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1.
Biochemistry ; 63(10): 1257-1269, 2024 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-38683758

RESUMO

Interactions between SJGAP (skipjack tuna GAPDH-related antimicrobial peptide) and four analogs thereof with model bacterial membranes were studied using Fourier-transform infrared spectroscopy (FTIR) and molecular dynamics (MD) simulations. MD trajectory analyses showed that the N-terminal segment of the peptide analogs has many contacts with the polar heads of membrane phospholipids, while the central α helix interacts strongly with the hydrophobic core of the membranes. The peptides also had a marked influence on the wave numbers associated with the phase transition of phospholipids organized as liposomes in both the interface and aliphatic chain regions of the infrared spectra, supporting the interactions observed in the MD trajectories. In addition, interesting links were found between peptide interactions with the aliphatic chains of membrane phospholipids, as determined by FTIR and from the MD trajectories, and the membrane permeabilization capacity of these peptide analogs, as previously demonstrated. To summarize, the combined experimental and computational efforts have provided insights into crucial aspects of the interactions between the investigated peptides and bacterial membranes. This work thus makes an original contribution to our understanding of the molecular interactions underlying the antimicrobial activity of these GAPDH-related antimicrobial peptides from Scombridae.


Assuntos
Simulação de Dinâmica Molecular , Animais , Peptídeos Antimicrobianos/química , Peptídeos Antimicrobianos/farmacologia , Peptídeos Antimicrobianos/metabolismo , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Proteínas de Peixes/química , Proteínas de Peixes/metabolismo , Proteínas de Peixes/farmacologia , Espectroscopia de Infravermelho com Transformada de Fourier , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Gliceraldeído-3-Fosfato Desidrogenases/química , Sequência de Aminoácidos
2.
Int J Mol Sci ; 25(3)2024 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-38338877

RESUMO

Multidrug-resistant Clostridium perfringens infections are a major threat to the poultry industry. Effective alternatives to antibiotics are urgently needed to prevent these infections and limit the spread of multidrug-resistant bacteria. The aim of the study was to produce by chemical synthesis a set of enterocins of different subgroups of class II bacteriocins and to compare their spectrum of inhibitory activity, either alone or in combination, against a panel of twenty C. perfringens isolates. Enterocins A, P, SEK4 (class IIa bacteriocins), B (unsubgrouped class II bacteriocin), and L50 (class IId leaderless bacteriocin) were produced by microwave-assisted solid-phase peptide synthesis. Their antimicrobial activity was determined by agar well diffusion and microtitration methods against twenty C. perfringens isolates and against other pathogens. The FICINDEX of different combinations of the selected enterocins was calculated in order to identify combinations with synergistic effects. The results showed that synthetic analogs of L50A and L50B were the most active against C. perfringens. These peptides also showed the broadest spectrum of activity when tested against other non-clostridial indicator strains, including Listeria monocytogenes, methicillin-resistant Staphylococcus aureus, Streptococcus suis, Streptococcus pyogenes, Enterococcus cecorum, Enterococcus faecalis, as well as Gram-negative bacteria (Campylobacter coli and Pseudomonas aeruginosa), among others. The selected synthetic enterocins were combined on the basis of their different mechanisms of action, and all combinations tested showed synergy or partial synergy against C. perfringens. In conclusion, because of their high activity against C. perfringens and other pathogens, the use of synthetic enterocins alone or as a consortium can be a good alternative to the use of antibiotics in the poultry sector.


Assuntos
Bacteriocinas , Staphylococcus aureus Resistente à Meticilina , Clostridium perfringens , Bacteriocinas/farmacologia , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana , Hidrocarbonetos Aromáticos com Pontes
3.
Food Microbiol ; 101: 103877, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34579845

RESUMO

Salmonella enterica subsp. enterica is one of the leading causes of human foodborne infections and several outbreaks are now associated with the consumption of fresh fruit and vegetables. This study aims at evaluating whether Salmonella virulence can be linked to an enhanced ability to survive successive digestive environments. Thirteen S. enterica strains were selected according to high and low virulence phenotypes. Lettuce inoculated separately with each S. enterica strain was used as food matrix in the TNO gastrointestinal model (TIM-1) of the human upper gastrointestinal tract. During the passage in the stomach, counts determined using PMA-qPCR were 2-5 logs higher than the cultivable counts for all strains indicating the presence of viable but non-cultivable cells. Bacterial growth was observed in the duodenum compartment after 180 min for all but one strain and growth continued into the ileal compartment. After passage through the simulated gastrointestinal tract, both virulent and avirulent S. enterica strains survived but high virulence strains had a significantly (p = 0.004) better average survival rate (1003 %-3753 %) than low virulence strains (from 25 % to 3730%). The survival rates of S. enterica strains could be linked to the presence of genes associated with acid and bile resistance and their predicted products. The presence of single nucleotide polymorphisms may also impact the function of virulence associated genes and play a role in the resulting phenotype. These data provide an understanding of the relationship between measured virulence potential and survival of S. enterica during dynamic simulated gastrointestinal transit.


