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1.
Front Microbiol ; 14: 1201201, 2023.
Article in English | MEDLINE | ID: mdl-37538844

ABSTRACT

Listeria monocytogenes (L. monocytogenes) is a foodborne pathogen that poses significant risks to public health and food safety. The present study aimed to identify the presence of Listeria spp. in various samples, including pasteurized milk, chicken fillets, and stool samples from pregnant women in Sharkia Governorate, Egypt. Additionally, the study identified the serotypes, virulence-associated genes, antimicrobial resistance patterns, and biofilm formation in L. monocytogenes isolates. Moreover, the antibacterial and anti-biofilm activity of Lactobacillus plantarum ATCC 14917 (L. plantarum) against L. monocytogenes isolates was investigated. A cross-sectional study was conducted from August 2021 to January 2022 to collect 300 samples of pasteurized milk, chicken fillets, and stool from pregnant women admitted to outpatient clinics of hospitals. The results showed that 32.7% of the samples were positive for Listeria spp., including L. innocua (48.9%), L. monocytogenes (26.5%), L. ivanovii (14.3%), L. grayi (5.1%), and L. welshimeri (5.1%). Among all L. monocytogenes isolates, hlyA, actA, inlC, and inlJ virulence-associated genes were detected. However, the virulence genes plcB, iap, and inlA were found in 10 (38.5%), 8 (30.8%), and 25 (96.2%) isolates, respectively. The L. monocytogenes isolates classified into four serotypes (1/2a, 1/2b, 1/2c, and 4b), with 1/2a and 4b each identified in 30.8% of the isolates, while 1/2b and 1/2c were identified in 19.2% of the isolates. All L. monocytogenes isolates showed 100% resistance to streptomycin, kanamycin, and nalidix acid, and 92.3% of isolates showed gentamicin resistance. However, all isolates were susceptible to ampicillin and ampicillin/sulbactam. Multidrug resistance (MDR) was observed in 20 (76.9%) L. monocytogenes isolates. The biofilm formation ability of 26 L. monocytogenes isolates was evaluated at different incubation temperatures. At 4°C, 25°C, and 37°C, 53.8, 69.2, and 80.8% of the isolates, respectively, were biofilm producers. Furthermore, 23.1% were strong biofilm producers at both 4°C and 25°C, while 34.6% were strong biofilm formers at 37°C. Treating L. monocytogenes isolates with L. plantarum cell-free supernatant (CFS) reduced the number of biofilm-producing isolates to 15.4, 42.3, and 53.8% at 4°C, 25°C, and 37°C, respectively. L. plantarum's CFS antibacterial activity was tested against six virulent, MDR, and biofilm-forming L. monocytogenes isolates. At a concentration of 5 µg/mL of L. plantarum CFS, none of the L. monocytogenes isolates exhibited an inhibition zone. However, an inhibition zone was observed against L. monocytogenes strains isolated from pasteurized milk and pregnant women's stools when using a concentration of 10 µg/mL. Transmission electron microscopy (TEM) revealed that L. plantarum CFS induced morphological and intracellular structural changes in L. monocytogenes. In conclusion, this study identified virulent MDR L. monocytogenes isolates with strong biofilm-forming abilities in food products in Egypt, posing significant risks to food safety. Monitoring the prevalence and antimicrobial resistance profile of L. monocytogenes in dairy and meat products is crucial to enhance their safety. Although L. plantarum CFS showed potential antibacterial and anti-biofilm effects against L. monocytogenes isolates, further research is needed to explore its full probiotic potential.

