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1.
Food Chem ; 447: 138951, 2024 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-38489883

RESUMO

Biocomplex materials formed by oppositely charged biopolymers (proteins) tend to be sensitive to environmental conditions and may lose part functional properties of original proteins, and one of the approaches to address these weaknesses is protein modification. This study established an electrostatic composite system using succinylated ovalbumin (SOVA) and ε-polylysine (ε-PL) and investigated the impact of varying degrees of succinylation and ε-PL addition on microstructure, environmental responsiveness and functional properties. Molecular docking illustrated that the most favorable binding conformation was that ε-PL binds to OVA groove, which was contributed by the multi­hydrogen bonding and hydrophobic interactions. Transmission electron microscopy observed that SOVA/ε-PL had a compact spherical structure with 100 nm. High-degree succinylation reduced complex sensitivity to heat, ionic strength, and pH changes. ε-PL improved the gel strength and antibacterial properties of SOVA. The study suggests possible uses of SOVA/ε-PL complex as multifunctional protein complex systems in the field of food additives.


Assuntos
Antibacterianos , Polilisina , Polilisina/química , Ovalbumina , Eletricidade Estática , Simulação de Acoplamento Molecular
2.
Microbiol Res ; 275: 127461, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37499310

RESUMO

Owing to the threats that Salmonella poses to public health and the abuse of antimicrobials, bacteriophage therapy against Salmonella is experiencing a resurgence. Although several phages have been reported as safe and efficient for controlling Salmonella, the genetic diversity and relatedness among Salmonella phages remain poorly understood. In this study, whole-genome sequences of 91 Salmonella bacteriophages were obtained from the National Center for Biological Information genome database. Phylogenetic analysis, mosaic structure comparisons, gene content analysis, and orthologue group clustering were performed. Phylogenetic analysis revealed four singletons and two major lineages (I-II), including five subdividing clades, of which Salmonella phages belonging to morphologically distinct families were clustered in the same clade. Chimeric structures (n = 31), holin genes (n = 18), lysin genes (n = 66), DNA packaging genes (n = 55), and DNA metabolism genes (n = 24) were present in these phages. Moreover, phages from different subdivided clusters harboured distinct genes associated with host cell lysis, DNA packaging, and DNA metabolism. Notably, phages belonging to morphologically distinct families shared common orthologue groups. Although several functional modules of phages SS1 and SE16 shared > 99% nucleotide sequence identity with phages SI2 and SI23, the major differences between these phages were the absence and replication of functional modules. The data obtained herein revealed the genetic diversity of Salmonella phages at genomic, structural, and gene content levels. The genetic diversity of Salmonella phages is likely owing to the acquisition, loss, and replication of functional modules.


Assuntos
Bacteriófagos , Fagos de Salmonella , Humanos , Fagos de Salmonella/genética , Filogenia , Genoma Viral , Bacteriófagos/genética , Salmonella/genética , DNA , Variação Genética
3.
Food Res Int ; 165: 112511, 2023 03.
Artigo em Inglês | MEDLINE | ID: mdl-36869511

RESUMO

In this study, the functional properties of ovalbumin (OVA) were improved through dual modification with succinylation (succinylation degrees of 32.1 % [S1], 74.2 % [S2], and 95.2 % [S3]) and ultrasonication (ultrasonication durations of 5 min [U1], 15 min [U2], and 25 min [U3]), and the changes in protein structures were explored. Results showed that as the succinylation degree was increased, the particle size and surface hydrophobicity of S-OVA decreased by the maximum values of 2.2 and 2.4 times, respectively, causing emulsibility and emulsifying stability to increase by 2.7 and 7.3 times, respectively. After ultrasonic treatment, the particle size of succinylated-ultrasonicated OVA (SU-OVA) had decreased by 3.0-5.1 times relative to that of S-OVA. Moreover, the net negative charge of S3U3-OVA had increased to the maximum value of - 35.6 mV. These changes contributed to the further enhancement in functional indicators. The unfolding of the protein structure and the conformational flexibility of SU-OVA were illustrated and compared with those of S-OVA via protein electrophoresis, circular dichroism spectroscopy, intrinsic fluorescence spectroscopy, and scanning electron microscopy. The dually modified OVA emulsion (S3U3-E) presented small droplets (243.33 nm), reduced viscosity, and weakened gelation behavior that were indicative of even distribution, which was visually proven by confocal laser scanning microscopy images. Furthermore, S3U3-E exhibited favorable stability, a particle size that was almost unchanged, and a low polydispersity index (<0.1) over 21 days of storage at 4 °C. The above results demonstrated that succinylation combined with ultrasonic treatment could be an effective dual modification method for enhancing the functional performance of OVA.