Assuntos
Trato Gastrointestinal/microbiologia , Salmonella/patogenicidade , Virulência , Humanos , Modelos Biológicos
4.
Appl Environ Microbiol ; 87(23): e0139621, 2021 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-34550762

RESUMO

The adhesion of noroviruses to strawberry, turkey slices, ham, and cheddar cheese was studied using murine norovirus 1 (MNV-1) as a surrogate for human norovirus (NoV). Based on plaque assay, the recovery and adhesion of MNV-1 depended on the food type (turkey versus strawberry), pH of the initial suspension buffer (pH 4 versus pH 7), and food fat composition (C8 versus C18). Recovery of infectious particles from turkey was 68% compared to 9.4% from strawberry. On turkey, adhesion of MNV-1 was lower at pH 7 (pH of fecal matter), and virus particles adhered to this pH were recovered more easily (33,875 PFU) than at pH 4 (pH of vomitus). The presence of fat and the composition of fatty acids seemed to increase MNV-1 recovery and adherent viral particles recovered but did not affect adhesion (68% on fat-free turkey and regular turkey). Adherent MNV-1 particles recovered from stainless steel coated with saturated fatty acid (C8, C14, C18) increased significantly with chain length (P < 0.05), but adhesion did not seem to change. Using liquid droplet contact angle to measure surface energy, it was deduced that hydrophobic interactions contribute considerably to the adhesion of MNV-1 to stainless steel, polyvinyl chloride, and high-density polyethylene. IMPORTANCE Ready-to-eat (RTE) foods are major vehicles of transmission of foodborne viral pathogens, including NoV. The high incidence of gastroenteritis caused by viruses is due largely to their persistence in the environment and adhesion to different kinds of surfaces in the food industry, including the foods themselves. Compared with bacteria, adhesion of viruses to surfaces is poorly understood. Better knowledge of the physicochemical parameters involved in the adhesion of NoV to ready-to-eat foods is essential to devising effective strategies for reducing the persistence and, thus, the transmission of this virus.


Assuntos
Fast Foods/virologia , Contaminação de Alimentos/análise , Norovirus , Queijo/virologia , Frutas/virologia , Interações Hidrofóbicas e Hidrofílicas , Carne/virologia , Aço Inoxidável
5.
Arch Microbiol ; 203(1): 205-217, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32803347

RESUMO

Here, we have analysed and explored the genome sequences of three newly isolated bacteria that were recently characterised for their probiotic activities and ability to produce bacteriocins. These strains, isolated from faeces of animals living in captivity at the zoological garden of Lille (France), are Escherichia coli ICVB443, Enterococcus faecalis ICVB501 and Pediococcus pentosaceus ICVB491. Their genomes have been analysed and compared to those of their pathogenic or probiotic counterparts. The genome analyses of E. coli ICVB443 and Ent. faecalis ICVB501 displayed similarities to those of probiotics E. coli 1917 Nissle, and Ent. faecalis Symbioflor 1, respectively. Furthermore, E. coli ICVB443 shares at least 89 genes with the enteroaggregative E. coli 55989 (EAEC), and Ent. faecalis ICVB501 shares at least 315 genes with the pathogenic Ent. faecalis V583 strain. Unlike Ped. pentosaceus ICVB491, which is devoid of virulence genes, E. coli ICVB443 and Ent. faecalis ICVB501 both carry genes encoding virulence factors on their genomes. Of note, the bioinformatics analysis of these two genomes located the bsh gene, which codes for bile salt hydrolase (BSH). The presence of BSH is of major importance, as it can help to increase the viability of these two strains in the gastrointestinal tract (GIT). The genome analysis of Ped. pentosaceus ICVB491 confirmed its GRAS status (Generally Recognised As Safe), as no genomic virulence factor determinant was found.