2.
Nanomaterials (Basel) ; 11(11)2021 Nov 09.
Article in English | MEDLINE | ID: mdl-34835774

ABSTRACT

The use of nanomaterials alone or in composites with proteins is a promising alternative to inhibit pathogenic bacteria. In this regard, this study used seed proteins from both fenugreek (Trigonella foenum-graecum L.) (FNP) and mung bean (Viga radiate) (MNP), with silver nanoparticles (Ag-NPs) and nanocomposites of either Ag-NPs plus FNP (Ag-FNP) or Ag-NPs plus MNP (Ag-MNP) as inhibitory agents against pathogenic bacteria. FNP and MNP were isolated from fenugreek seeds and mung bean seeds, respectively, and fractionated using Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). Both FNP and MNP were immobilized with Ag-NPs to synthesize the nanocomposites Ag-FNP and Ag-MNP, respectively. The physicochemical characteristics of Ag-NPs and their composites with proteins were studied by X-ray Diffraction (XRD), dynamic light scattering (DLS), the zeta potential, Scanning and Transmission Electron Microscopy (SEM and TEM, respectively), Atomic Force Microscopy (AFM), and the Brunauer-Emmett-Teller isotherm (BET), elucidating their structural parameters, size distribution, size charges, size surface morphology, particle shape, dimensional forms of particles, and specific surface area, respectively. The sole proteins, Ag-NPs, and their nanocomposites inhibited pathogenic Gram-positive and Gram-negative bacteria. The inhibitory activities of both nanocomposites (Ag-FNP and Ag-MNP) were more than those obtained by either Ag-NPs or proteins (FNP, MNP). Minimum inhibitory concentrations (MICs) of Ag-FNP were very low (20 and 10 µg mL-1) against Salmonellatyphimurium and Pseudomonasaerugenosa, respectively, but higher (162 µg mL-1) against E. coli and Listeriamonocytogenes. MICs of Ag-MNP were also very low (20 µg mL-1) against Staphylococcusaureus but higher (325 µg mL-1) against Listeriamonocytogenes. TEM images of Staphylococcusaureus and Salmonellatyphimurium, treated with Ag-FNP and Ag-MNP, at their MIC values, showed asymmetric, wrinkled exterior surfaces, cell deformations, cell depressions, and diminished cell numbers.

3.
Antibiotics (Basel) ; 10(7)2021 Jul 17.
Article in English | MEDLINE | ID: mdl-34356791

ABSTRACT

Cowpea seed protein hydrolysates (CPH) were output from cowpea seeds applying alcalase® from Bacillus licheniformis. CPH with an elevated level of hydrolysis was fractionated by size exclusion chromatography (SEC). Both CPH and SEC-portions showed to contain antimicrobial peptides (AMPs) as they inhibited both Gram-positive bacteria, such as Listeria monocytogenes LMG10470 (L. monocytogenes), Listeria innocua. LMG11387 (L. innocua), Staphylococcus aureus ATCC25923 (S.aureus), and Streptococcus pyogenes ATCC19615 (St.pyogenes), and Gram-negative bacteria, such as Klebsiella pnemoniae ATCC43816 (K. pnemoniae), Pseudomonas aeroginosa ATCC26853 (P. aeroginosa), Escherichia coli ATCC25468) (E.coli) and Salmonella typhimurium ATCC14028 (S. typhimurium).The data exhibited that both CPH and size exclusion chromatography-fraction 1 (SEC-F1) showed high antibacterial efficiency versus almost all the assessed bacteria. The MIC of the AMPs within SEC-F1 and CPHs were (25 µg/mL) against P. aeruginosa, E.coli and St. pyogenes. However, higher MICsof approximately 100-150 µg/mL showed for both CPHs and SEC-F1 against both S. aureus and L. innocua; it was 50 µg/mL of CPH against S.aureus. The Electro-spray-ionization-mass-spectrometry (ESI-MS) of fraction (1) revealed 10 dipeptides with a molecular masses arranged from 184 Da to 364 Da and one Penta peptide with a molecular mass of approximately 659 Da inthe case of positive ions. While the negative ions showed 4 dipeptides with the molecular masses that arranged from 330 Da to 373 Da. Transmission electron microscope (TEM) demonstrated that the SEC-F1 induced changes in the bacterial cells affected. Thus, the results suggested that the hydrolysis of cowpea seed proteins by Alcalase is an uncomplicated appliance to intensify its antibacterial efficiency.

