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1.
Int Microbiol ; 2024 Mar 27.
Artículo en Inglés | MEDLINE | ID: mdl-38532184

RESUMEN

Drug repurposing constitutes a strategy to combat antimicrobial resistance, by using agents with known safety, pharmacokinetics, and pharmacodynamics. Previous studies have implemented new fusidic acid (FA) front-loading-dose regimens, allowing higher serum levels than those achievable with ordinary doses. As susceptibility breakpoints are affected by serum level, we evaluated the repurposing of FA as an antimicrobial product against enterococci. FA minimum inhibitory concentrations (MICs) against standard enterococci strains; Enterococcus faecalis ATCC 29212 and Enterococcus faecium ATCC 27270 were 2 and 4 µg/mL, respectively. The MIC against 98 enterococcal clinical isolates was ≤ 8 µg/mL; all would be susceptible if categorized according to recalculated breakpoints (≥ 16 µg/mL), based on the serum level achieved using the front-loading regimen. FA administration in vivo, using the BALB/c mouse infection model, significantly reduced bacterial burden by two to three log10 units in the liver and spleen of mice infected with vancomycin-susceptible and -resistant strains. Exposure of the standard enterococcal strains to increasing, but not fixed, FA concentrations resulted in resistant strains (MIC = 128 µg/mL), with thicker cell walls and slower growth rates. Only one mutation (M651I) was detected in the fusA gene of the resistant strain derived from serial passage of E. faecium ATCC 27270, which was retained in the revertant strain after passage in the FA-free medium. In conclusion, FA can be repurposed as an antimicrobial drug against enterococci with a low probability of mutational resistance development, and can be employed for treatment of infections attributable to vancomycin-resistant enterococci.

2.
Curr Microbiol ; 74(12): 1365-1372, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-28803266

RESUMEN

A clean way to overcome environmental pollution is biodegradation. In this perspective, at the intersection of biodegradation and metagenomics, the degradome is defined as the totality of genes related to the biodegradation of a certain compound. It includes the genetic elements from both culturable and uncultured microorganisms. The possibility of assessing the biodegradation potential of an environmental samples, using a degradome-based polymerase chain reaction, was explored. 2,4-Dichlorophenol (2,4-DCP) was chosen as a model and the use of tfdB gene as a biodegradation marker was confirmed by bioinformatics study of TfdB protein. Five primer pairs were designed for the detection of different tfdB gene families. A total of 16 environmental samples were collected from Egyptian agricultural soils and wastewaters and tested for the presence of 2,4-DCP. The biodegradation capacity of 2,4-DCP was determined, for all isolated consortia, to reach up to 350 mg/l. Metagenomic DNA was extracted directly from the soil samples while successive 2,4-DCP-degrading microbial communities were enriched, with increasing concentrations of 2,4-DCP, then their DNA was extracted. The extracted DNA was tested for the distribution of the tfdB gene using a degradome-based polymerase chain reaction. tfdB-1 and tfdB-2 were detected in 5 and 9 samples, respectively. However, the co-existence of both genes was detected only in five samples. All tfdB positive samples were capable of 2,4-DCP degradation. The developed approach of assessing the potential of different environments for degrading 2,4-DCP was successfully measured in terms of accuracy (81.25%) and specificity (100%).


Asunto(s)
Clorofenoles/metabolismo , Contaminantes Ambientales/metabolismo , Redes y Vías Metabólicas , Oxigenasas de Función Mixta/análisis , Reacción en Cadena de la Polimerasa/métodos , Microbiología del Suelo , Biotransformación , Egipto , Metagenómica/métodos , Oxigenasas de Función Mixta/genética
3.
Biotechnol Lett ; 38(9): 1493-502, 2016 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-27271522

RESUMEN

OBJECTIVE: To test the toxicity of ketoprofen (a commonly-used NSAIDs) using two microalgal strains and Artemia sp. following the isolation of bacterial and microalgal strains and testing their ability to biodegrade and tolerate ketoprofen. RESULTS: Chlorella sp. was the most resistant to ketoprofen. A defined bacterial consortium (K2) degraded 5 mM ketoprofen as a sole carbon source both in the dark or continuous illumination. Ketoprofen did not undergo photodegradation. In the dark, biodegradation was faster with a lag phase of 10 h, 41% COD removal and 82 % reduction in toxicity. The consortium degraded up to 16 mM ketoprofen. The consortium was composed of four bacterial isolates that were identified. MS/MS analysis suggested a ketoprofen biodegradation pathway that has not been previously reported. Combining Chlorella sp. and the K2 consortium, ketoprofen was degraded within 7 days under a diurnal cycle of 12 h light/12 h dark. CONCLUSION: The feasibility of using a microalgal-bacterial system to treat pharmaceutical wastewater is promising for the reduction of the process cost and providing a safer technology for pharmaceutical wastewater treatment.


