyzing contaminated Chinese cabbage and beef samples, and 1000 cfu·mL −1 of SA were accurately detected.
Schematic representation of a colorimetric method for the detection of Staphylococcus aureus based on the immunocomplex formed from dual-labeled gold nanorod (AuNR) probe, bacteria and non-specific magnetic bead (nMB). This method can be completed within 20 min.
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TALANTA
·
2022
·
IF 6.556
Wenjing Gao,Bin Li,Ling Ling,Li Zhang,Shaoning Yu
Differentiating methicillin-sensitive and methicillin-resistant Staphylococcus aureus (MRSA and MSSA) is crucial for clinical diagnosis and anti-microbial treatment. Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) is an efficient tool for identifying pathogenic microorganisms at the bacterial species level. Here, we found that MRSA and MSSA can be differentiated by MALDI-TOF MS by employing (E)-propylα-cyano-4-hydroxyl cinnamylate (CHCA-C3) as the matrix, which shows great performance for proteins/peptides, especially hydrophobic proteins. The results show that the mass spectra profile of standard MRSA (ATCC 43300) is significantly different from the profiles of standard MSSA strains (ATCC 25923 and 29213) when using CHCA-C3 as the matrix compared to traditional matrix. The mass profiles had great reproducibility and were scarcely influenced by the growth medium. Due to the enhanced discrimination ability of CHCA-C3, we collected the mass spectra of 62 clinical S. aureus strains and selected four representative peaks for principal component analysis, which showed great differentiation. Our results suggest that employing a suitable matrix could enhance the discrimination ability of antibiotic-resistant bacteria by MALDI-TOF MS.
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Nanomaterials
·
2022
·Vol. 12
·期 11
·1865
·
IF 5.719
Weiwei Zhang,Zhao Kuang,Ping Song,Wanzhen Li,Lin Gui,Chuchu Tang,Yugui Tao,Fei Ge,Longbao Zhu
Photothermal therapy has been widely used in the treatment of bacterial infections. However, the short photothermal effective radius of conventional nano-photothermal agents makes it difficult to achieve effective photothermal antibacterial activity. Therefore, improving composite targeting can significantly inhibit bacterial growth. We inhibited the growth ofStaphylococcus aureus(S. aureus) by using an extremely low concentration of vancomycin (Van) and applied photothermal therapy with molybdenum disulfide (MoS2). This simple method used chitosan (CS) to synthesize fluorescein 5(6)-isothiocyanate (FITC)-labeled and Van-loaded MoS2-nanosheet hydrogels (MoS2-Van-FITC@CS). After modifying the surface, an extremely low concentration of Van could inhibit bacterial growth by trapping bacteria synergistically with the photothermal effects of MoS2, while FITC labeled bacteria and chitosan hydrogels promoted wound healing. The results showed that MoS2-Van-FITC@CS nanosheets had a thickness of approximately 30 nm, indicating the successful synthesis of the nanosheets. The vitro antibacterial results showed that MoS2-Van-FITC with near-infrared irradiation significantly inhibitedS. aureusgrowth, reaching an inhibition rate of 94.5% at nanoparticle concentrations of up to 100 µg/mL. Furthermore, MoS2-Van-FITC@CS could exert a healing effect on wounds in mice. Our results demonstrate that MoS2-Van-FITC@CS is biocompatible and can be used as a wound-healing agent.Keywords:MoS2nanosheet;antibacterial activity;photothermal therapy;wound-healing
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JOURNAL OF ANTIBIOTICS
·
2021
·Vol. 74
·期 5
·330-336
·
IF 2.649
Lan Sugui,Li Zhirong,Su Aiqiu,Peng Yanhong,Liao Yanke,Liu Xuemei,Tan Qiang
The purpose of this study was focused on the mechanisms of the cross-resistance to tetracycline (TET), piperacillin Sodium (PIP), and gentamicin (GEN) in Staphylococcus aureus (SA) mediated by Rhizoma Coptidis extracts (RCE). The selected strains were exposed continuously to RCE at the sublethal concentrations for 12 days, respectively. The susceptibility change of the drug-exposed strains was determined by analysis of the minimum inhibitory concentration. The 16S rDNA sequencing method was used to identify the RCE-exposed strain. Then the expression of resistant genes in the selected isolates was analyzed by transcriptome sequencing. The results indicated that RCE could trigger the preferential cross-resistance to TET, PIP, and GEN in SA. The correlative resistant genes to the three kinds of antibiotics were upregulated in the RCE-exposed strain, and the mRNA levels of the resistant genes determined by RT-qPCR were consistent with those from the transcriptome analysis. It was suggested from these results that the antibacterial Traditional Chinese Medicines might be a significant factor of causing the bacterial antibiotic-resistance.