Assuntos
Ovalbumina , Microscopia Confocal , Microscopia Eletrônica de Varredura , Tamanho da Partícula , Espectrometria de Fluorescência
4.
Microb Pathog ; 174: 105948, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36526034

RESUMO

Vibrio mimicus is a zoonotic pathogen that is widely distributed in aquatic habitats/environments (marine coastal water, estuaries, etc). The development of biocontrol agents for V. mimicus is imperative for the prevention and control of aquatic animal diseases and human food-borne infections. In this study, a broad-spectrum bacteriophage Vmp-1 was isolated from dealt aquatic product in a local market by double-layer agar plate method using V. mimicus CICC21613 as the host bacteria. Results indicated that Vmp-1, which belongs to the family Podoviridae, showed good pH tolerance (pH 3.0-12.0) and thermal stability (30-50 °C). The optimal multiplicity of infection (MOI) of Vmp-1 was 0.001 for a 20-min incubation and 100-min lysis period. Vmp-1 effectively controlled V. mimicus CICC21613 in LBS model (MOI = 0.0001, 0.001, 0.01, 0.1, 1) within 8 h. The full length of the Vmp-1 genome was 43,312 bp, with average GC content of 49.5%, and a total of 44 protein-coding regions. This study provides a novel phage strain that has the highest homology with vB_VpP_HA5 (GenBank: OK585159.1, 95.96%) for the development of biocontrol agents for V. mimicus.


Assuntos
Bacteriófagos , Vibrio mimicus , Vibrio , Animais , Humanos , Bacteriófagos/genética , Genômica , Vibrio/genética , Vibrio mimicus/genética , Proteínas de Membrana/metabolismo
5.
Polymers (Basel) ; 15(23)2023 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-38231920

RESUMO

The aim of this investigation was to scrutinize the effects of a thermal treatment on the electrostatic complex formed between gum arabic (GA) and ε-polylysine (ε-PL), with the goal of improving the antibacterial properties and reducing the hygroscopicity of ε-PL. The heated complex with a ratio of 1:4 exhibited an encapsulation efficiency of 93.3%. Additionally, it had an average particle size of 350.3 nm, a polydispersity index of 0.255, and a zeta potential of 18.9 mV. The formation of the electrostatic complex between GA and ε-PL was confirmed through multispectral analysis, which demonstrated the participation of hydrogen bonding and hydrophobic and electrostatic interactions, as well as the enhanced effect of heat treatment on these forces within the complex. The complex displayed a core-shell structure, with a regular distribution and a shape that was approximately spherical, as observed in the transmission electron microscopy images. Additionally, the heated GA-ε-PL electrostatic composite exhibited favorable antibacterial effects on Salmonella enterica and Listeria monocytogenes, with reduced minimum inhibitory concentrations (15.6 µg/mL and 62.5 µg/mL, respectively) and minimum bactericidal concentrations (31.3 µg/mL and 156.3 µg/mL, respectively) compared to free ε-PL or the unheated electrostatic composite. Moreover, the moisture absorption of ε-PL reduced from 92.6% to 15.0% in just 48 h after being incorporated with GA and subsequently subjected to heat. This research showed a way to improve the antibacterial efficiency and antihygroscopicity of ε-PL, reducing its application limitations as an antimicrobial substance to some extent.

