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1.
Int J Food Microbiol ; 426: 110925, 2025 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-39366090

RESUMO

Biofilm formation of Staphylococcus aureus in food processing environments raises significant safety concerns, necessitating the development of new antibiofilm approaches for controlling S. aureus contamination. This study aimed to elucidate the antibiofilm mechanism of 2R,3R-dihydromyricetin (DMY), a natural flavonoid, against S. aureus and evaluate its efficacy in reducing bacterial adhesion to eggshell. The results revealed that DMY was a potent inhibitor of S. aureus sortase A (SrtA) with an IC50 of 73.43 µM, preventing bacterial adhesion to fibrinogen and subsequent biofilm formation. Fluorescence quenching assay and surface plasmon resonance analysis confirmed that DMY could directly bind to S. aureus SrtA. Notably, circular dichroism spectra demonstrated a conformational change in SrtA from α-helical to ß-sheet structure upon DMY binding. Molecular dynamics simulation suggested that DMY bound to the catalytic pocket of S. aureus SrtA via hydrophobic interactions and hydrogen bonds. Furthermore, fluorescence microscopic observations further revealed that DMY attenuated the biofilm-related phenotype of SrtA by decreasing the anchoring of S. aureus protein A (SpA) onto cell wall. Importantly, pretreatment with 125 µg/mL DMY significantly reduced 1.14-1.75 log CFU/cm2 of S. aureus adhered on eggshells. Overall, these findings highlight how specific targeting of SrtA by DMY inhibits the attachment stages of biofilm development in S. aureus, making it a promising candidate for a novel disinfectant against this pathogen in the food industry.


Assuntos
Aminoaciltransferases , Aderência Bacteriana , Proteínas de Bactérias , Biofilmes , Cisteína Endopeptidases , Casca de Ovo , Flavonóis , Staphylococcus aureus , Staphylococcus aureus/efeitos dos fármacos , Cisteína Endopeptidases/metabolismo , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Aminoaciltransferases/metabolismo , Proteínas de Bactérias/metabolismo , Flavonóis/farmacologia , Animais , Aderência Bacteriana/efeitos dos fármacos , Casca de Ovo/microbiologia , Antibacterianos/farmacologia , Simulação de Dinâmica Molecular , Galinhas
2.
Int J Food Microbiol ; 426: 110874, 2025 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-39244811

RESUMO

Biofilms are a critical factor for food safety, causing important economic losses. Among the novel strategies for controlling biofilms, essential oils (EOs) can represent an environmentally friendly approach, able to act both on early and mature stages of biofilm formation. This review reports the anti-biofilm mechanisms of action of EOs against five pathogenic bacterial species known for their biofilm-forming ability. These mechanisms include disturbing the expression of genes related to quorum sensing (QS), motility, adhesion, and virulence. Biofilms and QS are interconnected processes, and EOs interfere with the communication system (e.g. regulating the expression of agrBDCA, luxR, luxS, and pqsA genes), thus influencing biofilm formation. In addition, QS is an important mechanism that regulates gene expression related to bacterial survival, virulence, and pathogenicity. Similarly, EOs also influence the expression of many virulence genes. Moreover, EOs exert their effects modulating the genes associated with bacterial adhesion and motility, for example those involved in curli (csg), fimbriae (fim, lpf), and flagella (fla, fli, flh, and mot) production, as well as the ica genes responsible for synthetizing polysaccharide intercellular adhesin. This review provides a comprehensive framework on the topic for a better understanding of EOs biofilm mechanisms of action.


Assuntos
Aderência Bacteriana , Biofilmes , Óleos Voláteis , Percepção de Quorum , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Percepção de Quorum/efeitos dos fármacos , Óleos Voláteis/farmacologia , Virulência , Aderência Bacteriana/efeitos dos fármacos , Bactérias/efeitos dos fármacos , Bactérias/patogenicidade , Bactérias/genética , Antibacterianos/farmacologia , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos
3.
Int J Food Microbiol ; 426: 110913, 2025 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-39293097

RESUMO

This study investigated the influence of food contact surface materials on the biofilm formation of Vibrio parahaemolyticus while attempting to minimize the impact of environmental factors. The response surface methodology (RSM), incorporating three controlled environmental factors (temperature, pH, and salinity), was employed to determine the optimal conditions for biofilm formation on stainless steel (SS) and polypropylene (PP) coupons. The RSM results demonstrated that pH was highly influential. After minimizing the impacts of environmental factors, initially V. parahaemolyticus adhered more rapidly on PP than SS. To adhere to SS, V. parahaemolyticus formed extra exopolysaccharide (EPS) and exhibited clustered stacking. Both PP and SS exhibited hydrophilic properties, but SS was more hydrophilic than PP. Finally, this study observed a higher transfer rate of biofilms from PP to fish fillets than from SS to fish fillets. The present findings suggest that the food industry should consider the material of food processing surfaces to prevent V. parahaemolyticus biofilm formation and thus to enhance food safety.


