FOOD RESEARCH INTERNATIONAL
·
2023
·
IF 8.1
Danliangmin Song,Ai Jia,Xuehe Qi,Kai Dong,Shiyu Liu,Chaoxin Man,Xinyan Yang,Yujun Jiang
Bacterial biofilm is a protective matrix composed of metabolites secreted by bacteria that envelop bacteria. By forming a biofilm, bacteria can considerably improve their environmental tolerance. In food-related processing environment, different types of microorganisms are often present in biofilms. The main contaminating strain in the powdered infant formula (PIF) processing environment, Cronobacter sakazakii and Staphylococcus aureus continues to pollute the PIF processing environment after biofilm production. This study selected Cronobacter sakazakii with a weak biofilm-forming ability as one of the test organisms. The coexistence of Cronobacter sakazakii and Staphylococcus aureus on the surface of production equipment was simulated to analyze the interaction. Biofilm formation in the co-culture group was significantly higher than the others. In-depth study of the effect of Staphylococcus aureus on the biofilm formation genes of Cronobacter sakazakii . Results show two bacteria can coexist on the surface of a metal device, forming a more compact hybrid biofilm structure. Under co-culture conditions, S. aureus increased bcsA and fliD expression in Cronobacter sakazakii , whereas decreased bcsC expression. Signaling molecules produced by Staphylococcus aureus (Autoinducer 2) significantly promoted the biofilm formation of Cronobacter sakazakii at the concentration of 0–500 ng/mL (0.099–0.177) and up-regulated the expression of bcsA , filD and flhD genes.
研究领域:
微生物学
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食品科学
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PubMed
高影响因子
Scientific Reports
·
2025
·Vol. 15
·期 1
·1-13
·
IF 3.9
Xue Juan,Meng Kun,Lv Jun,Liu Lanfang,Duan Fangfang,Ji Xuemeng,Ding Li
Cronobacter sakazakii is a Gram-negative bacterium known for causing severe infections in neonates, particularly through contaminated infant formula. This study investigated the role of the outer membrane lipoprotein NlpD in the environmental tolerance of C. sakazakii . A nlpD knockout mutant was constructed, and its impact on desiccation resistance, biofilm formation, motility, and proteomic profiles was evaluated and compared with that of the wild-type strain. The nlpD mutant presented reduced desiccation tolerance, reduced ability to form a biofilm, and altered surface hydrophobicity and motility patterns. The complemented strain restored these phenotypic changes, confirming that the observed effects were specifically due to the deletion of nlpD . Proteomic analysis revealed significant differential expression of proteins involved in metabolic and biosynthetic pathways upon nlpD deletion. These findings emphasize the multifaceted role of NlpD in enhancing the environmental tolerance of C. sakazakii , suggesting its importance in the resilience and survival of the bacterium in adverse conditions.
研究领域:
微生物学
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分子生物学
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PubMed
高影响因子
Analytical Methods
·
2017
·Vol. 9
·期 44
·6286-6292
·
IF 1.9
Lingxian Ye,Guangying Zhao,Wenchao Dou
Developing portable and low-cost methods for quantitative detection of Cronobacter sakazakii (C. sakazakii) from powdered infant formula plays a significant role in controlling and preventing a range of serious neonatal infections. Here, we reported a novel methodology for C. sakazakii detection using a low-cost, simple, and widely accessible personal glucose meter (PGM) as the readout. The electrochemical immunoassay strategy was based on antibody and glucose oxidase coated silica nanoparticles (SiNP–GOX–IgG) and antibody functionalized silica-coated magnetic nanoparticles (MNP–IgG). In the process of detecting a positive sample, a sandwich immunocomplex was formed between the MNP–IgG, C. sakazakii and SiNP–GOX–IgG. Then, the above sandwich composites were dispersed in glucose solution, and the concentration of glucose was measured using the PGM. Under optimal conditions, a linear relationship between the decrease of glucose concentration and the logarithm of C. sakazakii concentration was obtained in the concentration range from 9.0 × 102 to 9.0 × 107 CFU mL−1 with a detection limit of 4.2 × 101 CFU mL−1 (S/N = 3). This simple immunoassay holds great potential as an efficient, sensitive and selective point-of-care testing (POCT) tool for C. sakazakii detection.