6.
Food Res Int ; 160: 111623, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-36076436

RESUMO

This article describes the development of a novel liposome nanocarrier system. Carvacrol (Car) is first embedded in ß-cyclodextrin (ß-CD) by the freeze-drying method to form the ß-cyclodextrin-carvacrol inclusion compound (ß-CD-Car), and then ß-CD-Car liposomes (ß-CD-Car-LPs) and ß-CD-Car liposomes coated with S-layer proteins (SLPs) from Lactobacillus buchneri 20023 (SLP/ß-CD-Car-LPs) were prepared. The liposomes were characterized, and their stabilities, in vitro release characteristics, and antibacterial activities were investigated. Results showed that the fabricated liposome SLP/ß-CD-Car-LPs was nanosized, oval and homogenous, with the particle size of 229.1 ± 6.81 nm, the polydispersity index of 0.139, and the zeta potential of 27.9 mV. Measurements based on Triton X-100 resistance indicated that the SLP-coated liposomes were more stable than naked liposomes. The in vitro release study results showed that the rate of release from SLP-coated liposomes was much lower than that from uncoated liposomes. The minimum inhibitory activity (MIC) of SLP/ß-CD-Car-LPs (0.05 mg/mL) was 6.4 times higher than that of the free carvacrol (0.32 mg/mL) and was twice that of ß-CD-Car-LPs (0.1 mg/mL). In general, the stability, antibacterial activity, and sustained release effect of ß-CD-Car-LPs modified with SLPs were improved. Findings suggested that SLP-coated liposomes could be developed as a favorable delivery system for potential applications in the food industry.


Assuntos
Lipossomos , beta-Ciclodextrinas , Antibacterianos/farmacologia , Cimenos , Lactobacillus , Lipopolissacarídeos , Glicoproteínas de Membrana
7.
Front Microbiol ; 13: 830692, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35464931

RESUMO

Vibrio species are important pathogens of marine animals and aquaculture populations and some of them can cause serious infections in humans through consumption of contaminated seafood and aquaculture products. Lytic bacteriophages can potentially alleviate Vibrio contamination in the aquaculture organisms and in the processing of aquatic products and have gained significant scientific attention in recent years. In the present study, bacteriophages were isolated from sewage of local aquatic products markets and grown using Vibrio mimicus CICC 21613 as host cells. The lytic vibriophage OY1 belonging to the newly proposed family Autographiviridae and the genus Maculvirus was identified by observation under electron microscope and comparative genomic analysis. The phage OY1 showed lytic activity against 24 among 32 tested strains belonging to eight Vibrio species. The complete phage OY1 genome consists of a single circular double-stranded DNA of 43,479 bp with a total GC content of 49.27% and was predicted to encode 40 open reading frames (ORFs). To evaluate its potential against vibrios, the one-step growth curve, thermal and pH stability, host range, and lytic activity of the OY1 phage against Vibrio species were evaluated. The results showed that phage OY1 had a range of thermal and pH tolerance, and exhibited a significant inhibitory effect on the growth of tested Vibrio species. Bacterial growth in the fish muscle extract juice (FMEJ) inoculated with Vibrio mimicus CICC 21613, Vibrio parahaemolyticus CICC 21617, Vibrio alginolyticus VJ14, and the mixed bacterial culture was reduced by 2.65 log CFU/ml, 2.42 log CFU/ml, 1.93 log CFU/ml, and 2.01 log CFU/ml, respectively, by incubation with phage OY1 at 25°C for 36 h. Phage OY1 also showed a strong ability to prevent biofilm formation and destroy formed Vibrio species biofilms. These results indicate that phage OY1 is a potential biocontrol agent against Vibrio species in the aquaculture industry and in food safety control.