Assuntos
Biofilmes , Polipropilenos , Aço Inoxidável , Vibrio parahaemolyticus , Vibrio parahaemolyticus/crescimento & desenvolvimento , Vibrio parahaemolyticus/fisiologia , Biofilmes/crescimento & desenvolvimento , Animais , Microbiologia de Alimentos , Concentração de Íons de Hidrogênio , Aderência Bacteriana , Alimentos Marinhos/microbiologia , Peixes/microbiologia , Temperatura , Contaminação de Alimentos/prevenção & controle , Contaminação de Alimentos/análise
4.
FASEB J ; 38(20): e70123, 2024 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-39436142

RESUMO

Tissue injury, including extracellular matrix (ECM) degradation, is a hallmark of group A Streptococcus (GAS) skin infection and is partially mediated by M proteins which possess lectin-like properties. Hyaluronic acid is a glycosaminoglycan enriched in the cutaneous ECM, yet an interaction with M proteins has yet to be explored. This study revealed that hyaluronic acid binding was conserved across phylogenetically diverse M proteins, mediated by RR/SR motifs predominantly localized in the C repeat region. Keratinocyte wound healing was decreased through the recruitment of hyaluronic acid by M proteins in an M type-specific manner. GAS strains 5448 (M1 serotype) and ALAB49 (M53 serotype) also bound hyaluronic acid via M proteins, but hyaluronic acid could increase bacterial adherence independently of M proteins. The identification of host-pathogen mechanisms that affect ECM composition and cell repair responses may facilitate the development of nonantibiotic therapeutics that arrest GAS disease progression in the skin.


Assuntos
Ácido Hialurônico , Streptococcus pyogenes , Ácido Hialurônico/metabolismo , Streptococcus pyogenes/metabolismo , Humanos , Antígenos de Bactérias/metabolismo , Proteínas da Membrana Bacteriana Externa/metabolismo , Queratinócitos/metabolismo , Queratinócitos/microbiologia , Proteínas de Transporte/metabolismo , Infecções Estreptocócicas/metabolismo , Infecções Estreptocócicas/microbiologia , Motivos de Aminoácidos , Cicatrização , Serina/metabolismo , Aderência Bacteriana , Arginina/metabolismo , Ligação Proteica
5.
Cell Mol Life Sci ; 81(1): 435, 2024 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-39412594

RESUMO

Staphylococcus aureus is a pathogen associated with severe respiratory infections. The ability of S. aureus to internalize into lung epithelial cells complicates the treatment of respiratory infections caused by this bacterium. In the intracellular environment, S. aureus can avoid elimination by the immune system and the action of circulating antibiotics. Consequently, interfering with S. aureus internalization may represent a promising adjunctive therapeutic strategy to enhance the efficacy of conventional treatments. Here, we investigated the host-pathogen molecular interactions involved in S. aureus internalization into human lung epithelial cells. Lipid raft-mediated endocytosis was identified as the main entry mechanism. Thus, bacterial internalization was significantly reduced after the disruption of lipid rafts with methyl-ß-cyclodextrin. Confocal microscopy confirmed the colocalization of S. aureus with lipid raft markers such as ganglioside GM1 and caveolin-1. Adhesion of S. aureus to α5ß1 integrin on lung epithelial cells via fibronectin-binding proteins (FnBPs) was a prerequisite for bacterial internalization. A mutant S. aureus strain deficient in the expression of alpha-hemolysin (Hla) was significantly impaired in its capacity to enter lung epithelial cells despite retaining its capacity to adhere. This suggests a direct involvement of Hla in the bacterial internalization process. Among the receptors for Hla located in lipid rafts, caveolin-1 was essential for S. aureus internalization, whereas ADAM10 was dispensable for this process. In conclusion, this study supports a significant role of lipid rafts in S. aureus internalization into human lung epithelial cells and highlights the interaction between bacterial Hla and host caveolin-1 as crucial for the internalization process.