研究领域:
微生物学
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分析化学
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医学诊断
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DOI
高影响因子
FOOD MICROBIOLOGY
·
2025
·
IF 4.6
Ping Lu,Juan Xue,Aiying Shi,Xuemeng Ji
Cronobacter sakazakii is a notorious foodborne pathogen linked to severe infections in neonates, with powdered infant formula serving as a common transmission route due to its ability to survive under desiccated conditions. Understanding the molecular mechanisms underlying its desiccation resistance is crucial for developing effective control strategies. In this study, we employed comparative proteomics and hydropathy profiling to investigate putative core proteins potentially involved in desiccation resilience. Proteomic analysis of wild-type and 22 knockout strains identified 1,354 proteins, of which 156 were defined as putative core proteins due to stable abundance profiles. Among these, two uncharacterized proteins, ESA_03998 and ESA_01764 , exhibited strong hydrophilic properties and were predicted to be surface-associated based on signal peptide analysis. Gene knockout of ESA_03998 or ESA_01764 significantly reduced surface hydrophilicity and survival under prolonged desiccation in both PBS and milk powder matrices. Additionally, Laurdan-based membrane fluidity assays revealed increased membrane rigidification (ΔGP) in mutant strains, indicating that these proteins help preserve membrane hydration and dynamics under low-moisture conditions. These findings establish a novel link between hydrophilic putative core proteins and membrane-associated desiccation resistance in C. sakazakii , offering insights into its persistence in dry food environments and highlighting potential targets for mitigating contamination in powdered infant formula.
研究领域:
微生物学
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分子生物学
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食品科学
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PubMed
高影响因子
FOOD RESEARCH INTERNATIONAL
·
2018
·
IF 3.52
Yong Chen,Yifan Zhang,Xiaohou Wang,Jianqun Ling,Guanghua He,Lirong Shen
Cronobacter sakazakii ( Cs ) is a typical foodborne bacterium that infect powdered infant formula (PIF) worldwide. In this study, a recombinant antimicrobial peptide, branded as Funme peptide (FP)was applied to protect PIF from Cs contamination. The result from the antimicrobial activity assay showed that the minimum inhibitory concentration (MIC) of BMAP-27 peptide, FP and Ampicillin against Cs were 250.0, 125.0 and 15.6 μg/mL, respectively, indicating FP possessed higher MIC than that of Ampicillin, and lower MIC than that of BMAP-27. The minimum biofilm eradication concentration (MBEC) assay showed that FP at 2 × MIC (250.0 μg/mL) could completely eradicated Cs biofilms. The antibacterial activity of FP might be due to the increasing permeability and the release of cytoplasmic β-galactosidase of Cs . The results acquired from transmission electron microscopy and scanning electron microscopy indicated that FP induced the disruption and dysfunction of cell walls and membranes. Moreover, safety assay showed that FP had low cytotoxicity to human erythrocytes. The present study investigated the antibacterial effects and mechanisms of FP against Cs , providing promising evidence to apply this novel antimicrobial agent against Cs contamination in foods and food processing facilities.