8.
J Hazard Mater ; 426: 128029, 2022 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-34942455

RESUMO

Various multi-drug-resistant microorganisms have appeared while a single antibacterial agent is increasingly no longer adequate for dealing with these resistant microorganisms. Herein, commercially purchased 50 nm-average-diameter silver nanoparticles (AgNPs) and Lactobacillus buchneri-isolated surface-layer proteins (SLPs) as a capping agent were used to fabricate a hybrid antibacterial agent (SLP-AgNPs) with enhanced antibacterial activity, and the possible synergistic antibacterial mechanism was explored. Characterization results revealed that SLP-AgNPs were uniformly surrounded by protein corona provided from SLP, and the formulations were mainly mediated by the electrostatic interactions and hydrogen bonding, which was evidenced by the results of Fourier transform infrared spectroscopy. According to the antibacterial tests, the minimum inhibitory concentration of SLP-AgNPs against Salmonella enterica (0.010 mg/mL) and Staphylococcus aureus (0.005 mg/mL) was 5-10 times lower than that of bare AgNPs, and while SLP-AgNPs showed a higher antibiofilm activity. Furthermore, bacterial cells exposed to SLP-AgNPs exhibited higher cell membrane permeability and stronger inhibition of respiratory-chain dehydrogenase activity, resulting in more severe cell death compared with bare AgNPs. The synergistic effect of SLP on AgNPs was probably carried out by enhanced function of adhesion to bacteria and antibacterial ability of SLP and SLP's supramolecular lattice structure on the sustained release of silver ion.


Assuntos
Nanopartículas Metálicas , Prata , Antibacterianos/farmacologia , Lactobacillus , Glicoproteínas de Membrana , Testes de Sensibilidade Microbiana , Espectroscopia de Infravermelho com Transformada de Fourier
9.
Microb Pathog ; 152: 104767, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33524565

RESUMO

Enterobacter hormaechei is a foodborne pathogen responsible for neonatal sepsis in humans and respiratory disease in animals. In this work, a new virulent phage (P.A-5) infecting E. hormaechei was isolated from domestic sewage samples and characterized. Transmission electron microscopy revealed that P.A-5 belonged to the family Myoviridae having a head size of 77.53 nm and a tail length of 72.24 nm. The burst size was 262 PFU/cell after a latent period of 20 min. Phage P.A-5 was able to survive in a pH range of 4-9 and resist temperatures up to 55 °C for 60 min. The genome sequence of P.A-5 had homology most similar to that of Shigellae phage MK-13 (GenBank: MK509462.1). Pork artificially contaminated with E. hormaechei was used as a model to evaluate the potential of P.A-5. The results clearly showed that P.A-5 treatment can completely inhibit E. hormaechei growth in pork within 8 h, indicating the potential use of P.A-5 as a biocontrol agent for E. hormaechei.


Assuntos
Bacteriófagos , Siphoviridae , Animais , Bacteriófagos/genética , Enterobacter , Genoma Viral , Genômica , Humanos , Recém-Nascido , Myoviridae/genética
10.
Food Funct ; 11(6): 5133-5141, 2020 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-32432306

RESUMO

The applications of carvacrol are limited due to its poor stability and water solubility, and high volatility; however, ovalbumin can be used to encapsulate hydrophobic molecules, improve their aqueous solubility, and reduce their volatility. In this study, we fabricated ovalbumin-carvacrol nanoparticles (OCGns) under different pH (2, 5, 7, and 9) conditions using a gel embedding method and investigated their physicochemical and antibacterial properties. Rheological experiments revealed that the G' of ovalbumin gels (OGs) prepared under different pH conditions were OG-2 > OG-7 > OG-9 > OG-5. Carvacrol addition reduced the tight structure of ovalbumin and carvacrol under pH 5 and 7 conditions, with hardness first decreasing and then increasing, but increasing under pH 2 and 9 conditions. Fluorescence and infrared spectroscopy indicated complex formation, with carvacrol increasing the average diameter of nanoparticles prepared at pH 2, 5, 7, and 9. Encapsulation reached 89.34 and 91.86% at pH 2 and 9, respectively; however, inhibition experiments revealed that the minimum inhibitory concentration of OCGn-2 against Gram-positive Bacillus cereus and Salmonella (0.08 and 0.16 mg mL-1, respectively) was lower than that of OCGn-9 (both 0.28 mg mL-1). Moreover, OCGn-2 possessed a better dense gel structure and a higher stability, encapsulation rate, and antibacterial activity, suggesting that pH affects gel microstructure and thus the encapsulation efficiency and bacteriostatic properties of the prepared nanoparticles. These results contribute to our knowledge of the design and fabrication of polymeric nanoparticle delivery systems for bioactive compounds with beneficial properties.