Assuntos
Bactérias/genética , Bacteriocinas/genética , Fezes/microbiologia , Genoma Bacteriano/genética , Animais , Bactérias/patogenicidade , Simulação por Computador , Enterococcus faecalis/genética , Enterococcus faecalis/patogenicidade , Escherichia coli/genética , Escherichia coli/patogenicidade , Trato Gastrointestinal/microbiologia , Pediococcus pentosaceus/genética , Pediococcus pentosaceus/patogenicidade , Probióticos , Fatores de Virulência/genética
6.
Int J Mol Sci ; 22(10)2021 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-34068193

RESUMO

In this study, we investigate the immunomodulatory effects of a novel antimicrobial peptide, YD1, isolated from Kimchi, in both in vitro and in vivo models. We establish that YD1 exerts its anti-inflammatory effects via up-regulation of the Nrf2 pathway, resulting in the production of HO-1, which suppresses activation of the NF-κB pathway, including the subsequent proinflammatory cytokines IL-1ß, IL-6, and TNF-α. We also found that YD1 robustly suppresses nitric oxide (NO) and prostaglandin E2 (PGE2) production by down-regulating the expression of the upstream genes, iNOS and COX-2, acting as a strong antioxidant. Collectively, YD1 exhibits vigorous anti-inflammatory and antioxidant activity, presenting it as an interesting potential therapeutic agent.


Assuntos
Anti-Inflamatórios/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Heme Oxigenase-1/metabolismo , Inflamação/prevenção & controle , Proteínas de Membrana/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Proteínas Citotóxicas Formadoras de Poros/farmacologia , Animais , Citocinas/metabolismo , Edema/induzido quimicamente , Edema/metabolismo , Edema/patologia , Edema/prevenção & controle , Heme Oxigenase-1/genética , Inflamação/induzido quimicamente , Inflamação/metabolismo , Inflamação/patologia , Lipopolissacarídeos/toxicidade , Proteínas de Membrana/genética , Camundongos , Fator 88 de Diferenciação Mieloide/antagonistas & inibidores , Fator 88 de Diferenciação Mieloide/genética , Fator 88 de Diferenciação Mieloide/metabolismo , Fator 2 Relacionado a NF-E2/genética , NF-kappa B/antagonistas & inibidores , NF-kappa B/genética , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Receptor 4 Toll-Like/antagonistas & inibidores , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo
7.
Environ Microbiol ; 22(7): 2907-2920, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32363677

RESUMO

In livestock production, antibiotics are used to promote animal growth, control infections and thereby increase profitability. This practice has led to the emergence of multiresistant bacteria such as Salmonella, of which some serovars are disseminated in the environment. The objective of this study is to evaluate microcin J25 as an inhibitor of Salmonella enterica serovars of various origins including human, livestock and food. Among the 116 isolates tested, 37 (31.8%) were found resistant to at least one antibiotic, and 28 were multiresistant with 19 expressing the penta-resistant phenotype ACSSuT. Microcin J25 inhibited all isolates, with minimal inhibitory concentration values ranging from 0.06 µg/ml (28.4 nM) to 400 µg/ml (189 µM). Interestingly, no cross-resistance was found between microcin J25 and antibiotics. Multiple sequence alignments of genes encoding for the different proteins involved in the recognition and transport of microcin J25 showed that only ferric-hydroxamate uptake is an essential determinant for susceptibility of S. enterica to microcin J25. Examination of Salmonella strains exposed to microcin J25 by transmission electronic microscopy showed for the first-time involvement of a pore formation mechanism. Microcin J25 was a strong inhibitor of several multiresistant isolates of Salmonella and may have a great potential as an alternative to antibiotics.


Assuntos
Bacteriocinas/farmacologia , Salmonella enterica/genética , Animais , Antibacterianos/farmacologia , Genômica , Humanos , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Transmissão , Fenômica , Salmonella enterica/efeitos dos fármacos , Salmonella enterica/ultraestrutura
8.
FASEB J ; 33(4): 4921-4935, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30596521

RESUMO

Given the growing evidence that gut dysfunction, including changes in gut microbiota composition, plays a critical role in the development of inflammation and metabolic diseases, the identification of novel probiotic bacteria with immunometabolic properties has recently attracted more attention. Herein, bacterial strains were first isolated from dairy products and human feces and then screened in vitro for their immunomodulatory activity. Five selected strains were further analyzed in vivo, using a mouse model of diet-induced obesity. C57BL/6 mice were fed a high-fat high-sucrose diet, in combination with 1 of 3 Lactobacillus strains (Lb38, L. plantarum; L79, L. paracasei/casei; Lb102, L. rhamnosus) or Bifidobacterium strains (Bf26, Bf141, 2 different strains of B. animalis ssp. lactis species) administered for 8 wk at 109 colony-forming units/d. Whereas 3 strains showed only modest (Lb38, Bf26) or no (L79) effects, Lb102 and Bf141 reduced diet-induced obesity, visceral fat accretion, and inflammation, concomitant with improvement of glucose tolerance and insulin sensitivity. Further analysis revealed that Lb102 and Bf141 enhanced intestinal integrity markers in association with selective changes in gut microbiota composition. We have thus identified 2 new potential probiotic bacterial strains with immunometabolic properties to alleviate obesity development and associated metabolic disturbances.-Le Barz, M., Daniel, N., Varin, T. V., Naimi, S., Demers-Mathieu, V., Pilon, G., Audy, J., Laurin, E., Roy, D., Urdaci, M. C., St-Gelais, D., Fliss, I, Marette, A. In vivo screening of multiple bacterial strains identifies Lactobacillus rhamnosus Lb102 and Bifidobacterium animalis ssp. lactis Bf141 as probiotics that improve metabolic disorders in a mouse model of obesity.


