Mapping pathogenic bacteria resistance against common antibiotics and their potential susceptibility to methylated white kidney bean protein.

As antibiotics cannot inhibit multidrug-resistant bacteria (MDR), continuous research is mandatory to find other antibacterials from natural resources. Native legume proteins and their modified forms exhibited broad spectra of high antimicrobial activities. Sixteen bacterial isolates were mapped for antibiotic resistance, showing resistance in the range of (58-92%) and (42-92%) in the case of the Gram-negative and Gram-positive bacteria, respectively. White native Phaseolus vulgaris protein (NPP) was isolated from the seeds and methylated (MPP). The MIC range of MPP against 7 MDR bacteria was 10-25 times lower than NPP and could (1 MIC) considerably inhibit their 24 h liquid growth. MPP showed higher antibacterial effectiveness than Gentamycin, the most effective antibiotic against Gram-positive bacteria and the second most effective against Gram-negative bacteria. However, MPP recorded MICs against the seven studied MDR bacteria in the 1-20 µg/mL range, the same for Gentamycin. The combination of Gentamycin and MPP produced synergistic effects against the seven bacteria studied, as confirmed by the Transmission Electron Microscopic images. The antimicrobial activity of MPP against the seven MDR bacteria remained stable after two years of cold storage at 8-10 °C as contrasted to Gentamycin, which lost 20-72% of its antimicrobial effectiveness.

The efficacy of some probiotics and prebiotics on the prevalence of E. coli and the immune response of chickens.

The present study aimed to investigate the efficacy of probiotics and prebiotics in controlling Escherichia coli (E. coli) spp. isolated from chicken. A total of 230 birds representing 19 different commercial breeds were taken from various points. Birds were monitored for postmortem and clinical investigation. Aseptically collected liver samples, lungs, kidneys, hearts, and yolk sacs were then subjected to bacterial isolation and identification. E. coli were observed in 9 pooled samples from 120 examined with an incidence of 7.5%. Nine farms were E. coli-positive, with an incidence of farm infection of 47.3%. The 9 suspected isolates of E. coli were profiled by morphological and microbiological identification of the colony, motility, and gram reaction. The serogroup analysis showed 9 different E. coli for which 3 other groups were identified: 2 E. coli O78, 3 E. coli O111, and 4 untyped groups. Nine isolates of E. coli were subjected to PCR. Molecular detection of 9 strains was conducted to find the virulence genes of E. coli strains (8 STX1, 4 STX2, and 9 EAE). Probiotics and prebiotics significantly increased the total erythrocytic and leukocytic counts throughout the experiment. The phagocytic percentage's main values at 14 d were 47 and 30%, respectively. An increase in the humoral immunity against Newcastle disease (ND) was noticed after ND vaccination. The geometric mean (HI) was 5.9 and 4.2 for probiotic and prebiotic, respectively. It could be concluded that probiotics and prebiotics could stimulate a nonspecific immune response against experimental infection with a virulent strain of E. coli spp.

Hyaluronic acid production by Klebsiella pneumoniae strain H15 (OP354286) under different fermentation conditions.

BACKGROUND: Hyaluronic acid (HA) has gained significant attention due to its unique physical, chemical, and biological properties, making it widely used in various industries. This study aimed to screen bacterial isolates for HA production, characterize favorable fermentation conditions, and evaluate the inhibitory effect of bacterial HA on cancer cell lines. RESULTS: A total of 108 bacterial isolates from diverse sources were screened for HA production using HPLC, turbidimetric, and carbazole determination methods. Among the HA-producing isolates, Klebsiella pneumoniae H15 isolated from an animal feces sample, was superior in HA production. The strain was characterized based on its morphological, cultural, and biochemical characteristics. Molecular identification using 16S rDNA sequencing and phylogenetic analysis confirmed its identity. Fermentation conditions, including pH, temperature, time, and agitation rate, were optimized to maximize HA production. The basal medium, comprising sucrose (7.0%) as carbon source and combined yeast extract with peptone (1.25% each) as nitrogen substrate, favored the highest HA production at pH 8.0, for 30 h, at 30 °C, under shaking at 180 rpm. The average maximized HA concentration reached 1.5 g L-1. Furthermore, bacterial HA exhibited a significant inhibitory effect on three cancer cell lines (MCF-7, HepG-2 and HCT), with the lowest concentration ranging from 0.98-3.91 µg mL-1. CONCLUSIONS: K. pneumoniae H15, isolated from animal feces demonstrated promising potential for HA production. The most favorable fermentation conditions led to a high HA production. The inhibitory effect of bacterial HA on cancer cell lines highlights its potential therapeutic applications. These findings contribute to a broader understanding and utilization of HA in various industries and therapeutic applications.