4.
Molecules ; 26(16)2021 Aug 19.
Article in English | MEDLINE | ID: mdl-34443614

ABSTRACT

Kombucha is a traditional beverage of sweetened black tea fermented with a symbiotic association of acetic acid bacteria and yeasts. In this study, kombucha fermented beverage (KFB) appeared to include nine chemical groups (alcohols, acids, lactones, condensed heterocyclic compounds, antibiotics, esters, aldehydes, fatty acids, and alkaloids) of many bioactive metabolites, as elucidated by gas chromatography-mass spectrometry (GC-MS) and IR spectra. The fermented metabolic components of KFB seem collectively to act in a synergistic action giving rise to the antimicrobial activity. Four types of kombucha preparations (fermented, neutralized, heat-treated and unfermented) were demonstrated with respect to their antimicrobial activity against some pathogenic bacterial and fungal strains using agar well diffusion assay. KFB exerted the strongest antimicrobial activities when compared with neutralized and heat-treated kombucha beverages (NKB and HKB). Staphylococcus aureus ATCC6538 (S. aureus) and Escherichia coli ATCC11229 (E. coli) were the organisms most susceptible to the antimicrobial activity of kombucha beverage preparations. Finally, the KFB preparation showed remarkable inhibitory activity against S. aureus and E. coli bacteria in a brain heart infusion broth and in some Egyptian fruit juices (apple, guava, strawberry, and tomato). These data reveal that kombucha is not only a prophylactic agent, but also appears to be promising as a safe alternative biopreservative, offering protection against pathogenic bacteria and fungi.


Subject(s)
Anti-Infective Agents/analysis , Anti-Infective Agents/pharmacology , Fermented Foods/analysis , Fermented Foods/microbiology , Hydrogen-Ion Concentration
5.
Molecules ; 26(9)2021 Apr 30.
Article in English | MEDLINE | ID: mdl-33946451

ABSTRACT

Kefir beverage (KB) is a fermented milk initiated by kefir grains rich with starter probiotics. The KB produced in this study seemed to contain many chemical compounds elucidated by gas chromatography-mass spectrometry (GC-MS) and IR spectra. These compounds could be classified into different chemical groups such as alcohols, phenols, esters, fatty esters, unsaturated fatty esters, steroids, polyalkenes, heterocyclic compounds and aromatic aldehydes. Both KB and neutralized kefir beverage (NKB) inhibited some pathogenic bacteria including Escherichia coli ATCC11229 (E. coli), Listeria monocytogenes ATCC 4957 (L. monocytogenes), Bacillus cereus ATCC 14579 (B. cereus), Salmonella typhimurium ATCC 14028 (Sal. typhimurium) as well as some tested fungal strains such as Aspergillus flavus ATCC 16872 (A. flavus) and Aspergillus niger ATCC 20611 (A. niger), but the inhibitory activity of KB was more powerful than that obtained by NKB. It also appeared to contain four lactic acid bacteria species, one acetic acid bacterium and two yeast species. Finally, the KB inhibited distinctively both S. aureus and Sal. typhimurium bacteria in a brain heart infusion broth and in some Egyptian fruit juices, including those made with apples, guava, strawberries and tomatoes.


Subject(s)
Anti-Infective Agents/chemistry , Anti-Infective Agents/pharmacology , Kefir/analysis , Fermentation , Fermented Foods , Food Analysis , Gas Chromatography-Mass Spectrometry , Hydrogen-Ion Concentration , Molecular Structure , Temperature
6.
Antibiotics (Basel) ; 9(12)2020 Dec 13.
Article in English | MEDLINE | ID: mdl-33322196

ABSTRACT

Native egg albumin (NEA) was isolated from hen eggs and hydrolyzed by pepsin to produce hydrolyzed egg albumin (HEA). HEA was chemically characterized and screened for its antibacterial activity against 10 pathogenic bacteria (6 Gram (+) and 4 Gram (-)). The SDS-PAGE pattern of NEA showed molecular weights of hen egg albumin subunits ranging from 30 to 180 kDa. The highest intensive bands appeared at a molecular mass of about 50 and 97 kDa. Ultra-performance liquid chromatography (UPLC) of the peptic HEA revealed 44 peptides, 17 of them were dipeptides, and the other 27 fractions corresponded to bigger peptides (3-9 amino acids). The dipeptides and big peptides represented 26% and 74% of the total hydrolysate, respectively. The MIC of HEA was about 100 µg/L for Listeria monocytogenes, Bacillus cereus, Staphylococcus aureus, Salmonella typhimurium, Streptococcus pyogenes, and Klebsiella oxytoca and 150 µg/L for Pseudomonas aeruginosa, Bacillus subtilis, and Listeria ivanovii and 200 µg/L for Escherichia coli. L. monocytogenes was the most sensitive organism to HEA. Mixtures of HEA with antibiotics showed more significant antibacterial activity than individually using them. Transmission electron microscopy (TEM) revealed various signs of cellular deformation in the protein-treated bacteria. HEA may electrostatically and hydrophobically interact with the cell wall and cell membrane of the susceptible bacteria, engendering large pores and pore channels leading to cell wall and cell membrane disintegration. Higher cell permeability may, thus, occur, leading to cell emptiness, lysis, and finally death. Alternatively, no toxicity signs appeared when HEA was administrated to Wistar Albino rats as one single dose (2000, 5000 mg/kg body weight) or repeated daily dose (500 and 2500 mg/kg body weight/day) for 28 days to disclose the possible toxicity hazards. HEA did not produce any death.