Assuntos
Bifidobacterium animalis/fisiologia , Lacticaseibacillus rhamnosus/fisiologia , Obesidade/dietoterapia , Obesidade/microbiologia , Probióticos/uso terapêutico , Tecido Adiposo/metabolismo , Animais , Quimiocinas/metabolismo , Citocinas/metabolismo , Modelos Animais de Doenças , Ácidos Graxos não Esterificados/metabolismo , Microbioma Gastrointestinal/fisiologia , Inflamação/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Obesidade/metabolismo , RNA Ribossômico 16S/genética
9.
Mar Drugs ; 17(7)2019 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-31336895

RESUMO

The Atlantic mackerel, Scomber scombrus, is one of the most fished species in the world, but it is still largely used for low-value products, such as bait; mainly for crustacean fishery. This resource could be transformed into products of high value and may offer new opportunities for the discovery of bioactive molecules. Mackerel hydrolysate was investigated to discover antibacterial peptides with biotechnological potential. The proteolytic process generated a hydrolysate composed of 96% proteinaceous compounds with molecular weight lower than 7 kDa. From the whole hydrolysate, antibacterial activity was detected against both Gram-negative and Gram-positive bacteria. After solid phase extraction, purification of the active fraction led to the identification of 4 peptide sequences by mass spectrometry. The peptide sequence N-KVEIVAINDPFIDL-C, called Atlantic Mackerel GAPDH-related peptide (AMGAP), was selected for chemical synthesis to confirm the antibacterial activity and to evaluate its stability through in vitro digestibility. Minimal inhibitory concentrations of AMGAP revealed that Listeria strains were the most sensitive, suggesting potential as food-preservative to prevent bacterial growth. In addition, in vitro digestibility experiments found rapid (after 20 min) and early digestibility (stomach). This study highlights the biotechnological potential of mackerel hydrolysate due to the presence of the antibacterial AMGAP peptide.


Assuntos
Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/farmacologia , Conservantes de Alimentos/farmacologia , Perciformes , Hidrolisados de Proteína/farmacologia , Animais , Antibacterianos/química , Antibacterianos/isolamento & purificação , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/isolamento & purificação , Biotecnologia/métodos , Conservantes de Alimentos/química , Conservantes de Alimentos/isolamento & purificação , Mucosa Gástrica/metabolismo , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Hidrolisados de Proteína/química , Hidrolisados de Proteína/isolamento & purificação , Estabilidade Proteica , Proteólise , Suínos
10.
Acta Vet Hung ; 67(4): 477-488, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31842593

RESUMO

The spread of antimicrobial-resistant bacteria in wildlife must be viewed as a major concern with serious implications for human and animal health. Escherichia coli and enterococcal isolates were recovered from faecal samples of 49 wild rabbits (Oryctolagus cuniculus) on specific media and were characterised using biochemical and molecular tests. For all isolates, antimicrobial susceptibility testing was performed, and resistance genes were detected by PCR. Molecular typing of isolates was carried out by pulsed-field gel-electrophoresis, and E. coli strains were also tested for the presence of intimin (eae) gene characteristic of rabbit enteropathogenic E. coli. A total of 34 E. coli and 36 enterococci [E. hirae (52.8%) and E. faecalis (47.2%)] were obtained. For E. coli, resistance to tetracycline (94%), streptomycin (62%), ciprofloxacin (47%), trimethoprim-sulphamethoxazole (35%) and chloramphenicol (6%) was observed. Resistance to third-generation cephalosporins was detected in one E. coli strain that carried the blaCMY-2 and blaTEM-1 genes. Class 1 integrons were detected in eight isolates. For enterococci, resistance to tetracycline (63.9%), erythromycin (30.5%), streptomycin (18.2%), and chloramphenicol (5.5%) was detected. The tet(M)+tet(L), erm(B) and ant (6)-Ia genes were identified in thirteen, seven and three resistant Enterococcus strains, respectively. Molecular typing showed a high diversity among our strains. Wild rabbits could represent a reservoir of E. coli, and enterococci carrying antimicrobial resistance genes and E. coli additionally carrying the eae gene of enteropathogenic pathotypes could both contaminate the environment. our finding seems to represent the first report of eae-positive E. coli in wild rabbits.