Assuntos
Antibacterianos/farmacologia , Bacillus cereus/efeitos dos fármacos , Cimenos/farmacologia , Nanogéis , Ovalbumina , Salmonella/efeitos dos fármacos , Cimenos/química , Concentração de Íons de Hidrogênio , Testes de Sensibilidade Microbiana , Nanocápsulas , Tamanho da Partícula , Reologia , Solubilidade
11.
Food Microbiol ; 86: 103330, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31703878

RESUMO

Pickle is a type of mildly lactic acid fermented vegetable and is a traditional dish favored in China, Japan, and Korea. Corruption of spoilage bacteria and accumulation of nitrite during vegetable fermentation are common problems that affect the pickle industry and consumer health. In this work, cucumber juice was used as a vegetable model to study the dominant mesophilic aerobic bacteria (MAB) producing nitrite during pickle fermentation. Virulent phages infecting the dominant MABs combined with Lactobacillus plantarum M6 were used to control these bacteria. Enterobacter cloacae and Pseudomonas fluorescens are the dominant MABs in the fermentation of cucumber juice containing 4% or 8% NaCl, with isolation percentages reaching 30.6% and 23.1%, respectively. Virulent phages PspYZU5415 and EcpYZU01 were isolated using P. fluorescens J5415 and E. cloacae J01 as the host bacteria, respectively. These two phages show a broad host range and strong lytic activity, and their genomes contain no toxins and antibiotic resistance genes. PspYZU5415 and EcpYZU01 were combined into a cocktail (designated as Phage MIX) that effectively inhibits the growth of E. cloacae and P. fluorescens in cucumber juice with different salt concentrations. PhageMIX combined with L. plantarum M6 decreased the counts of P. mendocina and E. cloacae to undetectable levels at 48 h during the fermentation of cucumber juice artificially contaminated with P. mendocina and E. cloacae. In addition, nitrite content increased to 11.3 mg/L at 20 h and then degraded completely at 36 h. By contrast, P. mendocina and E. cloacae remained in the groups without PhageMIX during fermentation (0-48 h). Nitrite content rapidly increased to 65.7 mg/L at 12 h and then decreased to 21.6 mg/L at 48 h in the control group. This study suggests that PhageMIX combined with lactic acid bacterial strains can be used as an ecological starter for controlling the dominant MABs P. mendocina and E. cloacae and for reducing nitrate production during the early stage of pickle fermentation.


Assuntos
Bacteriófagos/fisiologia , Bacteriófagos/patogenicidade , Cucumis sativus/microbiologia , Enterobacter cloacae/virologia , Microbiologia de Alimentos/métodos , Pseudomonas fluorescens/virologia , Verduras/microbiologia , Aerobiose , Bacteriófagos/genética , Bacteriófagos/isolamento & purificação , Cucumis sativus/metabolismo , Enterobacter cloacae/metabolismo , Fermentação , Alimentos Fermentados/microbiologia , Especificidade de Hospedeiro , Lactobacillus plantarum/metabolismo , Nitritos/metabolismo , Pseudomonas fluorescens/metabolismo
12.
Food Chem ; 308: 125590, 2020 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-31644970