7.
Molecules ; 25(20)2020 Oct 20.
Article in English | MEDLINE | ID: mdl-33092293

ABSTRACT

Penicillium chrysogenum has been reported as a potent taxol producer based on quantitative analysis by TLC and HPLC. The biosynthetic potency of taxol has been validated from PCR detection of rate-limiting genes of taxol synthesis such as taxadienesynthase and 10-de-acetylbaccatin III-O-acetyltransferase (DBAT), which catalyzes the immediate diterpenoid precursor of the taxol substance, as detected by PCR. Taxol production by P. chrysogenum was assessed by growing the fungus on different media. Potato dextrose broth (PDB) was shown to be the best medium for obtaining the higher amount of taxol (170 µg/L). A stepwise optimization of culture conditions necessary for production of higher amounts of taxol was investigated. The substance taxol was produced optimally after 18 d of incubation at 30 °C in PDB adjusted initially at pH 8.0 with shaking (120 rpm) (250 µg/L). The P. chrysogenum taxol was purified successfully by HPLC. Instrumental analyzes such as Fourier transform infrared spectroscopy (FTIR), ultraviolet (UV) spectroscopy, 1HNMR and 13C NMR approved the structural formula of taxol (C47H51NO14), as constructed by ChemDraw. The P. chrysogenum taxol showed promising anticancer activity.


Subject(s)
Cell Proliferation/drug effects , Paclitaxel/chemistry , Penicillium chrysogenum/chemistry , Chromatography, High Pressure Liquid , Humans , Isomerases/biosynthesis , Isomerases/chemistry , Magnetic Resonance Spectroscopy , Molecular Structure , Paclitaxel/biosynthesis , Paclitaxel/isolation & purification , Paclitaxel/pharmacology , Penicillium chrysogenum/enzymology , Spectroscopy, Fourier Transform Infrared
8.
Molecules ; 25(19)2020 Oct 07.
Article in English | MEDLINE | ID: mdl-33036497

ABSTRACT

Bacterial outbreaks caused by Staphylococcus aureus (S. aureus) are interesting due to the existence of multidrug resistant (MDR) isolates. Therefore, there is a need to develop novel ways to control such MDR S. aureus. In this study, some natural agents such as honey bee (HB), extracts of either Moringa oleifera seeds (MSE), or leaves (MLE) and essential oils of garlic, clove, and moringa were studied for their inhibitory activity against this S. aureus pathogen. About 100 food samples including beef luncheon (n = 25), potato chips (n = 50), and corn flakes (n = 25) were investigated for possible pollution with the S. aureus bacteria. The isolated bacteria suspected to belong S. aureus that grew well onto Baird-Parker agar (Oxoid) and shiny halo zones and positive coagulase reaction were selected and identified by API-Kits; all of them that were approved belong to S. aureus (18 strains). The sensitivity of the obtained 18 S. aureus bacterial strains to 12 antibiotics were evaluated; all of them were resistant to ofloxacin; however, other antibiotics tested showed variable results. Interestingly, the S. aureus No. B3 isolated from beef luncheon was resistant to10 antibiotics out of 12 ones tested. Multiple antibiotic resistance index (MAR) of this S. aureus strain was about 83.3%. Therefore, its identification was confirmed by sequencing of a 16S rRNA gene which approved a successful biochemical identification carried out by API Kits and such strain was designated S. aureus LC 554891. The genome of such strain appeared to contain mecA gene encoding methicillin resistance; it was found to contain hla, hlb, tsst-1, and finbA that encode α-blood hemolysis, ß-blood hemolysis, toxic shock syndrome gene, and fibrinogen-binding protein gene, respectively. In addition, the virulence factors viz. sea; seb; sec encoding enterotoxins were detected in the DNA extracted from S. aureus B3 strain. Aqueous extract of Moringa oleifera seeds (MSE) showed inhibitory activity against S. aureus LC 554891 better than that obtained by tetracycline, essential oils or HB. Minimum inhibitory concentration (MIC) of MSE was 20µg/mL. Instrumental analysis of MSE showed 14 bioactive chemical compounds. Combinations of both MSE and tetracycline showed distinctive inhibitory activity against S. aureus LC 554891 than that obtained by either tetracycline or MSE singly.