Assuntos
Adesinas Bacterianas/genética , Antibacterianos/farmacologia , Farmacorresistência Bacteriana/genética , Enterococcus/efeitos dos fármacos , Infecções por Escherichia coli/veterinária , Proteínas de Escherichia coli/genética , Escherichia coli/efeitos dos fármacos , Infecções por Bactérias Gram-Positivas/veterinária , Adesinas Bacterianas/metabolismo , Animais , Enterococcus/genética , Enterococcus/patogenicidade , Escherichia coli/genética , Escherichia coli/patogenicidade , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli/metabolismo , Infecções por Bactérias Gram-Positivas/epidemiologia , Infecções por Bactérias Gram-Positivas/microbiologia , Prevalência , Coelhos , Tunísia/epidemiologia , Virulência/genética
11.
Arch Microbiol ; 200(5): 771-782, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29417164

RESUMO

The aim of this study was to isolate and investigate the bacteriocinogenic and probiotic potential of new Gram-negative isolates. Of 22 bacterial isolates from pig intestine and chicken crops, ten isolates had demonstrated a good activity, and the most potent five strains were identified as four E. coli and one as Proteus sp. No virulence factors were detected for E. coli strains isolated from pig intestine. The semi-purified microcins proved to be resistant to temperature and pH variation, but sensitive to proteolytic enzymes. Of particular interest, strain E. coli P2C was the most potent, free of virulence genes and sensitive to tested antibiotics. Purification procedure revealed the presence of a single pure peak having a molecular mass of 8733.94 Da and matching microcin V (MccV). The sequence obtained by LC-MS/MS confirmed the presence of MccV. Purified MccV showed a good activity against pathogenic coliforms, especially E. coli O1K1H7 involved in avian colibacillosis. The present study provides evidence that E. coli strains isolated from pig intestine produce microcin-like substances. E. coli P2C is a safe MccV producer that could be a good candidate for its application as novel probiotic strain to protect livestock and enhance growth performance.


Assuntos
Bacteriocinas/isolamento & purificação , Escherichia coli/genética , Animais , Bacteriocinas/química , Bacteriocinas/farmacologia , Galinhas/microbiologia , Cromatografia Líquida de Alta Pressão , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/isolamento & purificação , Trato Gastrointestinal/microbiologia , Tipagem Molecular , Probióticos/análise , Proteus/genética , Proteus/crescimento & desenvolvimento , Proteus/isolamento & purificação , RNA Ribossômico 16S/genética , Sus scrofa/microbiologia
12.
Arch Microbiol ; 199(2): 215-222, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27671474

RESUMO

Staphylococcus aureus and methicillin-resistant S. aureus are major pathogens. The antimicrobial peptides and essential oils (EOs) display narrow- or broad-spectrum activity against bacteria including these strains. A centralized resource, such as a database, designed specifically for anti-S. aureus/anti-methicillin-resistant S. aureus antimicrobial peptides and EOs is therefore needed to facilitate the comprehensive investigation of their structure/activity associations and combinations. The database ANTISTAPHYBASE is created to facilitate access to important information on antimicrobial peptides and essential peptides against methicillin-resistant S. aureus and S. aureus. At the moment, the database contains 596 sequences of antimicrobial peptides produced by diverse organisms and 287 essential oil records. It permits a quick and easy search of peptides based on their activity as well as their general, physicochemical properties and literature data. These data are very useful to perform further bioinformatic or chemometric analysis and would certainly be useful for the development of new drugs for medical use. The ANTISTAPHYBASE database is freely available at: https://www.antistaphybase.com/ .


Assuntos
Antibacterianos/farmacologia , Bases de Dados de Produtos Farmacêuticos , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Óleos Voláteis/farmacologia , Peptídeos/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Antibacterianos/química , Peptídeos/química , Relação Estrutura-Atividade
13.
Anaerobe ; 47: 173-182, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28578132