Asunto(s)
Bacterias/metabolismo , Cetoprofeno/farmacología , Microalgas/metabolismo , Bacterias/efectos de los fármacos , Microalgas/efectos de los fármacos , Fotoquímica , Spirulina/efectos de los fármacos , Spirulina/metabolismo , Eliminación de Residuos Líquidos
4.
Drug Dev Ind Pharm ; 42(11): 1752-62, 2016 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-27093938

RESUMEN

CONTEXT: A microbiological multidistrict-based survey from different Egyptian governorates was conducted to determine the most prevalent causative agents of ocular infections in the Egyptian population. Antibiotic sensitivity testing was then performed to identify the most potent antimicrobial agent. Vancomycin (VCM) proved the highest activity against gram-positive Staphylococcus bacteria, which are the most commonly isolated causative agents of ocular infection. However, topically applied VCM suffers from poor ocular bioavailability because of its high molecular weight and hydrophilicity. OBJECTIVE: The aim of the present study was to develop VCM-loaded solid lipid nanoparticles (SLNs) using water-in-oil-in-water (W/O/W) double emulsion, solvent evaporation technique to enhance ocular penetration and prolong ophthalmic residence of VCM. METHOD: Two consecutive full factorial designs (2(4) followed by 3(2)) were adopted to study the effect of different formulation and process parameters on SLN formulation. The lipid type and structure, polyvinyl alcohol (PVA) molecular weight and concentration, sonication time, as well as lipid:drug ratio were studied as independent variables. The formulated SLN formulae were evaluated for encapsulation efficiency (EE%), particle size (PS), and zeta potential as dependent variables. RESULTS: The statistically-optimized SLN formula (1:1 ratio of glyceryltripalmitate:VCM with 1% low molecular weight PVA and 1 min sonication time) had average PS of 277.25 nm, zeta potential of -20.45, and 19.99% drug encapsulation. Scanning and transmission electron micrographs showed well-defined, spherical, homogenously distributed particles. CONCLUSION: The present study suggests that VCM incorporation into SLNs is successfully achievable; however, further studies with different nanoencapsulation materials and techniques would be valuable for improving VCM encapsulation.


Asunto(s)
Sistemas de Liberación de Medicamentos/métodos , Emulsiones/química , Ojo/microbiología , Lípidos/química , Nanopartículas/química , Alcohol Polivinílico/química , Vancomicina/administración & dosificación , Vancomicina/farmacología , Ojo/química , Interacciones Hidrofóbicas e Hidrofílicas , Fenómenos Microbiológicos , Tamaño de la Partícula , Sonicación , Vancomicina/química
5.
Biotechnol Lett ; 36(9): 1773-81, 2014 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-24930096

RESUMEN

The effect of inorganic pollutants on the treatment of organic pollutants using algal/bacterial microcosm was investigated in a continuous photobioreactor. The microcosm was composed of Chlorella vulgaris MM1 and Pseudomonas MT1 and was able to efficiently treat artificial waste-water contaminated with 6.4 salicylate and 2.2 mM phenol at a hydraulic retention time of 4 days. No negative effect was recorded when the waste-water was supplemented with 1.6 mM thiocyanate; however, the treatment efficiency severely deteriorated when the system was challenged with 0.74 mM cyanide. Addition of 2 g NaHCO3 l(-1) did not improve the efficiency of the treatment. Toxicity of the pollutants to the alga was cyanide > thiocyanate > phenol > salicylate. The high toxicity of the waste-water was eliminated either by a 25-fold dilution or by photocatalytic pre-treatment which allowed the subsequent efficient biological treatment.


Asunto(s)
Chlorella vulgaris/efectos de los fármacos , Cianuros/metabolismo , Cianuros/toxicidad , Compuestos Orgánicos/metabolismo , Fotobiorreactores/microbiología , Pseudomonas/efectos de los fármacos , Contaminantes Químicos del Agua/metabolismo , Chlorella vulgaris/crecimiento & desarrollo , Chlorella vulgaris/metabolismo , Pseudomonas/crecimiento & desarrollo , Pseudomonas/metabolismo
6.
ScientificWorldJournal ; 2014: 549858, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-25506614

RESUMEN

Mycoplasmas are fastidious slow growing organisms lacking a cell wall and mostly isolated from the mucosal surfaces of the respiratory and genitourinary tracts. There is a dearth of information regarding clinical Mycoplasma spp. isolates among Egyptian patients. A total of 170 samples were collected from patients and apparently healthy personnel in local public hospitals in Cairo, Egypt. Isolation of Mycoplasma spp. was carried out using appropriate culture media and further identification was carried out by biochemical tests followed by serotyping using specific antisera. Confirmation was done by PCR for detection of different Mycoplasma spp. using genus-specific primers targeting 16S ribosomal RNA gene. Characterization of the antibiotic resistance and sensitivity pattern against different antimicrobials was carried out using disc diffusion test. The results indicated the presence of six Mycoplasma spp. in 22.94% of the samples. Mycoplasmas were detected more frequently in throat swabs than sputum. Mycoplasma pneumoniae was highly sensitive to macrolides and quinolones but less sensitive to aminoglycosides and tetracyclines. Molecular techniques were found to be of more rapid, highly sensitive, able to detect nonviable organisms, and cost effective. These results shed light on difficulties of Mycoplasma detection and the superiority of molecular techniques over culture.