Subject(s)
Anti-Bacterial Agents/pharmacology , Moringa oleifera/chemistry , Staphylococcus aureus/drug effects , Anti-Bacterial Agents/chemistry , Microbial Sensitivity Tests , RNA, Ribosomal, 16S/genetics , RNA, Ribosomal, 16S/metabolism , Virulence Factors
9.
Foods ; 9(8)2020 Aug 13.
Article in English | MEDLINE | ID: mdl-32823600

ABSTRACT

Minced beef is a very perishable food product, due to its vulnerability to microbial contamination and its fast quality deterioration. In the current study, the biological efficiency of different concentrations (0, 50 and 100 µg g-1) of the antibacterial catfish glycoprotein (CFG) was estimated as a possible improver of the storability and safety of minced beef preserved at 4 °C for 15 days. CFG (50 and 100 µg g-1) could efficiently control the changes in meat pH during 15 days storage at 4 °C to be within the normal, acceptable levels (6.4 and 6.2, respectively), equalizing the level of the control for minced beef after 6 days of storage under similar conditions. Likewise, the level of metmyoglobin in minced beef stored at the same conditions was maintained at 53.67 and 46.67% by CFG supplementation at 50 and 100 µg g-1, respectively, at the 15th day of storage, which is comparable to the 6th day in case of the control samples. However, the antioxidant effect of CFG against lipid peroxidation was less effective. The antibacterial action of CFG was most pronouncedly powerful and efficient. Supplementation of minced beef with CFG at 50 and 100 µg g-1 significantly (p < 0.05) decreased the bacterial counts at all the time inspection points as compared to the control. After 15 days of storage, the total viable bacteria, psychrotrophic bacterial count and coliforms count were reduced to 3.12, 2.65 and 0.0 log CFU g-1, respectively, in response to CFG (50 µg g-1), and 2.41, 2.04 and 0.0 log CFU g-1, respectively, in response to CFG (100 µg g-1); this compared to 5.13, 4.78 and 2.5 in the control samples after only six days cold storage. Using CFG at 50, 100 and 200 µg g-1 in rat diets did not affect their liver or kidney functions, reflecting the non-toxicity of this substance. Substantiating the antioxidant and antimicrobial potential of CFG in minced beef storage may support its use as a naturally powerful and safe food preservative, as well as a shelf-life extender.

10.
Antibiotics (Basel) ; 9(4)2020 Apr 13.
Article in English | MEDLINE | ID: mdl-32294942

ABSTRACT

There is a need to continue research to find out other anti-dermatophytic agents to inhibit causal pathogenic skin diseases including many types of tinea. We undertook the production, purification, and identification of an anti-dermatophytic substance by Streptomyces atrovirens. Out of 103 streptomycete isolates tested, only 20 of them showed antidermatophytic activity with variable degrees against Trichophyton tonsurans CCASU 56400 (T. tonsurans), Microsporum canis CCASU 56402 (M. canis), and Trichophyton mentagrophytes CCASU 56404 (T. mentagrophytes). The most potent isolate, S10Q6, was identified based on the tests conducted that identified morphological and physiological characteristics and using 16S rRNA gene sequencing. The isolate was found to be closely correlated to previously described species Streptomyces atrovirens; it was designated Streptomyces atrovirens KM192347 (S. atrovirens). Maximum antifungal activity of the strain KM192347 was obtained in modified starch nitrate medium (MSNM) adjusted initially at pH 7.0 and incubated at 30 °C in shaken cultures (150 rpm) for seven days. The antifungal compound was purified by using two steps protocol including solvent extraction and column chromatography. The MIC of it was 20µg/mL against the dermatophyte cultures tested. According to the data obtained from instrumental analysis and surveying the novel antibiotics database, the antidermatophytic substance produced by the strain KM192347 was characterized as an oxaborole-6-benzene sulphonamide derivative and designated oxaborole-6-benzene sulphonamide (OXBS) with the chemical formula C13H12 BNO4S. The crude OXBS didn't show any toxicity on living cells. Finally, the results obtained herein described another anti-dermatophytic substance named an OXBS derivative. .