RESUMO

A novel fluorescent nanoprobe was for the first time developed for the efficient detection of ferrocyanide ions ([Fe(CN)6]4-) based on nitrogen (N), sulfur (S) and chlorine (Cl) co-doped carbon nanoparticles (N,S,Cl-CNPs). The N,S,Cl-CNPs were fabricated through a simple and ultrafast acid-base neutralization method. The sensing mechanism was based on the quenching effect of [Fe(CN)6]4- on the fluorescence emission of N,S,Cl-CNPs via dynamic interaction. The N,S,Cl-CNPs were found to show high selectivity and sensitivity towards [Fe(CN)6]4- detection with two good linear relationships were achieved in the concentration ranges of 0.01-1.0 µg/mL and 1.0-50.0 µg/mL, respectively, and the detection limits are as low as 3.3 and 21.8 ng/mL, respectively. The proposed fluorescence method was successfully applied for [Fe(CN)6]4- analyses in food samples with high accuracy. The results of this study indicate the great application prospects of N,S,Cl-CNPs for [Fe(CN)6]4- detection in complex food matrix.


Assuntos
Carbono/química , Ferrocianetos/análise , Nanopartículas , Cloro/química , Ferrocianetos/química , Fluorescência , Análise de Alimentos , Limite de Detecção , Nitrogênio/química , Espectrometria de Fluorescência/métodos , Enxofre/química
13.
Carbohydr Polym ; 207: 362-370, 2019 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-30600018

RESUMO

Effect of pulsed electric field (PEF) on the structural properties of representative starches with different crystalline type, wheat starch for type A, potato starch for type B, and pea starch for type C, were investigated with polarized light microscopy (PLM), X-ray diffractometry (XRD), attenuated total internal reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, solid-state nuclear magnetic resonance (SSNMR) spectroscopy, small-angle X-ray scattering (SAXS), and gel permeation chromatography (GPC) to understand whether PEF could be applied directly in starchy foods. The results showed that PEF could change the structure of all three types of starch, especially potato starch; the birefringence, represented by Maltese cross in polarized microscopic observation changed slightly; XRD and SSNMR spectra demonstrated PEF did not change the crystalline type of starch granules. However, relative crystallinity variations happened at some points of electric field intensity (EFI). Increasing with the EFI, a bigger variation of R1045/1022 happened in potato starch than in wheat starch and pea starch, as illustrated by ATR-FTIR. Significant influences of PEF on the scatter structure and fractal dimension of self-similar structures were observed for wheat starch and potato starch, but not for pea starch. The GPC suggested that molecular weight distribution changed for all the three starches. And in vitro tests showed that PEF changed significantly (P < 0.05) the digestibility of starches, especially wheat starch and potato starch.

14.
Food Chem ; 280: 195-202, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-30642486

RESUMO

In this work, a novel simple, fast, selective and inexpensive fluorescence method for the determination of curcumin based on the fluorescence quenching of nitrogen and chlorine dual-doped carbon nanodots (N,Cl-CDs) was for the first time presented. The N,Cl-CDs were fastly and greenly produced by simply mixing glucose, 1,2-ethylenediamine (EDA) and hydrochloric acid (HCl). The fluorescence of N,Cl-CDs was significantly quenched by curcumin via a synergistic effect of dynamic quenching and inner filter effect (IFE). The N,Cl-CDs shows high selectivity and sensitivity towards curcumin sensing, achieving a linear range of 0.1-35 µM and a limit of detection (LOD) as low as 38 nM. The proposed fluorescence method was successfully utilized for curcumin detection in food samples with recoveries in a range of 96.8-106.1%. The findings of this study suggest the feasibility and usefulness of N,Cl-CDs as an effective fluorescence probe for the determination of curcumin in complex food matrix.