Asunto(s)
Técnicas de Tipificación Bacteriana/métodos , Hospitales , Tipificación Molecular/métodos , Mycoplasma/clasificación , Mycoplasma/aislamiento & purificación , Recuento de Colonia Microbiana , Egipto , Humanos , Datos de Secuencia Molecular , Mycoplasma/crecimiento & desarrollo
7.
World J Microbiol Biotechnol ; 29(6): 969-74, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23296918

RESUMEN

An algal-bacterial microcosm was synthetically constructed of Chlorella vulgaris MMl and Pseudomonas MTl. This microcosm was able to treat simulated wastewater supplemented with mixtures of phenol and pyridine up to 4.6 and 4.4 mM, respectively, in a continuous stirred tank bioreactor (CSTR) using photosynthetic oxygenation. Complete pollutant removal and detoxification and 82 % removal of introduced chemical oxygen demand (COD) were achieved at a hydraulic retention time (HRT) of 2.7 days. Increasing the influent load to 5.3 and 6.3 mM reduced the removal of phenol, pyridine and COD to 78, 21 and 59 %, respectively. Fertilization of the photobioreactor with 24 mM NaHCO3 restored the treatment and detoxification efficiencies. The system was able to additionally mitigate up to 72 mM NaHCO3 at the same HRT. Although the fertilization increased the system treatment efficiency, the settleability of the algal-bacterial microcosm was significantly reduced. When the photobioreactor was operated at HRT of 2.7 days in a 12/12 h of dark/light cycle, complete removal of 4.7 mM phenol was recorded but only 11 % of 5.7 mM pyridine was removed. The COD removal efficiency and CO2 mitigation were also reduced to 65 and 86 %, respectively, and the effluent retained significant toxicity where 73 % inhibition was recorded. Elongation of the illumination time to 48 h (HRT of 4 days at 12/12 h dark/light cycle) restored the treatment and detoxification efficiencies.


Asunto(s)
Dióxido de Carbono/metabolismo , Chlorella vulgaris/metabolismo , Fotobiorreactores/microbiología , Pseudomonas/metabolismo , Contaminantes Químicos del Agua/metabolismo , Biotransformación , Datos de Secuencia Molecular , Oxidación-Reducción , Fenol/metabolismo , Fotosíntesis , Piridinas/metabolismo , Análisis de Secuencia de ADN
8.
Lancet Gastroenterol Hepatol ; 7(7): 658-665, 2022 07.
Artículo en Inglés | MEDLINE | ID: mdl-35489364

RESUMEN

Until 2018, Egypt had the highest prevalence of hepatitis C virus (HCV) infection globally, affecting approximately 7% of the population. Despite efforts in diagnosis and treatment since 2006, nearly 2 million individuals with chronic HCV infection had yet to be diagnosed as of early 2018. In December, 2018, a mass HCV screening campaign for adolescents aged 15-18 years was initiated. Among 3 024 325 adolescents screened, the HCV antibody seroprevalence was 11 477 (0·38%), of whom 8187 (78·7%) were HCV RNA-positive. Sustained virological response 12 weeks after completion of treatment (SVR12) was attained by 7327 (99·6%) adolescents with a fixed-dose combination of generic ledipasvir 90 mg plus sofosbuvir 400 mg. Although mass screening in this age group might not be regularly adopted by many health systems and its cost-effectiveness might be lower than the screening of adults and high-risk groups (eg, patients on haemodialysis, people who inject drugs), breaking the chain of transmission in younger populations should lead to a reduction in HCV incidence and complications, and hasten the elimination of the disease.


Asunto(s)
Hepacivirus , Hepatitis C Crónica , Adolescente , Adulto , Antivirales/uso terapéutico , Egipto/epidemiología , Genotipo , Hepacivirus/genética , Hepatitis C Crónica/diagnóstico , Hepatitis C Crónica/tratamiento farmacológico , Hepatitis C Crónica/epidemiología , Humanos , Tamizaje Masivo , Instituciones Académicas , Estudios Seroepidemiológicos
9.
Drug Dev Ind Pharm ; 37(12): 1491-508, 2011 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-21707323