Controlling bacterial biofilm formation by native and methylated lupine 11S globulins.

The antibacterial and anti-biofilm activities of the 11S globulins isolated from lupin seeds (Lupinus termis), and its methylated derivative (M11S), were investigated against seven pathogenic gram-positive and gram-negative bacteria. The MIC of 11S ranged from 0.1 to 4.0 µg/ml against 0.025 to 0.50 µg/ml for M11S, excelling some specific antibiotics. The MICs of M11S were 40-80 times lower than some specific antibiotics against gram-positive bacteria and 2-60 times lower than some specific antibiotics against gram-negative bacteria. One MIC of 11S and M11S highly reduced the liquid growth of all tested bacteria during 24 h at 37°C. They also inhibited biofilm formation by 80%-86% and 85%-94%, respectively (gram-positive), and 29%-44% and 43%-50%, respectively (gram-negative). M11S prevented biofilm formation by gram-positive bacteria at minimum biofilm inhibitory concentration (MBIC), 0.025-0.1 µg/ml against 0.1-0.5 µg/ml for gram-negative bacteria, i.e., 4-20 times and 4-7 times anti-biofilm inhibitory action compared with 11S, respectively. Biofilm formation of two bacteria revealed no adhered cells on glass slides for 24 h at 37°C, i.e., was entirely prevented by one MBIC of 11S and M11S. Scanning electron microscopy indicated microbial biofilm deformation under the action of 11S and M11S, indicating their broad specificity and cell membrane-targeted action.

Inhibitory action of three lactic acid bacteria cultures on some food-borne pathogens during pickling of green olive fruits.

The healthy olive pickles are those ones which are made by using lactic acid bacteria (LAB) as starter and protective cultures. Three lactic acid bacteria (LAB) strains namely: Lactobacillus plantarum LPS10 (L. plantarum), Lactobacillus fermentum PP17 (L. fermentum) and Pediococcus acidilactici MH512904 (P. acidilactici) were used for inhibition of some food-borne bacterial pathogens such as Listeria monocytogenes LMG10470 (L. monocytogenes), Staphylococcus aureus ATCC25923 (S. aureus), Bacillus cereus ATCC14579 (B. cereus) and Escherichia coli ATCC25922 (E. coli) in Brain Heart Infusion Broth (BHIB) and during pickling of green olive fruits. Cell free supernatants (CFS) from LAB showed distinctive inhibition of the indicator bacterial pathogens used and the inhibitory activity was more pronounced against Gram positive bacteria than that found against Gram negative E. coli strain used; the inhibitory activity of CFS was more pronounced than that obtained by neutralized cell free supernatants (NCFS). Cultures of LAB were used for inhibition of the food-borne pathogens during pickling of green olive fruits. The food-borne bacterial cells grew in olive pickles brine (control) and their growth (CFU/mL) decreased in treated pickles samples and distinctive differences in growth values (CFU/mL) were observed between control and treated samples. CFS from L. plantarum affected target cells of both L. monocytogenes and E. coli and caused cell deformations, cell shrinkage and cell lysis as showed by Transmission Electron Microscopic (TEM) examinations.

The Healing Capability of Clove Flower Extract (CFE) in Streptozotocin-Induced (STZ-Induced) Diabetic Rat Wounds Infected with Multidrug Resistant Bacteria.