Assuntos
Caveolina 1 , Colesterol , Endocitose , Células Epiteliais , Proteínas Hemolisinas , Pulmão , Microdomínios da Membrana , Staphylococcus aureus , Humanos , Staphylococcus aureus/metabolismo , Microdomínios da Membrana/metabolismo , Proteínas Hemolisinas/metabolismo , Caveolina 1/metabolismo , Colesterol/metabolismo , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Pulmão/metabolismo , Pulmão/microbiologia , Toxinas Bacterianas/metabolismo , Interações Hospedeiro-Patógeno , beta-Ciclodextrinas/farmacologia , Aderência Bacteriana , Integrina alfa5beta1/metabolismo , Infecções Estafilocócicas/metabolismo , Infecções Estafilocócicas/microbiologia , Células A549 , Proteína ADAM10/metabolismo
6.
Sci Rep ; 14(1): 24549, 2024 10 19.
Artigo em Inglês | MEDLINE | ID: mdl-39426980

RESUMO

This study is the first one that investigates in detail the bacterial intercellular response to the high density of crystallographic defects including vacancies created in Cu by high pressure torsion. To this aim, samples were deformed by high pressure torsion and afterward, their antibacterial properties against Staphylococcus aureus were analyzed in adhesion tests. As a reference an annealed sample was applied. To avoid the influence of surface roughness, specially elaborated conditions for surface preparation were employed, which do not introduce defects and assure comparable surface roughness. The analysis of the chemical composition and thickness of passive layers by X-ray photoelectron spectroscopy showed that they were comparable for nanostructured and micrograined samples, consisting of Cu2O and CuO, and a thickness of 6 nm. The interface bacterium-substrate was prepared by a focused ion beam and further analyzed by scanning transmission electron microscopy and energy dispersive spectroscopy. High pressure torsion processed Cu shows enhanced anti-adhesion properties while in contact with S. aureus than micrograined Cu. There is a linear correlation between luminous intensity and grain size-0.5. The bacterial intercellular defence mechanism includes the creation of Cu2O nanoparticles and the increased concentration of sulphur-rich compounds near these nanoparticles.


Assuntos
Aderência Bacteriana , Cobre , Staphylococcus aureus , Staphylococcus aureus/efeitos dos fármacos , Aderência Bacteriana/efeitos dos fármacos , Cobre/química , Cobre/farmacologia , Propriedades de Superfície , Espectroscopia Fotoeletrônica , Antibacterianos/farmacologia , Antibacterianos/química
7.
J Nanobiotechnology ; 22(1): 629, 2024 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-39407284

RESUMO

Salmonella Enteritidis is a major foodborne pathogen throughout the world and the increase in antibiotic resistance of Salmonella poses a significant threat to public safety. Natural nanobodies exhibit high affinity, thermal stability, ease of production, and notably higher diversity, making them widely applicable for the treatment of viral and bacterial infections. Recombinant expression using Lactococcus lactis leverages both acid resistance and mucosal colonization properties of these bacteria, allowing the effective expression of exogenous proteins for therapeutic effects. In this study, nine specific nanobodies against the flagellar protein FliC were identified and expressed. In vitro experiments demonstrated that FliC-Nb-76 effectively inhibited the motility of S. Enteritidis and inhibited its adhesion to and invasion of HIEC-6, RAW264.7, and chicken intestinal epithelial cells. Additionally, a recombinant L. lactis strain secreting the nanobody, L. lactis-Nb76, was obtained. Animal experiments confirmed that it could significantly reduce the mortality rates of chickens infected with S. Enteritidis, together with alleviating the inflammatory response caused by the pathogen. These results provide a novel strategy for the treatment of antibiotic-resistant S. Enteritidis infection in the intestinal tract.


Assuntos
Galinhas , Lactococcus lactis , Salmonella enteritidis , Anticorpos de Domínio Único , Lactococcus lactis/genética , Lactococcus lactis/metabolismo , Animais , Camundongos , Anticorpos de Domínio Único/farmacologia , Células RAW 264.7 , Intestinos/microbiologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Humanos , Flagelina/farmacologia , Flagelina/genética , Infecções por Salmonella/microbiologia , Aderência Bacteriana , Linhagem Celular , Salmonelose Animal/microbiologia , Antibacterianos/farmacologia
8.
Vet Res ; 55(1): 138, 2024 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-39407352

RESUMO

The Riemerella anatipestifer bacterium is known to cause infectious serositis in ducklings. Moreover, its adherence to the host's respiratory mucosa is a critical step in pathogenesis. Membrane cofactor protein (MCP; CD46) is a complement regulatory factor on the surface of eukaryotic cell membranes. Bacteria have been found to bind to this protein on host cells. Outer membrane proteins (OMPs) are necessary for adhesion, colonisation, and pathogenicity of Gram-negative bacteria; however, the mechanism by which R. anatipestifer adheres to duck cells remains unclear. In this study, pull-down assays and LC-MS/MS identified eleven OMPs interacting with duck CD46 (dCD46), with OMP71 exhibiting the strongest binding. The ability of an omp71 gene deletion strain to bind dCD46 is weaker than that of the wild-type strain, suggesting that this interaction is important. Further evidence of this interaction was obtained by synthesising OMP71 using an Escherichia coli recombinant protein expression system. Adhesion and invasion assays and protein and antibody blocking assays confirmed that OMP71 promoted the R. anatipestifer YM strain (RA-YM) adhesion to duck embryo fibroblasts (DEFs) by binding to CD46. Tests of the pathogenicity of a Δomp71 mutant strain of RA-YM on ducks compared to the wild-type parent supported the hypothesis that OMP71 was a key virulence factor of RA-YM. In summary, the finding that R. anatipestifer exploits CD46 to bind to host cells via OMP71 increases our understanding of the molecular mechanism of R. anatipestifer invasion. The finding suggests potential targets for preventing and treating diseases related to R. anatipestifer infection.