RESUMEN

Nonionic surfactant (NIS) vesicles (niosomes) formed from self-assembly of hydrated synthetic NIS monomers are capable of entrapping a variety of drugs and have been evaluated as an alternative to liposomes. Nystatin (NYS) is a polyene antifungal drug that has been used in the treatment of cutaneous, vaginal and oral fungal infections since the 1950s. The aim of this work is to encapsulate NYS in niosomes to obtain a safe and effective formula administered parenterally for neutropenic patients. NYS niosomes were prepared by the thin-film hydration method using Span 60 or Span 40 and cholesterol (CHOL). Stearylamine and dicetyl phosphate were added as the positive and negative charge-inducing agents (CIA), respectively. Two molar ratios were used, namely NIS/CHOL/CIA (1:1:0.1 and 2:1:0.25). Neutral and positively charged niosomes gave the highest encapsulation efficiencies. NYS niosomes were characterized using transmission electron microscopy, differential scanning calorimetry and dynamic light scattering. The release of neutral and negatively charged NYS niosomes was estimated, and it showed a slow sustained release profile. A 25-kGy γ-irradiation dose was sufficient to sterilize the investigated vesicles. NYS niosomes exerted less nephrotoxicity and hepatotoxicity in vivo, showed higher level of drug in vital organs and revealed pronounced efficacy in elimination of the fungal burden in experimental animals infected with Candida albicans compared with those treated with free NYS. Niosomal encapsulation thus provided means for parenteral administration of NYS, reducing its toxicity and making it a more active antifungal agent.


Asunto(s)
Antifúngicos/administración & dosificación , Candidiasis/tratamiento farmacológico , Sistemas de Liberación de Medicamentos , Liposomas/administración & dosificación , Nistatina/administración & dosificación , Análisis de Varianza , Animales , Antifúngicos/química , Antifúngicos/farmacología , Modelos Animales de Enfermedad , Femenino , Liposomas/química , Liposomas/farmacología , Masculino , Ratones , Nistatina/química , Nistatina/farmacología , Distribución Aleatoria , Ratas , Ratas Wistar
10.
Toxicon ; 195: 7-16, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33610638

RESUMEN

Aflatoxins are carcinogenic compounds produced by certain Aspergillus spp and naturally contaminate poultry rations. Exposure to low levels of Aflatoxin B1 (AFB1) in poultry feeds is the second most threatening issue facing the poultry industry in Egypt; it can cause a reduction in growth, egg production, and compromised immune functions, resulting in significant economic loss. Hence, a safe, effective and eco-friendly detoxification method is strongly required. Biological decontamination is a promising approach to reduce aflatoxin levels within threshold limits. This study explores the biodegradation capacity of bacteria isolated from the moldy feed, soil and poultry feces in various poultry farms against AFB1 (100 ppb), G1 (100 ppb), B2 (30 ppb), G2 (30 ppb). Sixty-five bacterial isolates were initially screened using coumarin media with a concentration of (0.01%-0.5%) coumarin. Only one soil isolate (SZ1) grew at the highest concentration (0.5%). Coumarin and Aflatoxin degradation rates of ten promising isolates were measured using spectrophotometry and HPLC. Six isolates reduced AFG1 by more than 90% in the liquid medium, five reduced AFB2 while only four did the same with AFB1& AFG2. Impressively, isolate SZ1 (identified as Pseudomonas fluorescens) exhibited the best degradation capacity to both coumarin and aflatoxin with 100% degradation of AFG1 and 99% degradation of AFB1, AFB2 and AFG2. Biochemical and molecular identification of the ten isolates revealed that they belong to four genera; Bacillus (6), Pseudomonas (2), Enterococcus (1) and Stenotrophomonas (1). Factors affecting Pseudomonas fluorescens SZ1 degradation activity was further investigated. Optimum temperature, time and pH for maximum aflatoxin degradation were at 37 °C, 72 h and 7, respectively. Treatment with proteinase K reduced the degradation activity of G1 (31% ± 1.438), B1 (42% ± 1.438), G2 (19% ± 1.097), and B2 (25% ± 1.732), suggesting that the effective component in aflatoxin degradation may be protein in nature. Our study suggests the biocontrol potential of several different species isolated from poultry farms; B. haynesii, B. licheniformis, B. tequilensis, B. subtilis, B. amyloliquefaciens, Pseudomonas fluorescens, Enterococcus casseliflavus, and Stenotrophomonas maltophilia. The results proposed Pseudomonas fluorescens SZ1 as an excellent candidate for bioremediation and decontamination of aflatoxin in feed matrices. To the best of our knowledge, this is the first report identifying B. haynesii, Enterococcus casseliflavus, B. tequilensis and B. amyloliquefaciens with aflatoxin degradation activity.