11.
Molecules ; 24(23)2019 Nov 24.
Article in English | MEDLINE | ID: mdl-31771271

ABSTRACT

Crude, phenolic-rich extracts (CPREs) were isolated from different sources, such as Hibiscus sabdariffa (H. sabdariffa), Brassica oleracea var. capitata f. rubra (B. oleracea) and Beta vulgaris (B. vulgaris) and characterized. These CPREs showed potential antibacterial and antifungal activities. H. sabdariffa CPRE (HCPRE) is the most potent, as it inhibited all tested bacteria and fungi. Total anthocyanins content (TAC), total phenolic content (TPC) and total flavonoid content (TFC) were estimated in all three CPREs. H. sabdariffa contained 4.2 mg/100 g TAC, 2000 mg/100 g of TPC and 430 mg/100 g of TFC in a dry weight sample. GC-MS analysis of HCPRE showed 10 different active compounds that have antimicrobial effects against pathogenic bacteria and fungi, especially alcoholic compounds, triazine derivatives and esters. Scanning and transmission electron microscopy images of Staphylococcus aureus DSM 1104 and Klebsiella pneumonia ATCC 43816 treated with HCPRE (50 µg/mL) exhibited signs of asymmetric, wrinkled exterior surfaces, cell deformations and loss of cell shapes; and adherence of lysed cell content led to cell clumping, malformations, blisters, cell depressions and diminished cell numbers. This indicates death of bacterial cells and loss of cell contents. Aspergillus ochraceus EMCC516 (A. ochraceus, when treated with 100 µg/mL of HCPRE showed irregular cell organelles and cell vacuolation.


Subject(s)
Anti-Bacterial Agents/pharmacology , Antifungal Agents/pharmacology , Beta vulgaris/chemistry , Brassica/chemistry , Hibiscus/chemistry , Phenols/pharmacology , Anthocyanins/chemistry , Anthocyanins/pharmacology , Anti-Bacterial Agents/chemistry , Antifungal Agents/chemistry , Aspergillus ochraceus/drug effects , Flavonoids/chemistry , Flavonoids/pharmacology , Gas Chromatography-Mass Spectrometry , Klebsiella pneumoniae/drug effects , Microscopy, Electron, Transmission , Molecular Structure , Phenols/chemistry , Plant Extracts/chemistry , Plant Extracts/pharmacology , Staphylococcus aureus/drug effects
12.
Molecules ; 24(16)2019 Aug 09.
Article in English | MEDLINE | ID: mdl-31405067

ABSTRACT

Streptococcus pyogenes (S. pyogenes) ZUH1 was isolated and characterized using morphological, cultural and biochemical methods. The results showed that the marker genes (namely spyCEP, ssa, sic, sdaB and speG) indicating group A streptococci (GAS) were detected in the S. pyogenes genome. The results showed that the S. pyogenes strain was inhibited by Crocus sativus methanol extract (CSME), bee honey (BH) and catfish glycoprotein (CFG). The inhibitory activity of these natural agents were compared with standard antibiotics such as Ceftazidime (30 µg/mL), Cefoperazone (75 µg/mL), Cefoxitin (30 µg/mL) and Imipenem (10 µg/mL). There was a synergistic effect between certain antibiotics and CSME. GC-MS and IR analysis of CSME showed different cyclic ketones, aldehydes, esters, alcohols and acids. The main compounds were tetradecanoic acid, safranal and isophorone. Transmission electron microscopy (TEM) images of S. pyogenes cells treated with CSME showed signs of an irregular wrinkled outer surface, fragmentation, adhesion and aggregation of damaged bacterial cells or cellular debris. The marker genes (spyCEP, ssa, sic, sdaB and speG) could be used as a rapid diagnostic tool for GAS. CSME, BH and CFG showed distinctive anti-streptococcal activity either alone or in combinations with antibiotics; their action on S. pyogenes cells was studied by TEM. There was a synergistic effect between antibiotics and Crocus sativus, bee honey, and glycoprotein against S. pyogenes ZUH1. The action of natural agents on the pathogenic cells was shown using TEM.