Assuntos
Aderência Bacteriana , Proteínas da Membrana Bacteriana Externa , Patos , Infecções por Flavobacteriaceae , Proteína Cofatora de Membrana , Doenças das Aves Domésticas , Riemerella , Animais , Riemerella/patogenicidade , Riemerella/genética , Riemerella/metabolismo , Doenças das Aves Domésticas/microbiologia , Proteínas da Membrana Bacteriana Externa/metabolismo , Proteínas da Membrana Bacteriana Externa/genética , Infecções por Flavobacteriaceae/veterinária , Infecções por Flavobacteriaceae/microbiologia , Virulência , Proteína Cofatora de Membrana/metabolismo , Proteína Cofatora de Membrana/genética
9.
Int J Implant Dent ; 10(1): 45, 2024 Oct 17.
Artigo em Inglês | MEDLINE | ID: mdl-39419937

RESUMO

OBJECTIVE: This research endeavors to scrutinize the influence of attachment systems and denture cleaning methodologies on microbial biomass and composition within the realm of implant-supported overdentures, a crucial consideration for patients with dentition defects necessitating such prosthetic solutions. SUBJECTS AND METHODS: Employing five polymethyl methacrylate specimens designed to emulate the fitting surfaces of traditional dentures and implant-supported overdentures. Following the polishing of each specimen and the quantification of its roughness, co-cultivation with three distinct microbial strains ensued, culminating in ultrasonic cleaning in water. The bar-clip group, differentiated by the depth of attachment, underwent cleaning employing four diverse methods. Biomass quantities were meticulously recorded both pre and post cleaning interventions, with subsequent data analysis via t-testing and one-way ANOVA, maintaining a significance level of α = 0.05. RESULTS: The bar-clip groups demonstrated an elevated degree of microbial adhesion, with the deeper locator group exhibiting heightened biomass residue post-cleaning, indicative of increased cleaning complexity. Ultrasonic cleaning predominantly targeted biofilm and deceased bacteria, whereas chemical cleaners primarily reduced the quantity of viable bacteria. The synergistic application of ultrasonics and chemical cleaning treatments yielded the minimal biomass residue. CONCLUSION: In contemplating the utilization of dentures milled by dental computer-aided design/manufacturing systems, meticulous pre-use surface polishing is imperative. The extent of biofilm adhesion correlates with the chosen attachment system. This study advocates for the incorporation of ultrasonic cleaning in conjunction with chemical cleaning solutions to optimize the removal of biofilm and live cellular entities in the context of implant-supported overdentures.


Assuntos
Aderência Bacteriana , Biofilmes , Biomassa , Prótese Dentária Fixada por Implante , Revestimento de Dentadura , Biofilmes/crescimento & desenvolvimento , Aderência Bacteriana/fisiologia , Polimetil Metacrilato/química , Humanos , Propriedades de Superfície , Higienizadores de Dentadura
10.
Int J Mol Sci ; 25(19)2024 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-39408597

RESUMO

Ventilator-associated pneumonia (VAP) remains one of the most common hospital-acquired infections (HAI). Considering the complicated diagnosis and the lack of effective treatment, prophylactic measures are suggested as the new standard to prevent the disease. Although VAP often manifests a polymicrobial nature, Pseudomonas aeruginosa remains one of the pathogens associated with the highest morbidity and mortality rates within these mechanically ventilated patients. In this paper, we report on the development of an antibacterial hydrogel coating using the polymyxin B (PMB) peptide to prevent bacterial adhesion to the polymeric substrate. We fully characterized the properties of the coating using atomic force microscopy (AFM), scanning electron microscopy (SEM), wettability analyses and Fourier-transform infrared (FTIR) and Raman spectroscopy. Furthermore, several biological assays confirmed the antibacterial and anti-biofilm effect of the tubing for at least 8 days against P. aeruginosa. On top of that, the produced coating is compliant with the requirements regarding cytocompatibility stated in the ISO (International Organization for Standardization) 10993 guidelines and an extended release of PMB over a period of at least 42 days was detected. In conclusion, this study serves as a foundation for peptide-releasing hydrogel formulas in the prevention of VAP.