研究领域:
微生物学
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食品安全
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分子生物学
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PubMed
高影响因子
Frontiers in Microbiology
·
2022
·
IF 6.064
Lu Ping,Ji Xuemeng,Xue Juan,Dong Yinping,Chen Xi
Screening drugs and compounds to fight against Cronobacter sakazakii (C. sakazakii), one of the most common pathogens that can cause fatal necrotizing enterocolitis, septicema and meningitis, is still needed. We found that p-coumaric acid (pCA) has an inhibitory effect on C. sakazakii in vitro and in vivo. Proteomic changes of C. sakazakii BAA-894 exposed to pCA were studied to reveal the antibacterial mechanisms involved. A total of 1,553 proteins were identified in C. sakazakii BAA-894 by label-free proteomics analysis. Fuzzy cluster analysis showed that 33 were up-regulated, and 110 were down-regulated with pCA treatment. Gene Ontology (GO) analysis concluded that pCA caused the change of metabolic state of bacteria and generally in the state of metabolic inhibition. KEGG Enrichment Analysis (KEGG) analysis showed that pCA inhibited energy metabolism and distorted the balance of amino acid metabolism. Posttranslational modification analysis showed that pCA affected the deamidation of three proteins, including Elongation factor Tu, one of the vital proteins in bacteria. Molecular docking suggested the hydrogen bond between the pCA carboxyl group and Elongation factor Tu Asn-64 might contribute to deamidation. Overall, we found that pCA interfered with cellular energy and amino acid metabolism and promoted elongation factor Tu deamidation, suggesting that pCA can be an effective natural substitute to control C. sakazakii.
研究领域:
微生物学
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蛋白质组学
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医学
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PubMed
高影响因子
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY
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2023
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IF 5.4
Yang Wang,Na Ling,Yaping Wang,Dexin Ou,Zhao Liang,Guoqing Li,Hongyuan Zhao,Yingwang Ye
Cronobacter sakazakii ( C. sakazakii ) is a common food-borne pathogen that induces meningitis, sepsis, and necrotizing enterocolitis, primarily in newborns and infants. Iron plays a pivotal role in the growth of cells and biofilm formation. However, the effects of hemin (ferric ion donor) on C. sakazakii cells are scarcely known. Here, we explored the effect of ferric ions on the growth of planktonic C. sakazakii , biofilm formation, and swarming motility by crystal violet staining (CVS), scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), and swarming assay. Our study demonstrated that ferric ions facilitated the growth of planktonic C. sakazakii , while hemin at concentrations ranging from 50 to 800 μmol/L promoted biofilm formation and at concentrations between 50 and 200 μmol/L enhanced the swarming motility of C. sakazakii . Furthermore, high hemin concentrations (400–800 μmol/L) were found to reduce flagellar length, as confirmed by transmission electron microscopy (TEM). These findings indicated that ferric ions mediated the swarming motility of C. sakazakii by regulating flagellar assembly. Finally, transcriptomic analysis of C. sakazakii was performed at hemin concentrations of 0, 50, and 200 μmol/L, which revealed that several genes associated with iron transport and metabolism, and flagellar assembly were essential for the survival of C. sakazakii under hemin treatment. Our findings revealed the molecular basis of ferric ions on C. sakazakii growth and biofilm formation, thus providing a novel perspective for its prevention and control.
研究领域:
微生物学
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食品安全
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分子生物学
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PubMed
高影响因子
JOURNAL OF FOOD SCIENCE
·
2026
·Vol. 91
·期 7
·e71285
·
IF 4.1
Yu Zheng,Shiqin Wu,Yujia Sun,Li Zhou,Xiangmei Ren,Yuanhong Li,Fuxing Lin
Cronobacter sakazakii is a significant foodborne pathogen commonly associated with dairy products, causing severe infections with high mortality rates, particularly in neonates and immunocompromised adults. In this study, we demonstrated that apigenin exhibits potent antibacterial activity against C. sakazakii , with a minimum inhibitory concentration (MIC) of 160 mg/L. Propidium iodide staining indicated that apigenin at sub-MIC and MIC concentrations had negligible effects on bacterial membrane integrity; only a 4.9% increase in membrane permeability was observed at 2 MIC, suggesting that membrane damage is not the primary antibacterial mechanism. Further analysis revealed that apigenin treatment significantly reduced Adenosine triphosphate (ATP) levels within 2 h without altering growth kinetics or membrane integrity, indicating that ATP reduction resulted from metabolic disruption rather than cell death. Targeted metabolomics analysis revealed distinct separation in metabolic profiles between apigenin-treated and control groups, identifying 19 differential metabolites enriched in multiple pathways, with the citric acid (TCA) cycle being the most profoundly perturbed pathway. Biochemical assays confirmed that apigenin markedly decreased the activities of four key TCA cycle enzymes: isocitrate dehydrogenase (ICDHc, 40.09% reduction), α-ketoglutarate dehydrogenase (α-KGDH, 39.26% reduction), succinate dehydrogenase (SDH, 97.66% reduction), and malate dehydrogenase (MDH, 85.86% reduction). Molecular docking further supported the potential interaction between apigenin and these TCA cycle-related enzymes. Collectively, our findings suggest that apigenin inhibits C. sakazakii growth in association with the inhibition of key TCA cycle enzymes, which may contribute to the disruption of central carbon metabolism and ATP biosynthesis, ultimately leading to bacterial metabolic dysfunction and growth inhibition.