Subject(s)
Anti-Bacterial Agents , Crocus/chemistry , Honey , Plant Extracts , Streptococcal Infections/drug therapy , Streptococcus pyogenes/growth & development , Uveitis, Intermediate/drug therapy , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Drug Therapy, Combination , Humans , Male , Methanol/chemistry , Middle Aged , Plant Extracts/chemistry , Plant Extracts/pharmacology , Streptococcal Infections/metabolism , Streptococcal Infections/pathology , Uveitis, Intermediate/microbiology , Uveitis, Intermediate/pathology
13.
Int J Biol Macromol ; 138: 773-780, 2019 Oct 01.
Article in English | MEDLINE | ID: mdl-31351952

ABSTRACT

Catfish glycoprotein (CFG) was extracted from the cutaneous mucus of Egyptian catfish by ammonium sulphate precipitation and purified on gel filtration column (sephadex G-50). After purification, CFG produced one band on SDS-PAGE (22 kDa). Urea-PAGE and the pH-solubility of CFG indicated its positive charge (IEP 8). CFG contained 12 saccharides. FTIR spectrum shows 3 groups of bands at 1800-2900, 1100-1700 and 700-1100 cm-1. CFG exhibited antibacterial activity against 9 pathogenic bacteria with low MIC (50 µg/mL), where two Gram+ bacteria, i.e.; Streptococcus pyogenes (St. pyogenes) and Listeria ivanovii (L. ivanovii) were the most sensitive. The growth curves of the bacteria subjected to 1 MIC of CFG during 30 h showed general growth inhibition, particularly in case of Gram- bacteria such as E. coli. TEM images showed evidently reduced relative content of the intact cells and clear incurred cellular malformations. Combining CFG with specific antibiotic at equal ratios induced synergistic antibacterial actions, amounting to 40% of the mathematical sum of the combination. Substituting the antibiotic chloramphenicol with gradual increasing ratios of CFG of its starting concentration (30 µg/mL), produced proportionally bigger antibacterial actions against St. pyogenes growth and increasing synergistic effect up to 37% at 80% of CFG substitution.


Subject(s)
Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Catfishes , Glycoproteins/chemistry , Glycoproteins/pharmacology , Mucus/chemistry , Animals , Anti-Bacterial Agents/isolation & purification , Catfishes/metabolism , Dose-Response Relationship, Drug , Glycoproteins/isolation & purification , Microbial Sensitivity Tests , Spectrum Analysis , Structure-Activity Relationship
14.
Molecules ; 24(6)2019 Mar 19.
Article in English | MEDLINE | ID: mdl-30893826

ABSTRACT

The present work was carried out to determine the characteristics and antibacterial activity of 7S and 11S globulins isolated from cowpea seed (Vigna unguiculata (L.) Walp.). The molecular mass of 7S globulin was demonstrated by SDS-PAGE bands to be of about 132, 129 and 95 kDa corresponding the α/, α and ß subunits. The molecular mass of 11S globulin was demonstrated by SDS-PAGE bands to be existed between 28 and 52 kDa corresponding the basic and acidic subunits. The minimum inhibitory concentrations MICs of 7S and 11S globulins isolated from cowpea seed were determined against Gram positive bacteria viz: Listeria monocytogenes LMG 10470, Listeria ivanovii FLB 12, Staphylococcus aureus ATCC 25923 and Streptococcus pyogenes ATCC 19615, and Gram negative bacteria such as Klebsiella pneumonia ATCC 43816, Pseudomonas aeruginosa ATCC 26853, Escherichia coli ATCC 25922 and Salmonella ATCC 14028 using disc diffusion assay; they were showed to be in the range 10 to 200 µg/mL. Transmission electron microscope (TEM) examination of the protein-treated bacteria showed the antibacterial action of 11S globulin against S. typhimurium and P. aeruginosa was manifested by signs of cellular deformation, partial and complete lysis of cell components. Adding 11S globulin at both concentrations 50 and 100 µg/g to minced meat showed considerable decreases in bacterial counts of viable bacteria, psychrotrophs and coliforms compared to controls during 15 days storage at 4 °C, reflecting a promising perspective to use such globulin as a meat bio-preservative.


Subject(s)
Globulins/pharmacology , Seed Storage Proteins/pharmacology , Vigna/chemistry , Anti-Bacterial Agents/pharmacology , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Meat/microbiology , Microbial Sensitivity Tests , Microscopy, Electron, Transmission
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