Assuntos
Carbono/química , Cloro/química , Curcumina/análise , Nitrogênio/química , Pontos Quânticos/química , Espectrometria de Fluorescência , Etilenodiaminas/química , Corantes Fluorescentes/química , Análise de Alimentos/métodos , Glucose/química , Ácido Clorídrico/química , Limite de Detecção
15.
Int J Food Microbiol ; 292: 107-117, 2019 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-30594742

RESUMO

The growth of Shewanella spp., mainly S. baltica and S. putrefaciens, is responsible for the spoilage of chilled fresh fish. Phages are an alternative tool to control bacterial growth. In this study, virulent phages infecting 4 S. baltica and 6 S. putrefaciens strains were isolated and characterized. Transmission electron microscopy revealed that 6 out of 10 phages (3 phages infecting S. baltica and 3 phages infecting S. putrefaciens) belonged to Myoviridae, while the other 4 phages (1 phage infecting S. baltica and 3 phages infecting S. putrefaciens) belonged to Siphoviridae. Phage SppYZU01 and SppYZU05 showed the broadest host range, being lytic towards all the 4 S. baltica strains and 5 out of the 6 S. putrefaciens strains, respectively. The genome sequence of SppYZU01 had no similarity with known genome sequences, while that of SppYZU05 was 88.5% similar to the genome of a virulent S. putrefaciens-infecting phage (Spp001). According to the host range and lytic activity, 3 phages, including SppYZU01, SppYZU05, and SppYZU06, were combined into a cocktail (designated as SPMIX3-156). SPMIX3-156 showed potential as an antimicrobial agent to control S. baltica and S. putrefaciens strain growth in catfish matrices. Bacterial growth in the catfish muscle juice inoculated with 104 colony-forming units (CFU)/mL of Shewanella strains was partially inhibited by 105 plaque-forming units (PFU)/mL of SPMIX3-156 at both 25 °C and 4 °C. The catfish fillets inoculated with Shewanella strains were used as a model to evaluate the biopreservative effects of SPMIX3-156. Total viable counts of fillet samples treated with 107 PFU/mL of SPMIX3-156 were reduced by 3.21 and 2.75 log units after 1 day at 25 °C and 10 day at 4 °C, respectively, compared to those of untreated samples. Fillet quality indices, including pH, total volatile basic nitrogen, and sensory value of the SPMIX3-156-treated samples, considerably improved compared to those of the control samples at both 4 °C and 25 °C. Our results suggest that SPMIX3-156 is a promising biological agent against S. baltica and S. putrefaciens, and may have a potential use in chilled fish fillet biopreservation.


Assuntos
Bacteriófagos/genética , Genes Virais , Ictaluridae/microbiologia , Shewanella putrefaciens/virologia , Shewanella/virologia , Animais , Bacteriófagos/isolamento & purificação , Agentes de Controle Biológico , Contagem de Colônia Microbiana , DNA Viral/isolamento & purificação , Contaminação de Alimentos , Microbiologia de Alimentos , Conservação de Alimentos , Myoviridae/isolamento & purificação , Alimentos Marinhos/microbiologia , Análise de Sequência de DNA , Siphoviridae/isolamento & purificação
16.
J Microbiol Biotechnol ; 27(11): 1971-1982, 2017 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-28910860

RESUMO

Piliated Lactobacillus rhamnosus (pLR) strains possess higher adherent capacity than non-piliated strains. The objective of this study was to isolate and characterize probiotic pLR strains in human fecal samples. To this end, mouse polyclonal antiserum (anti-SpaA) against the recombinant pilus protein (SpaA) of L. rhamnosus strain GG (LGG) was prepared and tested for its reactivity and specificity. With the anti-SpaA, a method combining the de Man, Rogosa, and Sharpe (MRS) agar plating separation and colony immunoblotting (CIB) was developed to isolate pLR from 124 human fecal samples. The genetic and phenotypic characteristics of the resultant pLR isolates were compared by randomly amplified polymorphic DNA (RAPD) fingerprinting, and examination of adhesion to Caco-2 cells, hydrophobicity, autoaggregation, and in vitro gastrointestinal tolerance. Anti-SpaA specifically reacted with three pLR strains of 25 test strains, as assessed by western blotting, immunofluorescence flow cytometry, and immunoelectron microscopy (IEM) assays. The optimized MRS agar separation plus anti-SpaA-based CIB procedure could quantitatively detect 2.5 × 103 CFU/ml of pLR colonies spiked in 106 CFU/ml of background bacteria. Eight pLR strains were identified in 124 human fecal samples, and were confirmed by 16S RNA gene sequencing and IEM identification. RAPD fingerprinting of the pLR strains revealed seven different patterns, of which only two isolates from infants showed the same RAPD profiles with LGG. Strain PLR06 was obtained with high adhesion and autoaggregation activities, hydrophobicity, and gastrointestinal tolerance. Anti-SpaA-based CIB is a rapid and inexpensive method for the preliminary screening of novel adherent L. rhamnosus strains for commercial purposes.