Assuntos
Antibacterianos , Biofilmes , Hidrogéis , Pneumonia Associada à Ventilação Mecânica , Polimixina B , Pseudomonas aeruginosa , Pneumonia Associada à Ventilação Mecânica/prevenção & controle , Polimixina B/farmacologia , Polimixina B/química , Pseudomonas aeruginosa/efeitos dos fármacos , Antibacterianos/farmacologia , Antibacterianos/química , Humanos , Hidrogéis/química , Biofilmes/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Aderência Bacteriana/efeitos dos fármacos , Peptídeos/química , Peptídeos/farmacologia
11.
Microb Pathog ; 196: 106998, 2024 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-39384023

RESUMO

Pseudomonas aeruginosa (P. aeruginosa), a Gram-negative opportunistic pathogen, poses significant treatment challenges due to its antibiotic resistance and biofilm formation. This study investigates the anti-bacterial and anti-biofilm activities of chemically synthesized selenium nanoparticles (SeNPs) against P. aeruginosa. SeNPs were synthesized using ascorbic acid as a reducing agent and characterized. Biofilm formation was quantified using a modified microtiter plate method, and the anti-biofilm efficacy of SeNPs was evaluated using confocal microscopy and SEM. The P. aeruginosa isolates exhibited high resistance to piperacillin-tazobactam (60 %) and ceftazidime (59 %). SeNPs demonstrated a round shape with a diameter of 15-18 nm. UV-Vis spectra showed a peak at 275 nm, and XRD analysis revealed crystalline peaks corresponding to selenium. The FTIR spectra confirmed the presence of various functional groups. SeNPs significantly reduced biofilm formation in a dose-dependent manner, with MIC50 and MIC90 values of 60 µg/mL and 80 µg/mL, respectively. Confocal microscopy and SEM analysis showed a notable decrease in biofilm thickness and bacterial adherence post-SeNPs treatment. These findings suggest that SeNPs could be a promising alternative or adjunctive treatment option for combating antibiotic-resistant P. aeruginosa infections. Further research is warranted to explore the clinical applications of SeNPs in treating biofilm-associated infections.


Assuntos
Antibacterianos , Biofilmes , Testes de Sensibilidade Microbiana , Nanopartículas , Pseudomonas aeruginosa , Selênio , Biofilmes/efeitos dos fármacos , Pseudomonas aeruginosa/efeitos dos fármacos , Selênio/farmacologia , Selênio/química , Antibacterianos/farmacologia , Antibacterianos/química , Nanopartículas/química , Microscopia Confocal , Espectroscopia de Infravermelho com Transformada de Fourier , Aderência Bacteriana/efeitos dos fármacos
12.
BMC Microbiol ; 24(1): 387, 2024 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-39363349

RESUMO

Salmonella forms biofilms, and persist on food contact surfaces. Once a biofilm is formed cleaning and sanitation protocols may be inadequate for effective removal. This study evaluated attachment characteristics, surface properties, and structure of Salmonella biofilms on food contact surfaces commonly used in the tree-fruit industry. Multi-strain Salmonella biofilms were grown in a Centers for Disease Control and Prevention (CDC) biofilm reactor at 22 ± 2 °C and sampling was conducted at 2, 24 and 96-h. After each incubation period, coupons weregently rinsed and the remaining cells enumerated. Biofilms were analyzed with Laser Scanning Confocal Microscopy (LSCM). Hydrophobicity was evaluated by measuring the contact angles of reference liquids method using a drop tensiometer instrument. Material type and biofilm age significantly influenced attachment and biofilm hydrophobicity (P < 0.05). The strength of attachment, across all time points, was highest on nylon followed by wood and high-density polyethylene. The highest contact angle measurements were observed after 96-h of biofilm formation for each material. All the results and observations from this study contribute to a better understanding of the attachment and hydrophobicity characteristics of Salmonella and might help producers make informed decisions when selecting containers for harvesting and storing in order to minimize biofilm formation and potential for cross-contamination.


Assuntos
Aderência Bacteriana , Biofilmes , Microbiologia de Alimentos , Interações Hidrofóbicas e Hidrofílicas , Salmonella , Biofilmes/crescimento & desenvolvimento , Salmonella/fisiologia , Salmonella/crescimento & desenvolvimento , Propriedades de Superfície , Microscopia Confocal , Contaminação de Alimentos
13.
Elife ; 132024 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-39352247

RESUMO

The Gram-negative bacterium Myxococcus xanthus glides on solid surfaces. Dynamic bacterial focal adhesion complexes (bFACs) convert proton motive force from the inner membrane into mechanical propulsion on the cell surface. It is unclear how the mechanical force transmits across the rigid peptidoglycan (PG) cell wall. Here, we show that AgmT, a highly abundant lytic PG transglycosylase homologous to Escherichia coli MltG, couples bFACs to PG. Coprecipitation assay and single-particle microscopy reveal that the gliding motors fail to connect to PG and thus are unable to assemble into bFACs in the absence of an active AgmT. Heterologous expression of E. coli MltG restores the connection between PG and bFACs and thus rescues gliding motility in the M. xanthus cells that lack AgmT. Our results indicate that bFACs anchor to AgmT-modified PG to transmit mechanical force across the PG cell wall.