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JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY
·
2019
·
IF 4.067
Sai Kumar Tammina,Yang Wan,Yanyan Li,Yaling Yang
A simple and one-step microwave digestion method was utilized to synthesize the highly photoluminescent glucosamine derived nitrogen and zinc doped carbon dots (N, Zn-CDs) with a bluish-green luminescence property. The synthesized N, Zn-CDs inherited a good photoluminescence property with a quantum yield of 74% and which were characterized by XPS , XRD , TEM , FT-IR, UV–Vis, and fluorescence spectrometry . More importantly, N, Zn-CDs served as a fluorescence sensor for the detection of Fe 3+ ions, under optimal conditions, a good linear relationship was established in the concentration range of 0.25–125 μM. Also, the experimental results showed the detection limit of N, Zn-CDs against Fe 3+ was 0.15 μM, this could be a promising assay for sensing Fe 3+ ions. Also, the role of zinc doping was explored by conducting the detection under similar experimental conditions in the absence of zinc, and reports revealed that the reason for high photoluminescence was due to the zinc doping. Along with the sensing application, our group had investigated the bactericidal property of synthesized N, Zn-CDs against gram-negative ( Escherichia coli ), and gram-positive pathogens ( Staphylococcus aureus ). N, Zn-CDs displayed good bactericidal activity against both pathogens under light conditions, but Escherichia coli , even under the dark condition proved its activity. Besides, the morphology of both the strains was displayed before and after the treatment via a scanning electron microscopic images. Also, a plausible mechanism for bactericidal activity was also explained. The reported synthesis method could provide a novel approach for the design of high photoluminescent N, Zn–CDs with good sensing and bactericidal property towards Fe 3+ ions, gram-negative and positive bacterial strains .
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ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2025
·
IF 3.8
He Dongxia,Liu Qi,Wang Lei,Han Qinqin,Zhang Jinyang,Li Chao,Song Yuzhu
A novel strategy integrating Brevinin-1BW (BW)-functionalized magnetic bead-based separation with a colorimetric immunoassay was developed to concentrate and enrich Staphylococcus aureus ( S. aureus ) from sample matrices using magnetic beads functionalized with antimicrobial peptide BW (MBs-BW) as carriers. To ensure the selectivity of the method, horseradish peroxidase (HRP)-labeled porcine IgG (HRP-porcine IgG) was used as a molecular recognition reagent and signal amplification probe. MBs-BW/ S. aureus /HRP-porcine IgG probe sandwich complex was successfully obtained. HRP chromogenic substrate 3,3′,5,5′-tetramethylbenzidine (TMB) was used to realize the chromogenic reaction. Finally, S. aureus was quantitatively analyzed based on the color signal generated. Under the optimal conditions, the assay showed a response in the concentration range of 1.0 × 10 2 –1.0 × 10 7 CFU/mL of S. aureus , and the detection limit was as low as 60 CFU/mL. This method does not require complicated sample pretreatment, and the operation process is fast and simple, which can directly complete the whole process of bacterial enrichment and detection. The recovery of this strategy in different types of spiked samples reached 86.8–97.8%, and the analytical results of this strategy were highly consistent with those of the conventional plate counting method. This strategy provides a new way for the rapid detection of S. aureus in complex samples. Graphical
研究领域:
微生物学
·
生物化学
·
分析化学
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