RESUMO

In this work, the synergistic effect of electro-activated solutions (EAS) of potassium acetate and potassium citrate, nisin and moderate heat treatment to inactivate C. sporogenes PA 3679 spores was evaluated in green beans puree and whole green beans. Electro-activated solutions (EAS) of potassium acetate and potassium citrate were generated under 400 mA during 60 min. They were characterized by an oxidation-reduction potential (ORP) and pH values ranged from +300 to +1090 mV and 2.8 to 3.67, respectively. Moreover, the EAS were combined with a bacteriocin nisin at concentrations of 250, 500, 750 and 1000 IU/mL and the targeted sporicidal effect was evaluated under moderate heat treatment. The inoculated mixtures were subjected to temperatures of 95, 105 and 115 °C for exposure times of 5, 15 and 30 min. After plate counting, the synergistic effect of the hurdle principle composed of electro-activated solutions, nisin and moderate temperatures was demonstrated. The obtained results showed that the synergistic effect of the used hurdle was able to achieve an inactivation efficacy of 5.9-6.1 log CFU/mL. Furthermore, experiments carried out with whole green beans showed that spore inactivation level was significantly higher and reach 6.5 log CFU/mL. Moreover, spore morphology was examined by transmission electron microscopy and the obtained micrographs showed important damages in all of the treated spores.


Assuntos
Antibacterianos/farmacologia , Clostridium/efeitos dos fármacos , Clostridium/efeitos da radiação , Nisina/farmacologia , Soluções/farmacologia , Esporos Bacterianos/efeitos dos fármacos , Esporos Bacterianos/efeitos da radiação , Contagem de Colônia Microbiana , Microbiologia de Alimentos , Temperatura Alta , Phaseolus/microbiologia , Acetato de Potássio/farmacologia , Citrato de Potássio/farmacologia , Soluções/química
14.
J Microencapsul ; 34(2): 171-184, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28350268

RESUMO

Colon targeting, as a site-specific delivery for oral formulation, remains a major challenge, especially for sensitive bioactive components such as therapeutic forms of phages, live attenuated virus and prebiotics-probiotics association. Synbiotics could be used to protect encapsulated probiotics during the gastrointestinal tract and control their release in the colon. To achieve these goals, effective prebiotics, such as inulin, could be combined with alginate - the most exploited polymer used for probiotic encapsulation - in the form of beads. This work aimed to study the biopharmaceutical behaviour of alginate beads (A) and inulin-alginate beads of different inulin concentrations (5 or 20%) in 2% alginate (AI5, AI20). Beads were loaded with three probiotic strains (Pediococcus acidilactici Ul5, Lactobacillus reuteri and Lactobacillus salivarius). Dissolution of beads was studied by USP4 under conditions simulating the gastrointestinal condition. The survival rates of the bacterial strains were measured by a specific qPCR bacterial count. Mucoadhesiveness of beads was studied by an ex vivo method using intestinal mucosa. To understand the behaviour of each formulation, the ultrastructure of the polymeric network was studied using scanning electron microscopy (SEM). Molecular interactions between alginate and inulin were studied by Fourier transform infra-red spectroscopy (FTIR). Dissolution results suggested that the presence of inulin in beads provided more protection for the tested bacterial strains against the acidic pH. AI5 was the most effective formulation to deliver probiotics to the colon simulation conditions. FTIR and SEM investigations explained the differences in behaviour of each formula. The developed symbiotic form provided a promising matrix for the development of colonic controlled release systems.


Assuntos
Alginatos/farmacologia , Inulina/farmacologia , Probióticos , Simbióticos , Colo , Formas de Dosagem
15.
Biochem Biophys Res Commun ; 473(2): 642-7, 2016 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-27038545

RESUMO

In this study, we first report characterization of collagencin, an antimicrobial peptide identified from fish collagen hydrolysate. The peptide completely inhibited the growth of Staphylococcus aureus at 1.88 mM. Although non-toxic up to 470 µM, collagencin was hemolytic at higher concentrations. The secondary structure of collagencin was mainly composed by ß-sheet and ß-turn as determined by CD measurements and molecular dynamics. The peptide is likely to form ß-sheet structure under hydrophobic environments and interacts with both anionic (phosphatidylglycerol) and zwitterionic (phosphoethanolamine and phosphatidylcholine) lipids as shown with CD spectroscopy and molecular dynamics. The peptide formed several hydrogen bonds with both POPG and POPE lipids and remained at membrane-water interface, suggesting that collagencin antibacterial action follows a carpet mechanism. Collagenous fish wastes could be processed by enzymatic hydrolysis and transformed into products of high value having functional or biological properties. Marine collagens are a promising source of antimicrobial peptides with new implications in food safety and human health.


Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/farmacologia , Colágeno/química , Colágeno/farmacologia , Sequência de Aminoácidos , Animais , Dicroísmo Circular , Peixes/metabolismo , Humanos , Lipídeos de Membrana/metabolismo , Simulação de Dinâmica Molecular , Estrutura Secundária de Proteína , Infecções Estafilocócicas/tratamento farmacológico , Staphylococcus aureus/efeitos dos fármacos
16.
J Nutr ; 146(11): 2206-2215, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27708120

RESUMO

BACKGROUND: MicroRNAs are small, gene-regulatory noncoding RNA species present in large amounts in milk, where they seem to be protected against degradative conditions, presumably because of their association with exosomes. OBJECTIVE: We monitored the relative stability of commercial dairy cow milk microRNAs during digestion and examined their associations with extracellular vesicles (EVs). METHODS: We used a computer-controlled, in vitro, gastrointestinal model TNO intestinal model-1 (TIM-1) and analyzed, by quantitative polymerase chain reaction, the concentration of 2 microRNAs within gastrointestinal tract compartments at different points in time. EVs within TIM-1 digested and nondigested samples were studied by immunoblotting, dynamic light scattering, quantitative polymerase chain reaction, and density measurements. RESULTS: A large quantity of dairy milk Bos taurus microRNA-223 (bta-miR-223) and bta-miR-125b (∼109-1010 copies/300 mL milk) withstood digestion under simulated gastrointestinal tract conditions, with the stomach causing the most important decrease in microRNA amounts. A large quantity of these 2 microRNAs (∼108-109 copies/300 mL milk) was detected in the upper small intestine compartments, which supports their potential bioaccessibility. A protocol optimized for the enrichment of dairy milk exosomes yielded a 100,000 × g pellet fraction that was positive for the exosomal markers tumor susceptibility gene-101 (TSG101), apoptosis-linked gene 2-interacting protein X (ALIX), and heat shock protein 70 (HSP70) and containing bta-miR-223 and bta-miR-125b. This approach, based on successive ultracentrifugation steps, also revealed the existence of ALIX-, HSP70-/low, and TSG101-/low EVs larger than exosomes and 2-6 times more enriched in bta-miR-223 and bta-miR-125b (P < 0.05). CONCLUSIONS: Our findings indicate that commercial dairy cow milk contains numerous microRNAs that can resist digestion and are associated mostly with ALIX-, HSP70-/low, and TSG101-/low EVs. Our results support the existence of interspecies transfer of microRNAs mediated by milk consumption and challenge our current view of exosomes as the sole carriers of milk-derived microRNAs.


Assuntos
Bovinos , Digestão , MicroRNAs/química , MicroRNAs/metabolismo , Leite/química , Animais , Exossomos , Trato Gastrointestinal , Modelos Biológicos , Fatores de Tempo
17.
Microb Ecol ; 72(4): 931-942, 2016 11.
Artigo em Inglês | MEDLINE | ID: mdl-26162534

RESUMO

Pediococcus acidilactici UL5 is a promising probiotic candidate due to its high survival rate under gastric and duodenal conditions and to its ability to produce the antilisterial bacteriocin pediocin PA-1. Its survival, metabolic activity, and impact on Listeria monocytogenes in a continuous stirred tank reactor containing immobilized human intestinal microbiota were studied over a period of 32 days of feeding a nutrient medium simulating ileal chyme. The impact of P. acidilactici UL5 on different bacterial groups of intestinal origin as well as its survival and its impact on L. monocytogenes were quantified using quantitative polymerase chain reaction coupling with propidium monoazide (PMA-qPCR), which was shown to detect and quantify viable bacteria only. P. acidilactici UL5 and its non-pediocin-producing mutant had no effect on the microbiota, but the producing strain induced an increase in the production of acetic and propionic acids. P. acidilactici survived but appeared to be a poor competitor with intestinal microbiota, dropping by 1.3 and 2.8 log10 after 8 h of fermentation to 104 colony-forming units (cfu) mL-1. A 1.64 log but non-significant reduction of Listeria was observed when P. acidilactici UL5 was added at 108 cfu mL-1. P. acidilactici UL5 isolated from the reactor after 3 days was still able to produce the active bacteriocin. These data demonstrate that P. acidilactici UL5 is capable of surviving transit through the ileum without losing its ability to produce pediocin PA-1 but seems to not be enough competitive with the great diversity of organisms found in the ileum.