Assuntos
Parede Celular , Glicosiltransferases , Myxococcus xanthus , Peptidoglicano , Peptidoglicano/metabolismo , Parede Celular/metabolismo , Myxococcus xanthus/genética , Myxococcus xanthus/fisiologia , Myxococcus xanthus/metabolismo , Myxococcus xanthus/enzimologia , Glicosiltransferases/metabolismo , Glicosiltransferases/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Adesões Focais/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Aderência Bacteriana
14.
Antonie Van Leeuwenhoek ; 118(1): 14, 2024 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-39361215

RESUMO

Mammary pathogenic Escherichia coli (MPEC) causes mastitis, which results in substantial economic losses to the dairy industry. A high percentage of Escherichia coli isolated from cows with clinical mastitis harbor adhesin genes, such as fimH. However, it is unclear whether these adhesins are important in the adhesion of MPEC to bovine mammary epithelial cells (BMECs). Therefore, we investigated the effect of adhesins (EcpD, FdeC, and FimH) in MPEC on adherence to the bovine mammary epithelium using cultured BMECs. For this purpose, we used wild-type MPEC as well as single- and double-mutants of fimH, ecpD, and fdeC, and performed adhesion assays with BMECs. First, BMECs were cultured in the presence of lactogenic hormones to induce milk component production and tight junction formation. The bacterial count of the wild-type strain that adhered to the BMECs increased in a dose-dependent manner. In deletion mutant strains, the ΔfimH strain showed lower adhesion (P < 0.05), whereas the adhesion ratio of the ΔecpD and ΔfdeC strains was not statistically different compared with that of the wild-type strain (P > 0.05). Additionally, the fimH/fdeC double-deletion mutants showed the lowest adhesion to BMECs. In conclusion, FimH is crucial in the adhesion of MPEC to BMECs. Overall, our work identifies FimH or FimH/FdeC as interesting targets for future drugs or vaccines to improve the treatment, prevention or chronicity of mastitis induced by MPEC.


Assuntos
Adesinas de Escherichia coli , Aderência Bacteriana , Células Epiteliais , Escherichia coli , Glândulas Mamárias Animais , Animais , Bovinos , Células Epiteliais/microbiologia , Escherichia coli/genética , Adesinas de Escherichia coli/genética , Adesinas de Escherichia coli/metabolismo , Feminino , Glândulas Mamárias Animais/microbiologia , Mastite Bovina/microbiologia , Infecções por Escherichia coli/microbiologia , Proteínas de Fímbrias/genética , Proteínas de Fímbrias/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo
15.
NPJ Biofilms Microbiomes ; 10(1): 98, 2024 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-39358392

RESUMO

Extracellular polysaccharides are crucial components for biofilm development. Although Bacillus subtilis is one of the most characterized Gram-positive biofilm model system, the structure-function of its exopolysaccharide, EpsA-O, remains to be elucidated. By combining chemical analysis, NMR spectroscopy, rheology, and molecular modeling, high-resolution data of EpsA-O structure from atom to supramolecular scale was obtained. The repeating unit is composed of the trisaccharide backbone [→3)-ß-D-QuipNAc4NAc-(1→3)-ß-D-GalpNAc-(1→3)-α-D-GlcpNAc-(1]n, and the side chain ß-D-Galp(3,4-S-Pyr)-(1→6)-ß-D-Galp(3,4-S-Pyr)-(1→6)-α-D-Galp-(1→ linked to C4 of GalNAc. Close agreement between the primary structure and rheological behavior allowed us to model EpsA-O macromolecular and supramolecular solution structure, which can span the intercellular space forming a gel that leads to a complex 3D biofilm network as corroborated by a mutant strain with impaired ability to produce EpsA-O. This is a comprehensive structure-function investigation of the essential biofilm adhesive exopolysaccharide that will serve as a useful guide for future studies in biofilm architecture formation.