Asunto(s)
Aflatoxinas/metabolismo , Biodegradación Ambiental , Aflatoxina B1/análisis , Aflatoxinas/análisis , Animales , Egipto , Enterococcus , Granjas , Aves de Corral
11.
Sci Rep ; 11(1): 3795, 2021 02 15.
Artículo en Inglés | MEDLINE | ID: mdl-33589735

RESUMEN

Nosocomial infections caused by enterococci are an ongoing global threat. Thus, finding therapeutic agents for the treatment of such infections are crucial. Some Enterococcus faecalis strains are able to produce antimicrobial peptides called bacteriocins. We analyzed 65 E. faecalis isolates from 43 food samples and 22 clinical samples in Egypt for 17 common bacteriocin-encoding genes of Enterococcus spp. These genes were absent in 11 isolates that showed antimicrobial activity putatively due to bacteriocins (three from food, including isolate OS13, and eight from clinical isolates). The food-isolated E. faecalis OS13 produced bacteriocin-like inhibitory substances (BLIS) named enterocin OS13, which comprised two peptides (enterocin OS13α OS13ß) that inhibited the growth of antibiotic-resistant nosocomial E. faecalis and E. faecium isolates. The molecular weights of enterocin OS13α and OS13ß were determined as 8079 Da and 7859 Da, respectively, and both were heat-labile. Enterocin OS13α was sensitive to proteinase K, while enterocin OS13ß was resistant. Characterization of E. faecalis OS13 isolate revealed that it belonged to sequence type 116. It was non-hemolytic, bile salt hydrolase-negative, gelatinase-positive, and sensitive to ampicillin, penicillin, vancomycin, erythromycin, kanamycin, and gentamicin. In conclusion, BLIS as enterocin OS13α and OS13ß represent antimicrobial agents with activities against antibiotic-resistant enterococcal isolates.


Asunto(s)
Bacteriocinas/farmacología , Infección Hospitalaria/tratamiento farmacológico , Farmacorresistencia Bacteriana/efectos de los fármacos , Enterococcus faecalis/química , Bacteriocinas/química , Bacteriocinas/aislamiento & purificación , Infección Hospitalaria/microbiología , Farmacorresistencia Bacteriana/genética , Egipto , Enterococcus faecalis/efectos de los fármacos , Enterococcus faecalis/genética , Enterococcus faecalis/patogenicidad , Enterococcus faecium/efectos de los fármacos , Enterococcus faecium/patogenicidad , Microbiología de Alimentos , Humanos , Pruebas de Sensibilidad Microbiana
12.
Sci Total Environ ; 677: 120-130, 2019 Aug 10.
Artículo en Inglés | MEDLINE | ID: mdl-31055095

RESUMEN

Nowadays, due to worldwide water shortage, water utilities are forced to re-evaluate treated wastewater. Consequently, wastewater treatment plants need to conduct biomonitoring. Coking wastewater (CWW) has toxic, mutative and carcinogenic components with threatening effect on the environment. CWW was selected as a model for complex highly toxic industrial wastewater that should be treated. CWW from Egypt was treated in a nine-liter photobioreactor using an algal-bacterial system. The photobioreactor was operated for 154 days changing different parameters (toxic load and light duration) for optimization. Optimized conditions achieved significant reduction (45%) in the operation cost. The algal-bacterial system was monitored using chemical assays (chemical oxygen demand and phenol analysis), bioassays (phytotoxicity, Artemia-toxicity, cytotoxicity, algal-bacterial ratio and settleability) and Illumina-MiSeq sequencing of 16S rRNA gene. The algal-bacterial system detoxified (in terms of phytotoxicity, cytotoxicity and Artemia-toxicity) CWW introduced as influent through all phases. A significant difference was recorded in the microbial diversity between influent and effluent samples. Four phyla dominated influent samples; Proteobacteria (77%), Firmicutes (11%), Bacteroidetes (5%) and Deferribacteres (3%) compared to only two in effluent samples; Proteobacteria (66%) and Bacteroidetes (26%). The significant relative-abundance of versatile aromatic degraders (Comamonadaceae and Pseudomonadaceae families) in influent samples conformed to the nature of CWW. Microbial community shifted and promoted the activity of catabolically versatile and xenobiotics degrading families (Chitinophagaceae and Xanthomonadaceae). Co-culture of microalgae had a positive effect on the biodegrading bacteria that was reflected by enhanced treatment efficiency, significant increase in relative abundance of bacterial genera with cyanide-decomposing potential and negative effect on waterborne pathogens.


Asunto(s)
Bacterias/metabolismo , Chlorella vulgaris/metabolismo , Monitoreo del Ambiente/métodos , Restauración y Remediación Ambiental/métodos , Aguas Residuales/análisis , Aguas Residuales/microbiología , Coque , Egipto , Microalgas/metabolismo , Microbiota , Contaminantes Químicos del Agua/análisis , Contaminación Química del Agua/prevención & control
13.
Biomolecules ; 10(1)2019 12 27.
Artículo en Inglés | MEDLINE | ID: mdl-31892223

RESUMEN

Mycobacteriophage endolysins have emerged as a potential alternative to the current antimycobacterial agents. This study focuses on mycolylarabinogalactan hydrolase (LysB) enzymes of the α/ß-hydrolase family, which disrupt the unique mycolic acid layer of mycobacterium cell wall. Multiple sequence alignment and structural analysis studies showed LysB-D29, the only enzyme with a solved three-dimensional structure, to share several common features with esterases (lacking lid domain) and lipases (acting on long chain lipids). Sequence and structural comparisons of 30 LysB homology models showed great variation in domain organizations and total protein length with major differences in the loop-5 motif harboring the catalytic histidine residue. Docking of different p-nitrophenyl ligands (C4-C18) to LysB-3D models revealed that the differences in length and residues of loop-5 contributed towards wide diversity of active site conformations (long tunnels, deep and superficial funnels, shallow bowls, and a narrow buried cave) resembling that of lipases, cutinases, and esterases. A set of seven LysB enzymes were recombinantly produced; their activity against p-nitrophenyl esters could be related to their active site conformation and acyl binding site. LysB-D29 (long tunnel) showed the highest activity with long chain p-nitrophenyl palmitate followed by LysB-Omega (shallow bowl) and LysB-Saal (deep funnel).