Assuntos
Fezes/microbiologia , Proteínas de Fímbrias/imunologia , Microbioma Gastrointestinal/fisiologia , Soros Imunes/imunologia , Immunoblotting/métodos , Lacticaseibacillus rhamnosus/imunologia , Lacticaseibacillus rhamnosus/isolamento & purificação , Probióticos , Ácidos , Animais , Aderência Bacteriana/efeitos dos fármacos , Aderência Bacteriana/imunologia , Ácidos e Sais Biliares/farmacologia , Western Blotting , Células CACO-2 , Análise por Conglomerados , Reações Cruzadas/imunologia , Microbioma Gastrointestinal/efeitos dos fármacos , Microbioma Gastrointestinal/imunologia , Humanos , Interações Hidrofóbicas e Hidrofílicas , Lacticaseibacillus rhamnosus/efeitos dos fármacos , Lacticaseibacillus rhamnosus/genética , Proteínas de Membrana/química , Proteínas de Membrana/imunologia , Camundongos , Subunidades Proteicas , RNA Ribossômico 16S/genética , Técnica de Amplificação ao Acaso de DNA Polimórfico , Proteínas Recombinantes/imunologia
17.
Food Chem Toxicol ; 50(5): 1238-44, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22300772

RESUMO

Evaluation of the chemical composition and antihyperglycemic and antioxidant activity of five wild edible mushrooms (Clitocybe maxima, Catathelasma ventricosum, Stropharia rugoso-annulata, Craterellus cornucopioides and Laccaria amethystea) from Southwest China. The chemical composition assay includes proximate analysis (moisture, ash, crude protein, crude fat, total carbohydrates and total energy), bioactive compounds analysis (total phenolic, flavonoid, ascorbic acid, ergosterol, tocopherol), fatty acid analysis, amino acid analysis, phenolic compounds analysis and mineral analysis of these mushrooms. Furthermore, assays of α-glucosidase inhibitory and α-amylase inhibitory activity were used for evaluating antihyperglycemic activity of the mushrooms, and assays of reducing power, chelating effect on ferrous ions, scavenging effect on hydroxyl free radicals and 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity were used for evaluating antioxidant activity of the mushrooms. Based on the results, ethanolic and aqueous extract of these mushroom all showed antihyperglycemic and antioxidant potential. In particular, the aqueous extract of C. ventricosum revealed the highest α-glucosidase inhibitory activity (EC50 value 2.74 µg/mL), DPPH radical scavenging activity (EC50 value 2.86 mg/mL) and reducing power (EC50 value 0.96 mg/mL), while the aqueous extract of L. amethystea showed the highest α-amylase inhibitory activity (EC50 value 4.37 µg/mL) and metal chelating activity (EC50 value 2.13 mg/mL).


Assuntos
Agaricales/química , Antioxidantes/uso terapêutico , Hipoglicemiantes/uso terapêutico , Agaricales/classificação , Antioxidantes/farmacologia , China , Cromatografia Líquida de Alta Pressão , Avaliação Pré-Clínica de Medicamentos , Inibidores de Glicosídeo Hidrolases , Hipoglicemiantes/farmacologia , Especificidade da Espécie , alfa-Amilases/antagonistas & inibidores
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