Treatment of diabetic foot ulcer (DFU) is of great challenge as it is shown to be infected by multidrug resistant bacteria (MDR bacteria). Sixty four bacterial isolates were isolated from DFU cases; antibiotic susceptibility tests were carried out for all of them. One bacterial isolate (number 11) was shown to resist the action of 8 out of 12 antibiotics used and was identified by both a Vitek-2 system and 16S rRNA fingerprints as belonging to Proteus mirabilis, and was designated Proteus mirabilis LC587231 (P. mirabilis). Clove flower extract (CFE) inhibited distinctively the P. mirabilis bacterium obtained. GC-MS spectroscopy showed that this CFE contained nine bioactive compounds. The effect of CFE on wound healing of Type 1 diabetic albino rats (Rattus norvegicus) was studied. The results indicated that topical application of CFE hydrogel improved wound size, wound index, mRNA expression of the wound healing markers (Coli1, MMP9, Fibronectin, PCNA, and TGFß), growth factor signaling pathways (PPAR-α, PGC1-α, GLP-1, GLPr-1, EGF-ß, EGF-ßr, VEGF-ß, and FGF-ß), inflammatory cytokine expression (IL8, TNFα, NFKß, IL1ß, and MCP1), as well as anti-inflammatory cytokines (IL4 & IL10), pro-apoptotic markers (FAS, FAS-L, BAX, BAX/BCL-2, Caspase-3, P53, P38), as well as an antiapoptotic one (BCL2). Furthermore, it improved the wound oxidative state and reduced the wound microbial load, as the cefepime therapy improved the wound healing parameters. Based on the previous notions, it could be concluded that CFE represents a valid antibiotics alternative for DFU therapy since it improves diabetic wound healing and exerts antibacterial activity either in vitro or in vivo.

Isolation and Characterization of Antibacterial Conglutinins from Lupine Seeds.

The main target of this work is to discover new protein fractions from natural resources with high antibacterial action. The 7S and 11S globulin fractions, as well as the basic subunit (BS), were isolated from lupine seeds (Lupinus termis), chemically characterized, and screened for antibacterial activity against seven pathogenic bacteria. SDS-PAGE revealed molecular weights ranging from 55 to 75 kDa for 7S globulin, 20-37 kD for 11S globulin, and 20 kD for the BS. 11S globulin and the BS migrated faster on Urea-PAGE toward the cathode compared to 7S globulin. FTIR and NMR showed different spectral patterns between the 7S and 11S globulins but similar ones between 11S globulin and the BS. The MICs of the BS were in the range of 0.05-2 µg/mL against Listeria monocytogenes, Klebsiella oxytoca, Proteus mirabilis, Staphylococcus aureus, Listeria ivanovii, Salmonella typhimurium, and Pseudomonas aeruginosa compared to higher values for 11S globulin. The BS surpassed 11S globulin in antibacterial action, while 7S globulin showed no effect. The MICs of 11S globulin and the BS represented only 5% and 2.5% of the specific antibiotic against L. monocytogenes, respectively. Scanning electron microscopy (SEM) demonstrated different signs of cellular deformation and decay in the protein-treated bacteria, probably due to interaction with the bacterial cell wall and membranes. 11S globulin and the BS can be nominated as effective food biopreservatives.

Silver-Protein Nanocomposites as Antimicrobial Agents.

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.

Antibacterial Peptides Produced by Alcalase from Cowpea Seed Proteins.

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.

Chemical Constitution and Antimicrobial Activity of Kombucha Fermented Beverage.

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.

Chemical Constitution and Antimicrobial Activity of Kefir Fermented Beverage.

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.

Methicillin- and Vancomycin-Resistant Staphylococcus aureus From Humans and Ready-To-Eat Meat: Characterization of Antimicrobial Resistance and Biofilm Formation Ability.