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高影响因子
Microbiology Spectrum
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2025
·
IF 3.8
Yue-yue Zhang,Ding-rong Zhang,Zhen-quan Yang,Lei Yuan,Ya Zhao,Yuan-song Zhang
Cronobacter sakazakii is an opportunistic pathogen that can contaminate infant formula, resulting in meningitis and neurological disorders in infants. Bacteriophages have been consistently identified as effective inhibitors of pathogenic bacteria. In this study, a lytic phage, Csp-D17, was isolated from environmental wastewater, and its characterization, genomics, and application were analyzed. Transmission electron microscopy revealed that the phage belonged to the order Caudoviricetes within the family Ackermannviridae. The phage Csp-D17 exhibited notable stability across varied thermal and pH conditions, with an optimal multiplicity of infection of 0.01 and a lysis capacity of 40 plaque-forming units per infected cell. Evaluation of its host range revealed lytic activity against 4 out of 11 tested strains of C. sakazakii (36.4%). The frequency of insensitive mutations in C. sakazakii D17 to phage Csp-D17 was 3.8 × 10−7. High-dose phage treatment was more effective in inhibiting the growth of C. sakazakii D17 in the powdered infant formula model, which was correlated with its short latency period, high lysis amount, and low frequency of insensitive mutations in the host bacteria. The results demonstrated that phage Csp-D17 was not lysogenic. Furthermore, the genomic study of phage Csp-D17 revealed that the genome structure of this phage was a double-stranded DNA molecule with a length of 143,081 base pairs, a guanine + cytosine content of 46.42%, 233 open reading frames (the average gene length is 553 base pairs), and it does not contain any genes related to virulence or lysogenicity. Furthermore, the genetic function, expression mechanism, and phylogenetic diversity of Csp-D17 were analyzed by genomic comparison. This study provides a theoretical basis for developing novel biocontrol agents based on phage control of C. sakazakii.
研究领域:
微生物学
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病毒学
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食品安全
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基因组学
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分子生物学
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噬菌体治疗
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DOI
PubMed
高影响因子
VIRUS RESEARCH
·
2023
·
IF 5
Yuan-Song Zhang,Lei Yuan,Fedrick C. Mgomi,Cao-Wei Chen,Yang Wang,Zhen-Quan Yang,Xin-an Jiao
Cronobacter sakazakii, a foodborne pathogen, can contaminate powdered infant formula (PIF) and cause life-threatening meningitis, necrotizing colitis and meningoencephalitis in infants. Bacteriophages are increasingly considered an efficient approach to target pathogenic microorganisms. In the current study, four virulent phages that can infect C. sakazakii were isolated from sewage samples, and their biological and complete genomic characteristics were analyzed. A comparative genomic analysis was performed to investigate the functional genes and phylogenetic evolution of the four phages. The results revealed that all four phages belonged to the Ackermannviridae family. Notably, the viral burst size of the phages ranged from 10 to 250 PFU/cell, following a latent period of 5 min to 20 min. Moreover, phages were stable over a pH range of 4 to 10 and a temperature range of 50 ℃ to 60 ℃. The full length of the complete genomes of the four phages ranged from 41,929 bp to 146,806 bp, containing lysis genes but no virulence genes. Phylogenetic tree analysis showed that the four phages were members of two distinct genetic groups with a significant genetic evolutionary distance between each C. sakazakii phage. Furthermore, the antibacterial assay revealed that all phages could inhibit the growth of C. sakazakii for up to 24 h. Taken together, the four phages have huge prospects as additives in dairy products to counter C. sakazakii .