Asunto(s)
Esterasas/química , Esterasas/metabolismo , Galactanos/metabolismo , Micobacteriófagos/enzimología , Secuencia de Aminoácidos , Esterasas/genética , Modelos Moleculares , Simulación del Acoplamiento Molecular , Conformación Proteica , Alineación de Secuencia
14.
Braz J Microbiol ; 49 Suppl 1: 1-8, 2018 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-29631893

RESUMEN

In this study, the development and assessment of a modified, efficient, and cost-efficient protocol for mDNA (metagenomic DNA) extraction from contaminated water samples was attempted. The efficiency of the developed protocol was investigated in comparison to a well-established commercial kit (Epicentre, Metagenomic DNA Isolation Kit for Water). The comparison was in terms of degree of shearing, yield, purity, duration, suitability for polymerase chain reaction and next-generation sequencing in addition to the quality of next-generation sequencing data. The DNA yield obtained from the developed protocol was 2.6 folds higher than that of the commercial kit. No significant difference in the alpha (Observed species, Chao1, Simpson and PD whole tree) and beta diversity was found between the DNA samples extracted by the commercial kit and the developed protocol. The number of high-quality sequences of the samples extracted by the developed method was 20% higher than those obtained by the samples processed by the kit. The developed economic protocol successfully yielded high-quality pure mDNA compatible with complex molecular applications. Thus we propose the developed protocol as a gold standard for future metagenomic studies investigating a large number of samples.


Asunto(s)
Métodos Analíticos de la Preparación de la Muestra/métodos , Bacterias/aislamiento & purificación , ADN Bacteriano/aislamiento & purificación , Agua Dulce/microbiología , Metagenómica/economía , Metagenómica/métodos , Métodos Analíticos de la Preparación de la Muestra/economía , Bacterias/clasificación , Bacterias/genética , ADN Bacteriano/genética , Agua Dulce/química , Filogenia , Análisis de Secuencia de ADN
15.
AMB Express ; 8(1): 83, 2018 May 21.
Artículo en Inglés | MEDLINE | ID: mdl-29785517

RESUMEN

Azo dyes are complex derivatives of diazene used in food and textile manufacture. They are highly recalcitrant compounds, and account for severe environmental and health problems. Different strains of Pseudomonas species were isolated from textile wastewater effluents. The bioconversion of Remazol black B (a commonly used water soluble dye) by Pseudomonas aeruginosa was observed in static conditions. The bio-decolorization process was optimized by a multi factorial Plackett-Burman experimental design. Decolorization of 200 mg L-1 reached 100% in 32 h. Interestingly, the presence of yeast extract, magnesium and iron in the culture media, highly accelerated the rate of decolorization. Moreover, one of our isolates, P. aeruginosa KY284155, was kept high degradation rates at high pH (pH = 9), which represents the pH of most textile wastewater effluents, and was able to tolerate high concentration of dye up to 500 mg L-1. In bacteria, azo-dye degradation is often initiated by reductive azo compound cleavage catalyzed by azo-reductases. Three genes encoding azo-reductases, paazoR1, paazoR2 and paazoR3, could be identified in the genome of the isolated P. aeruginosa stain (B1). Bioinformatics analyses of the paazoR1, paazoR2 and paazoR3 genes reveal their prevalence and conservation in other P. aeruginosa strains. Chemical oxygen demand dramatically decreased and phyto-detoxification of the azo dye was accomplished by photocatalytic post treatment of the biodegradation products. We suggest applying combined biological photocatalytic post treatment for azo dyes on large scale, for effective, cheap decolorization and detoxification of azo-dyes, rendering them safe enough to be discharged in the environment.