Methicillin-resistant and vancomycin-resistant Staphylococcus aureus (MRSA and VRSA) are zoonotic life-threatening pathogens, and their presence in food raises a public health concern. Yet, scarce data are available regarding MRSA and VRSA in both ready-to-eat (RTE) meat and food handlers. This study was undertaken to determine the frequency, antimicrobial resistance, and biofilm-forming ability of MRSA and VRSA isolated from RTE meat (shawarma and burger) and humans (food handlers, and hospitalized patients) in Zagazig city, Sharkia Governorate, Egypt. We analyzed 176 samples (112 human samples: 72 from hospitalized patients and 40 from food handlers, 64 RTE meat samples: 38 from shawarma and 26 from burger). Using phenotypic, PCR-based identification of nuc gene and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), 60 coagulase-positive S. aureus (COPS) isolates were identified in the samples as follow: RTE meat (15/64, 23.4%), hospitalized patients (33/72, 45.8%) and food handlers (12/40, 30%). All the COPS isolates were mecA positive (and thus were classified as MRSA) and multidrug resistant with multiple antibiotic resistance indices ranging from 0.25 to 0.92. Overall, resistance to cefepime (96.7%), penicillin (88.3%), were common, followed by ampicillin-sulbactam (65%), ciprofloxacin (55%), nitrofurontoin (51.7%), and gentamicin (43.3%). VRSA was detected in 30.3% of COPS hospitalized patient's isolates, 26.7% of COPS RTE meat isolates and 25% of COPS food handler's isolates. VanA, vanB, or both genes were detected in 64.7, 5.9, and 29.4% of all VAN-resistant isolates, respectively. The majority of the COPS isolates (50/60, 83.3%) have biofilm formation ability and harbored icaA (76%), icaD (74%), icaC (50%), and icaB (46%) biofilm-forming genes. The bap gene was not detected in any of the isolates. The ability of MRSA and VRSA isolates to produce biofilms in addition to being resistant to antimicrobials highlight the danger posed by these potentially virulent microorganisms persisting in RTE meat, food handlers, and patients. Taken together, good hygiene practices and antimicrobial surveillance plans should be strictly implemented along the food chain to reduce the risk of colonization and dissemination of MRSA and VRSA biofilm-producing strains.

Powerful Antibacterial Peptides from Egg Albumin Hydrolysates.

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.

Inhibition of Staphylococcus aureus LC554891 by Moringa oleifera Seed Extract either Singly or in Combination with Antibiotics.

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.

Detection, Purification and Elucidation of Chemical Structure and Antiproliferative Activity of Taxol Produced by Penicillium chrysogenum.

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.

Catfish Glycoprotein, a Highly Powerful Safe Preservative of Minced Beef Stored at 4 °C for 15 Days.

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.

Exploiting the Biosynthetic Potency of Taxol from Fungal Endophytes of Conifers Plants; Genome Mining and Metabolic Manipulation.

Endophytic fungi have been considered as a repertoire for bioactive secondary metabolites with potential application in medicine, agriculture and food industry. The biosynthetic pathways by fungal endophytes raise the argument of acquisition of these machineries of such complex metabolites from the plant host. Diterpenoids "Taxol" is the most effective anticancer drug with highest annual sale, since its discovery in 1970 from the Pacific yew tree, Taxus brevifolia. However, the lower yield of Taxol from this natural source (bark of T. brevifolia), availability and vulnerability of this plant to unpredicted fluctuation with the ecological and environmental conditions are the challenges. Endophytic fungi from Taxus spp. opened a new avenue for industrial Taxol production due to their fast growth, cost effectiveness, independence on climatic changes, feasibility of genetic manipulation. However, the anticipation of endophytic fungi for industrial Taxol production has been challenged by the loss of its productivity, due to the metabolic reprograming of cells, downregulating the expression of its encoding genes with subculturing and storage. Thus, the objectives of this review were to (1) Nominate the endophytic fungal isolates with the Taxol producing potency from Taxaceae and Podocarpaceae; (2) Emphasize the different approaches such as molecular manipulation, cultural optimization, co-cultivation for enhancing the Taxol productivities; (3) Accentuate the genome mining of the rate-limiting enzymes for rapid screening the Taxol biosynthetic machinery; (4) Triggering the silenced rate-limiting genes and transcriptional factors to activates the biosynthetic gene cluster of Taxol.

Identification and Testing of Antidermatophytic Oxaborole-6-Benzene Sulphonamide Derivative (OXBS) from Streptomyces atrovirens KM192347 Isolated from Soil.

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. .

Antimicrobial Activity and Chemical Constitution of the Crude, Phenolic-Rich Extracts of Hibiscus sabdariffa, Brassica oleracea and Beta vulgaris.

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.

Biological Characterization and Inhibition of Streptococcus pyogenes ZUH1 Causing Chronic Cystitis by Crocus sativus Methanol Extract, Bee Honey Alone or in Combination with Antibiotics: An In Vitro Study.

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.