研究领域:
微生物学
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基因组学
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食品安全
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分子生物学
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DOI
PubMed
高影响因子
FOOD RESEARCH INTERNATIONAL
·
2025
·
IF 8
Aiying Shi,Juan Xue,Yongshuai Liu,Jianhui Wang,Xiaofei Liu,Lianqiang Song,Shanming Sun
Cronobacter sakazakii is a Gram-negative, foodborne pathogen associated with severe infections, particularly in infants and immunocompromised individuals. Given its significant public health implications and its remarkable resilience in powdered infant formula, understanding the genetic factors that contribute to its virulence and environmental tolerance is crucial. This study focuses on the kdpE gene in Cronobacter sakazakii , part of a two-component regulatory system known for its role in adaptation to environmental stress. A kdpE knockout mutant of Cronobacter sakazakii was constructed to evaluate its effects on bacterial growth, virulence, biofilm formation, and desiccation tolerance. The results revealed that kdpE deletion reduced bacterial growth and virulence, as evidenced by higher survival rates in rats infected with the Δ kdpE mutant compared to the wild-type strain. Notably, the Δ kdpE mutant showed significant downregulation of proteins involved in carbon metabolism and respiratory activity, while proteomic analysis revealed increased deamidation of proteins related to glycolysis. Furthermore, biofilm formation was significantly impaired in the mutant, although its desiccation tolerance was enhanced, attributed to the upregulation of the molecular chaperone GrpE. These findings suggest that kdpE plays a crucial role in regulating the virulence and environmental adaptability of Cronobacter sakazakii , with implications for understanding its pathogenic mechanisms and developing preventive measures.
研究领域:
微生物学
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分子生物学
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遗传学
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发病机制
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食品微生物学
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DOI
PubMed
高影响因子
FOOD RESEARCH INTERNATIONAL
·
2023
·
IF 8.1
Juan Xue,Jun Lv,Lanfang Liu,Fangfang Duan,Aiying Shi,Xuemeng Ji,Li Ding
Cronobacter sakazakii ( C. sakazakii ) is a notorious pathogen responsible for infections in infants and newborns, often transmitted through contaminated infant formula. Despite the use of traditional pasteurization methods, which can reduce microbial contamination, there remains a significant risk of pathogenic C. sakazakii surviving due to its exceptional stress tolerance. In our study, we employed a comparative proteomic approach by comparing wild-type strains with gene knockout strains to identify the essential genes crucial for the successful survival of C. sakazakii during desiccation. Our investigation revealed the significance of envZ-ompR , recA , and flhD gene cassettes in contributing to desiccation tolerance in C. sakazakii . Furthermore, through our comparative proteomic profiling, we identified the maltodextrin-binding protein encoded by ESA_03421 as a potential factor influencing dry tolerance. This protein is regulated by EnvZ-OmpR , RecA , and FlhD . Notably, the knockout of ESA_03421 resulted in a 150% greater reduction in Log CFU compared to the wild-type C. sakazakii . Overall, our findings offer valuable insights into the mechanisms underlying C. sakazakii desiccation tolerance and provide potential targets for the development of new antimicrobial strategies aimed at reducing the risk of infections in infants and newborns.
研究领域:
微生物学
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蛋白质组学
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食品安全
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婴儿健康
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DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）