Assuntos
Bacillus subtilis , Biofilmes , Espectroscopia de Ressonância Magnética , Polissacarídeos Bacterianos , Biofilmes/crescimento & desenvolvimento , Polissacarídeos Bacterianos/química , Polissacarídeos Bacterianos/metabolismo , Bacillus subtilis/química , Reologia , Modelos Moleculares , Aderência Bacteriana , Sequência de Carboidratos
16.
BMC Genomics ; 25(1): 935, 2024 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-39375592

RESUMO

Listeria monocytogenes is an important foodborne pathogen known for causing listeriosis. To gain insights into the pathogenicity, genetic characterization, and evolution of various Listeria species, in vitro cell adhesion and invasion ability assays and whole-genome sequencing were performed using four Listeria strains isolated from livestock and poultry slaughterhouses. The four Listeria strains exhibited adhesion and invasion abilities in Caco-2 and RAW264.7 cells. Pathogenic Liv1-1 and Lm2-20 had higher adhesion ability, but non-pathogenic Lin4-99 was more invasive than Lm2-20 (p < 0.05). Genetic characterization revealed the presence of a single chromosome without plasmid across four strains with similar whole-genome sizes and G + C% content. Analysis of key pathogenic genes underscored the presence of multiple virulence genes among the four Listeria strains. In contrast, non-pathogenic Listeria lacked LIPI-1, LIPI-2, and LIPI-3 genes, which could possibly be the cause of their non-pathogenicity despite their in vitro cell adhesion and invasion abilities. Thus, genetic determinants of Listeria do not necessarily predict cell adhesion and/or invasive ability in vitro. This study presents a comprehensive comparative genome-wide analysis of four Listeria strains, offering invaluable insights into the pathogenesis of the Listeria genus.


Assuntos
Genoma Bacteriano , Listeria , Fatores de Virulência , Sequenciamento Completo do Genoma , Listeria/genética , Listeria/patogenicidade , Animais , Camundongos , Humanos , Fatores de Virulência/genética , Células CACO-2 , Genômica/métodos , Células RAW 264.7 , Aderência Bacteriana/genética , Virulência/genética , Listeriose/microbiologia , Listeriose/veterinária
17.
World J Microbiol Biotechnol ; 40(11): 367, 2024 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-39455472

RESUMO

Flagella is a well-known bacterial structure crucial for motility, which also plays pivotal roles in pathogenesis. Arcobacter butzleri, an enteropathogen, possesses a distinctive polar flagellum whose functional aspects remain largely unexplored. Upon investigating the factors influencing A. butzleri motility, we uncovered that environmental conditions like temperature, oxygen levels, and nutrient availability play a significant role. Furthermore, compounds that are found in human gut, such as short-chain fatty acids, mucins and bile salts, have a role in modulating the motility, and in turn, the pathogenicity of A. butzleri. Further investigation demonstrated that A. butzleri ΔflaA mutant showed a reduction in motility with a close to null average velocity, as well as a reduction on biofilm formation. In addition, compared with the wild-type, the ΔflaA mutant showed a decreased ability to invade Caco-2 cells and to adhere to mucins. Taken together, our findings support the role of environmental conditions and gut host associated compounds influencing key physiological aspects of the gastrointestinal pathogen A. butzleri, such as motility, and support the role of the flagellum on bacterial virulence.


Assuntos
Arcobacter , Aderência Bacteriana , Biofilmes , Flagelos , Mucinas , Flagelos/genética , Flagelos/fisiologia , Flagelos/metabolismo , Arcobacter/genética , Arcobacter/patogenicidade , Arcobacter/metabolismo , Humanos , Virulência , Células CACO-2 , Biofilmes/crescimento & desenvolvimento , Mucinas/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Ácidos e Sais Biliares/metabolismo , Ácidos e Sais Biliares/farmacologia , Ácidos Graxos Voláteis/metabolismo
18.
J Cell Mol Med ; 28(20): e70182, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39462261

RESUMO

This study explores and verifies potential molecular targets through which KRAS mutations regulate the colonization of Fusobacterium nucleatum (FN) in colorectal cancer (CRC). This study combined multiple bioinformatics methods and biological assays. Through The Cancer Genome Atlas, Gene Expression Omnibus, Human Protein Atlas, immunohistochemistry, and co-culture assays, we further confirmed the differential expression of SERTAD4 in CRC. We delved deeper into examining how expression of SERTAD4 is linked with immune cell infiltration and the enrichment of potential pathways. Lastly, through bacterial phenotypic assays, we validated the function of SERTAD4. As a molecule associated with KRAS mutations and FN infection, the expression levels of SERTAD4 were downregulated in CRC. The diagnostic efficacy of SERTAD4 for CRC is not inferior to that of CEA. Low expression of SERTAD4 is associated with poorer overall survival in CRC. Correlation analysis found that increased expression of SERTAD4 is associated with various immune cell infiltrations and immune checkpoint genes. Finally, bacterial adhesion and invasion assays verify that SERTAD4 inhibits the adhesion and invasion abilities of FN in CRC. This study demonstrates that SERTAD4 exerts a protective role in CRC by inhibiting the colonization of FN.