Assuntos
Microbioma Gastrointestinal/fisiologia , Íleo/microbiologia , Listeria monocytogenes/crescimento & desenvolvimento , Pediocinas/metabolismo , Pediococcus acidilactici/metabolismo , Probióticos/metabolismo , Azidas/farmacologia , Fermentação , Humanos , Propídio/análogos & derivados , Propídio/farmacologia , RNA Ribossômico 16S/genética , Reação em Cadeia da Polimerase em Tempo Real
18.
J Microencapsul ; 33(1): 89-101, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26805512

RESUMO

This work aims to develop an encapsulated oral-synbiotic supplement by studying the effect of adding inulin in alginate beads and observing its ability to protect three probiotic strains: Pediocucus acidilactici, Lactobacillus reuteri and Lactobacillus salivarius. Beads of different inulin concentrations 0%, 5%, 10%, 15% and 20% (w/v) in 2% (w/v) alginate solution were prepared by the extrusion/ionotropic gelation method. Polymer distribution within beads was characterised using confocal laser scanning microscopy. Interactions between alginate and inulin were monitored by Fourier transform infra-red spectroscopy (FTIR). Effect of encapsulation on viability, antimicrobial ability, acid tolerance and bile tolerance of probiotic strains were investigated. Antimicrobial and probiotic properties of bacterial strains were not affected by encapsulation. Bacterial protection against acidity was increased by adding inulin. Beads with 5% w/v inulin were the most effective in bacterial protection against bile-salts. To our knowledge, this work is the first to use such high concentrations of inulin.


Assuntos
Alginatos/química , Inulina/química , Limosilactobacillus reuteri/metabolismo , Prebióticos/microbiologia , Células Imobilizadas/metabolismo , Ácido Glucurônico/química , Ácidos Hexurônicos/química , Limosilactobacillus reuteri/química
19.
Biochem Biophys Res Commun ; 462(3): 195-200, 2015 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-25934151

RESUMO

Proteins from fish by-product sources are valuable source of bioactive peptides and show promise as functional foods ingredients. The objective of the present study was to isolate and characterize antibacterial peptides from protamex hydrolysates of Atlantic mackerel (Scomber scombrus) by-products. Four sequences SIFIQRFTT (P4), RKSGDPLGR (P8.1), AKPGDGAGSGPR (P8.2) and GLPGPLGPAGPK (P11) were identified in peptide fractions separated using RP-HPLC. At 200 µg mL(-1), while peptides P8.1, P8.2 and P11 exhibited partial inhibition, P4 totally inhibited tested Gram-positive (Listeria innocua) and Gram-negative (Escherichia coli) bacterial strains. These results suggest that the protein hydrolysate derived from mackerel by-products could be used as an antimicrobial ingredient in both functional food and nutraceutical applications.


Assuntos
Antibacterianos/isolamento & purificação , Peptídeos Catiônicos Antimicrobianos/isolamento & purificação , Proteínas de Peixes/isolamento & purificação , Perciformes/metabolismo , Sequência de Aminoácidos , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/genética , Peptídeos Catiônicos Antimicrobianos/farmacologia , Escherichia coli/efeitos dos fármacos , Proteínas de Peixes/genética , Proteínas de Peixes/farmacologia , Hidrólise , Listeria/efeitos dos fármacos , Dados de Sequência Molecular , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/isolamento & purificação , Fragmentos de Peptídeos/farmacologia , Perciformes/genética , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
20.
Appl Environ Microbiol ; 81(8): 2950-7, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25681193

RESUMO

Pulsed light is a nonthermal processing technology recognized by the FDA for killing microorganisms on food surfaces, with cumulative fluences up to 12 J cm(-2). In this study, we investigated its efficacy for inactivating murine norovirus 1 (MNV-1) as a human norovirus surrogate in phosphate-buffered saline, hard water, mineral water, turbid water, and sewage treatment effluent and on food contact surfaces, including high-density polyethylene, polyvinyl chloride, and stainless steel, free or in an alginate matrix. The pulsed-light device emitted a broadband spectrum (200 to 1,000 nm) at a fluence of 0.67 J cm(-2) per pulse, with 2% UV at 8 cm beneath the lamp. Reductions in viral infectivity exceeded 3 log10 in less than 3 s (5 pulses; 3.45 J cm(-2)) in clear suspensions and on clean surfaces, even in the presence of alginate, and in 6 s (11 pulses; 7.60 J cm(-2)) on fouled surfaces except for stainless steel (2.6 log10). The presence of protein or bentonite interfered with viral inactivation. Analysis of the morphology, the viral proteins, and the RNA integrity of treated MNV-1 allowed us to elucidate the mechanisms involved in the antiviral activity of pulsed light. Pulsed light appeared to disrupt MNV-1 structure and degrade viral protein and RNA. The results suggest that pulsed-light technology could provide an effective alternative means of inactivating noroviruses in wastewaters, in clear beverages, in drinking water, or on food-handling surfaces in the presence or absence of biofilms.


Assuntos
Descontaminação/métodos , Desinfecção/métodos , Água Potável/virologia , Norovirus/efeitos da radiação , Inativação de Vírus/efeitos da radiação , Águas Residuárias/virologia , Descontaminação/instrumentação , Desinfecção/instrumentação , Norovirus/fisiologia
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