16.
Water Res ; 41(8): 1697-704, 2007 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-17350074

RESUMEN

Simulated solar UV/TiO(2) photocatalysis was efficient to detoxify a mixture of 100 mgphenoll(-1) and 50 mgp-nitrophenol (PNP) l(-1) and allow the subsequent biodegradation of the remaining pollutants and their photocatalytic products under photosynthetic aeration with Chlorella vulgaris. Photocatalytic degradation of phenol and PNP was well described by pseudo-first order kinetics (r(2)>0.98) with removal rate constants of 1.9x10(-4) and 2.8x10(-4)min(-1), respectively, when the pollutants were provided together and 5.7x10(-4) and 9.7x10(-4)min(-1), respectively, when they were provided individually. Photocatalytic pre-treatment of the mixture during 60 h removed 50+/-1% and 62+/-2% of the phenol and PNP initially present but only 11+/-3% of the initial COD. Hydroquinone, nitrate and catechol were identified as PNP photocatalytic products and catechol and hydroquinone as phenol photocatalytic products. Subsequent biological treatment of the pre-treated samples removed the remaining contaminants and their photocatalytic products as well as 81-83% of the initial COD, allowing complete detoxification of the mixture to C. vulgaris. Similar detoxification efficiencies were recorded after biological treatment of the irradiated mixture with activated sludge microflora or with an acclimated consortia composed of a phenol-degrading Alcaligenes sp. and a PNP-degrading Arthrobacter sp., although the acclimated strains biodegraded the remaining pollutants faster. Biological treatment of the non-irradiated mixture was inefficient due to C. vulgaris inhibition.


Asunto(s)
Nitrofenoles , Fenol , Eliminación de Residuos Líquidos/métodos , Contaminantes Químicos del Agua , Alcaligenes/metabolismo , Arthrobacter/metabolismo , Catálisis , Chlorella vulgaris/efectos de los fármacos , Chlorella vulgaris/metabolismo , Clorofila/metabolismo , Lepidium sativum/efectos de los fármacos , Lepidium sativum/crecimiento & desarrollo , Nitrofenoles/química , Nitrofenoles/metabolismo , Nitrofenoles/efectos de la radiación , Nitrofenoles/toxicidad , Fenol/química , Fenol/metabolismo , Fenol/efectos de la radiación , Fenol/toxicidad , Fotosíntesis , Tallos de la Planta/efectos de los fármacos , Tallos de la Planta/crecimiento & desarrollo , Titanio/química , Rayos Ultravioleta , Contaminantes Químicos del Agua/química , Contaminantes Químicos del Agua/metabolismo , Contaminantes Químicos del Agua/efectos de la radiación , Contaminantes Químicos del Agua/toxicidad
17.
Chemosphere ; 66(11): 2201-9, 2007 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-17097127

RESUMEN

UV/TiO2/H2O2, UV/TiO2 and UV/H2O2 were compared as pre-treatment processes for the detoxification of mixtures of 4-chlorophenol (4CP), 2,4-dichlorophenol (DCP), 2,4,6-trichlorophenol (TCP) and pentachlorophenol (PCP) prior to their biological treatment. When each chlorophenol was initially supplied at 50 mg l(-1), UV/TiO2/H2O2 treatment supported the highest pollutant removal, COD removal, and dechlorination efficiencies followed by UV/TiO2 and UV/H2O2. The remaining toxicity to Lipedium sativum was similar after all pre-treatments. Chlorophenol photodegradation was always well described by a first order model kinetic (r2>0.94) and the shortest 4CP, DCP, TCP and PCP half-lives of 8.7, 7.1, 4.5 and 3.3 h, respectively, were achieved during UV/TiO2/H2O2 treatment. No pollutant removal was observed in the controls conducted with H2O2 or TiO2 only. Inoculation of all the photochemically pre-treated mixtures with activated sludge microflora was followed by complete removal of the remaining pollutants. Combined UV/TiO2/H2O2-biological supported the highest detoxification, dechlorination (99%) and COD removal (88%) efficiencies. Similar results were achieved when each chlorophenol was supplied at 100 mg l(-1). COD and Cl mass balances indicated UV, UV/H2O2, and UV/TiO2 treatments lead to the formation of recalcitrant photoproducts, some of which were chlorinated.


Asunto(s)
Clorofenoles/química , Contaminantes Ambientales/química , Contaminación Ambiental/prevención & control , Peróxido de Hidrógeno/química , Fotoquímica/métodos , Titanio/química , Rayos Ultravioleta , Bacterias/metabolismo , Biodegradación Ambiental , Clorofenoles/metabolismo , Cromatografía Líquida de Alta Presión , Contaminantes Ambientales/metabolismo , Semivida , Cinética , Aguas del Alcantarillado/microbiología
18.
Appl Biochem Biotechnol ; 183(1): 189-199, 2017 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-28236193

RESUMEN

A halophilic cellulase-producing bacterium was isolated from a sediment sample collected from Lake Qarun (Fayoum Province, Egypt). Molecular identification based on 16S rDNA amplification and sequencing revealed 99% homology with Halobacillus sp. and hence was designated as Halobacillus sp. QLS 31. Medium composition and culture conditions were optimized for enhancing the production of cellulase enzyme using the Plackett-Burman statistical design. Ten variables were evaluated for their influence on cellulase production. Carboxymethyl cellulose (CMC), zinc sulfate (ZnSO4), and inoculum size were found to exert a significant effect on cellulase productivity by Halobacillus sp. QLS 31. The maximum specific activity of cellulase enzyme was 48.08 U/mg. Following the predicted conditions, a 7.5-fold increase in cellulase specific activity (175.47 U/mg) was achieved compared to the basal medium (23.19 U/mg) under the following optimized conditions: temperature (30 °C), fermentation time (2 days ), pH value (9), CMC concentration (1%), inoculum size (1%), yeast extract concentration (0.1%), ammonium sulfate ((NH3)2SO4) concentration (0.1%), sodium chloride (NaCl) concentration (20%), and metal inducers: ZnSO4 (0.1%) and Ca/Mg ratio (0.01%). Thus, the results of this study provide an important basis for more efficient, cheap industrial cellulase production from halophilic Halobacillus sp. QLS 31.