Assuntos
Neoplasias Colorretais , Fusobacterium nucleatum , Regulação Neoplásica da Expressão Gênica , Mutação , Proteínas Proto-Oncogênicas p21(ras) , Neoplasias Colorretais/microbiologia , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Fusobacterium nucleatum/genética , Fusobacterium nucleatum/patogenicidade , Humanos , Mutação/genética , Proteínas Proto-Oncogênicas p21(ras)/genética , Proteínas Proto-Oncogênicas p21(ras)/metabolismo , Linhagem Celular Tumoral , Regulação para Baixo/genética , Infecções por Fusobacterium/microbiologia , Infecções por Fusobacterium/complicações , Infecções por Fusobacterium/genética , Aderência Bacteriana , Prognóstico
19.
ACS Appl Mater Interfaces ; 16(43): 58226-58240, 2024 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-39420634

RESUMO

In medical infections such as blood sepsis and in food quality control, fast and accurate bacteria analysis is required. Using magnetic nanoparticles (MNPs) for bacterial capture and concentration is very promising for rapid analysis. When MNPs are functionalized with the proper surface chemistry, they have the ability to bind to bacteria and aid in the removal and concentration of bacteria from a sample for further analysis. This study introduces a novel approach for bacterial concentration using polydopamine (pDA), a highly adhesive polymer often purported to create antibacterial and antibiofouling coatings on medical devices. Although pDA has been generally studied for its ability to coat surfaces and reduce biofilm growth, we have found that when coated on magnetic nanoclusters (MNCs), more specifically iron oxide nanoclusters, it effectively binds to and can remove from suspension some types of bacteria. This study investigated the binding of pDA-coated MNCs (pDA-MNCs) to various Gram-negative and Gram-positive bacteria, including Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, and several E. coli strains. MNCs were successfully coated with pDA, and these functionalized MNCs bound a wide variety of bacterial strains. The efficiency of removing bacteria from a suspension can range from 0.99 for S. aureus to 0.01 for an E. coli strain. Such strong capture and differential capture have important applications in collecting bacteria from dilute samples found in medical diagnostics, food and water quality monitoring, and other industries.


Assuntos
Indóis , Nanopartículas de Magnetita , Polímeros , Indóis/química , Polímeros/química , Nanopartículas de Magnetita/química , Aderência Bacteriana/efeitos dos fármacos , Escherichia coli , Staphylococcus epidermidis , Materiais Revestidos Biocompatíveis/química , Materiais Revestidos Biocompatíveis/farmacologia
20.
BMC Oral Health ; 24(1): 1278, 2024 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-39449053

RESUMO

BACKGROUND: Dental restorative materials are recognized as artificial niches that facilitate the adherence and accumulation of oral microorganisms. To mitigate oral diseases and extend the lifespan of restorations, it is advantageous to use dental materials that exhibit low susceptibility to bacterial adhesion. OBJECTIVE: To evaluate and compare bacterial adhesion on two bioactive restorative materials, a glass hybrid restorative, and an alkasite with a nanohybrid resin composite as a positive control. The secondary objectives were to compare the surface roughness (SR) of the materials and determine the correlation between the bacterial adhesion and the SR. MATERIALS AND METHODS: The samples consisted of 33 polished discs of each material: Group A: Tetric® N-Ceram (nanohybrid resin composite), Group B: Equia Forte™ HT Fil (glass hybrid restorative) and Group C: Cention N® (alkasite). Streptococcus mutans cultures were inoculated and after 24-hours of incubation, bacterial adhesion was measured by measuring optical density (OD) and number of colony forming units (CFUs). After 96-hours incubation, the bacterial cell count was determined using scanning electron microscopy (SEM). SR was assessed using surface profilometer. RESULTS: Alkasite had significantly lower OD and CFUs (p < 0.001 and p = 0.015 respectively). According to the SEM analysis, the glass hybrid restorative had lower mean bacterial cell count with no significant difference between the groups. The nanohybrid composite had the smoothest surface that was significantly lower than the alkasite and glass hybrid restorative (p = 0.002). None of the groups demonstrated a correlation between bacterial adhesion and SR. CONCLUSION: Alkasite impedes bacterial adhesion better than the glass hybrid restorative and nanohybrid composite, while smoother surfaces are achieved with the nanohybrid composite.


Assuntos
Aderência Bacteriana , Resinas Compostas , Microscopia Eletrônica de Varredura , Streptococcus mutans , Propriedades de Superfície , Aderência Bacteriana/efeitos dos fármacos , Streptococcus mutans/efeitos dos fármacos , Resinas Compostas/química , Técnicas In Vitro , Cimentos Dentários , Humanos , Teste de Materiais , Cimentos de Resina
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