FOOD RESEARCH INTERNATIONAL
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2023
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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.
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高影响因子
JOURNAL OF FOOD SCIENCE
·
2026
·Vol. 91
·期 7
·e71285
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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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Microbiology Spectrum
·
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.
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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 .
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INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY
·
2023
·
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.
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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.
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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.
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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.
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APPLIED AND ENVIRONMENTAL MICROBIOLOGY
·
2023
·Vol. 89
·期 7
·
IF 4.4
Ping Lu,Juan Xue,Xi Chen,Xuemeng Ji
Cronobacter sakazakii is a Gram-negative bacterium that causes infections in individuals of all ages, with neonates being the most vulnerable group. The objective of this study was to explore the function of the dnaK gene in C. sakazakii and to elucidate the impact of alterations in the protein composition regulated by dnaK on virulence and stress adaptation. Our research demonstrates the critical role of the dnaK gene in various key virulence factors, including adhesion, invasion, and acid resistance in C. sakazakii. Through the use of proteomic analysis, we discovered that deletion of the dnaK gene in C. sakazakii leads to an upregulation of protein abundance and increased levels of deamidated posttranscriptional modifications, suggesting that DnaK may play a role in maintaining proper protein activity by reducing protein deamidation in bacteria. These findings indicate that DnaK-mediated protein deamidation may be a novel mechanism for virulence and stress adaptation in C. sakazakii. These findings suggest that targeting DnaK could be a promising strategy for developing drugs to treat C. sakazakii infections.IMPORTANCE Cronobacter sakazakii can cause disease in individuals of all ages, with infections in premature infants being particularly deadly and resulting in bacterial meningitis and sepsis with a high mortality rate. Our study demonstrates that dnaK in Cronobacter sakazakii plays a critical role in virulence, adhesion, invasion, and acid resistance. Using proteomic analysis to compare protein changes in response to dnaK knockout, we found that dnaK knockout significantly upregulates the abundance of some proteins but also results in the deamidation of many proteins. Our research has identified a connection between molecular chaperones and protein deamidation, which suggests a potential future drug development strategy of targeting DnaK as a drug target.
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2024
·
IF 5.7
Xuemeng Ji,Aiying Shi,Jin Wang,Bowei Zhang,Yaozhong Hu,Huan Lv,Jing Wu,Yi Sun,Jing-Min Liu,Yan Zhang,Shuo Wang
Cronobacter sakazakii is a notorious foodborne opportunistic pathogen, particularly affecting vulnerable populations such as premature infants, and poses significant public health challenges. This study aimed to elucidate the role of the envZ/ompR genes in environmental tolerance, pathogenicity, and protein regulation of C. sakazakii. An envZ/ompR knockout mutant was constructed and assessed for its impact on bacterial growth, virulence, environmental tolerance, and protein regulation. Results demonstrate that deletion of envZ/ompR genes leads to reduced growth rate and attenuated virulence in animal models. Additionally, the knockout strain exhibited compromised environmental tolerance, particularly in desiccation and oxidative stress conditions, along with impaired adhesion and invasion abilities in epithelial cells. Proteomic analysis revealed significant alterations in protein expression and phosphorylation patterns, highlighting potential compensatory mechanisms triggered by gene deletion. Furthermore, investigation into protein deamidation and glucose metabolism uncovered a link between envZ/ompR deletion and energy metabolism dysregulation. Interestingly, the downregulation of MalK and GrxC proteins was identified as contributing factors to altered desiccation tolerance and disrupted redox homeostasis, respectively, providing mechanistic insights into the phenotypic changes observed. Overall, this study enhances understanding of the multifaceted roles of envZ/ompR in C. sakazakii physiology and pathogenesis, shedding light on potential targets for therapeutic intervention and food safety strategies.
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JOURNAL OF DAIRY SCIENCE
·
2021
·
IF 4.034
Siyuan Gao,Jinhui Wu,Hong Wang,Shengying Hu,Li Meng
ABSTRACT The presence of Cronobacter sakazakii must be controlled in infant powder plants, because it may cause infectious disease in infants, with high mortality. Testing for C. sakazakii in powdered infant formula should be performed before delivery, and it requires rapid and specific detection methods. In this study, we established a surface-enhanced Raman scattering (SERS) immunochromatographic test strip for the quantitative determination of C. sakazakii in powdered infant formula. Monoclonal antibodies for C. sakazakii were labeled with p -aminothiophenol-bound colloidal gold nanoparticles. Color change in the test line indicated the presence of C. sakazakii . A highly sensitive and quantitative test method was developed based on the Raman signal produced by the p -aminothiophenol bonding on gold nanoparticles. The SERS immunochromatographic test strip assay required a short analysis time (12 min) and exhibited a linearity range from 10 2 to 10 7 cfu/mL. The limit of detection was 201 cfu/mL without preculture. The SERS immunochromatographic test strip assay is a promising tool for the simple and rapid quantitative analysis of C. sakazakii and other pathogenic bacteria.
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高影响因子
RESEARCH IN MICROBIOLOGY
·
2025
·
IF 3.4
Ping Lu,Juan Xue,Aiying Shi,Xuemeng Ji
Purpose Cronobacter sakazakii is a foodborne pathogen notable for both virulence and desiccation tolerance. This study investigates the functional roles of the pESA3 plasmid in mediating these two traits and explores the potential trade-off between them. Methods Proteomic profiling identified pESA3 -encoded proteins, followed by construction of targeted deletion mutants ( hcp , ESA_pESA3p05542 ) and a plasmid-cured strain. Phenotypic assays evaluated growth, desiccation survival, surface hydrophilicity, and virulence in a neonatal rat model. Results Deletion of ESA_pESA3p05542 impaired desiccation tolerance and reduced surface hydrophilicity, while hcp deletion attenuated virulence but enhanced desiccation survival. Notably, complete loss of pESA3 abolished virulence without affecting desiccation resistance, suggesting functional antagonism between plasmid-encoded virulence and stress-response genes. Conclusions pESA3 enables C. sakazakii to balance host pathogenicity and environmental persistence. Its gene modules promote either virulence or desiccation resistance, but not both, reflecting an evolutionary trade-off that supports survival across diverse niches.
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