Asunto(s)
Proteínas Bacterianas/biosíntesis , Celulasas/biosíntesis , Halobacillus , Lagos/microbiología , Microbiología del Agua , Proteínas Bacterianas/genética , Celulasas/genética , Egipto , Halobacillus/enzimología , Halobacillus/genética , Halobacillus/aislamiento & purificación
19.
Bioresour Technol ; 232: 364-371, 2017 May.
Artículo en Inglés | MEDLINE | ID: mdl-28254731

RESUMEN

An artificial microalgal-bacterial consortium was used to remediate a mixture of analgesics (ketoprofen, paracetamol and aspirin) in a stirred-tank photobioreactor. A hydraulic retention time (HRT) of 3days supported poor treatment because of the formation of p-aminophenol (paracetamol toxic metabolite). Increasing the HRT to 4days enhanced the bioremediation efficiency. After applying an acclimatization regime, 95% removal of the analgesics mixture, p-aminophenol and COD reduction were achieved. However, shortening the HRT again to 3days neither improved the COD reduction nor ketoprofen removal. Applying continuous illumination achieved the best analgesics removal results. The harvested biomass contained 50% protein, which included almost all essential amino acids. The detected fatty acid profile suggested the harvested biomass to be a good biodiesel-producing candidate. The water-extractable fraction possessed the highest phenolic content and antioxidant capacity. These findings suggest the whole process to be an integrated eco-friendly and cost-efficient strategy for remediating pharmaceutical wastewater.


Asunto(s)
Bacterias/metabolismo , Biomasa , Microalgas/metabolismo , Consorcios Microbianos , Fotobiorreactores/microbiología , Acetaminofén/aislamiento & purificación , Aminoácidos/análisis , Analgésicos/aislamiento & purificación , Aspirina/aislamiento & purificación , Técnicas de Cultivo Celular por Lotes , Biodegradación Ambiental , Análisis de la Demanda Biológica de Oxígeno , Clorofila/análisis , Clorofila A , Ácidos Grasos/análisis , Concentración 50 Inhibidora , Preparaciones Farmacéuticas , Pruebas de Toxicidad
20.
Eur J Pharm Sci ; 100: 142-154, 2017 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-28089661

RESUMEN

Ocular topically applied Vancomycin (VCM) suffers poor bioavailability due to its high molecular weight and hydrophilicity. In the present investigation, VCM-loaded polymeric nanoparticles (PNPs) were developed aiming to enhance its ocular bioavailability through prolonging its release pattern and ophthalmic residence. PNPs were prepared utilizing double emulsion (W/O/O), solvent evaporation technique. 23×41 full factorial design was applied to evaluate individual and combined influences of polymer type, Eudragit® RS100, sonication time, and Span®80 concentration on PNPs particle size, encapsulation efficiency, and zeta potential. Further, the optimized formulae were incorporated in 1% Carbopol®-based gel. In-vivo evaluation of the optimized formulae was performed via Draize test followed by microbiological susceptibility testing on albino rabbits. Results revealed successful formulation of VCM-loaded PNPs was achieved with particle sizes reaching 155nm and up to 88% encapsulation. Draize test confirmed the optimized formulae as non-irritating and safe for ophthalmic administration. Microbiological susceptibility testing confirmed prolonged residence, higher Cmax. with more than two folds increment in the AUC(0.25-24) of VCM-PNPs over control groups. Thus, VCM-loaded PNPs represent promising carriers with superior achievements for enhanced Vancomycin ophthalmic delivery over the traditional use of commercially available VCM parenteral powder after constitution into a solution by the ophthalmologists.


Asunto(s)
Antibacterianos , Portadores de Fármacos , Nanopartículas , Vancomicina , Resinas Acrílicas/química , Administración Oftálmica , Animales , Antibacterianos/administración & dosificación , Antibacterianos/química , Antibacterianos/toxicidad , Portadores de Fármacos/administración & dosificación , Portadores de Fármacos/química , Portadores de Fármacos/toxicidad , Composición de Medicamentos , Liberación de Fármacos , Geles , Concentración de Iones de Hidrógeno , Nanopartículas/administración & dosificación , Nanopartículas/química , Nanopartículas/toxicidad , Conejos , Staphylococcus aureus/efectos de los fármacos , Vancomicina/administración & dosificación , Vancomicina/química , Vancomicina/toxicidad
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