A. A. M. Shoreit

Isolation, characterization of heavy metal resistant strain of Pseudomonas aeruginosa isolated from polluted sites in Assiut city, Egypt

Sixty six isolates of Pseudomonas spp. were isolated from wastewater of El‐Malah canal located in Assiut, Egypt and were checked for their heavy metal tolerance. One isolate has tested for its multiple metal resistances and found to be plasmid mediated with molecular weight 27 Kb for nickel and lead. It was identified as Pseudomonas aeruginosa ASU 6a. Its minimal inhibitory concentration (MIC) for Cu2+, Co2+, Ni2+, Zn2+, Cr3+, Cd2+and Pb2+ were 6.3, 5.9, 6.8, 9.2, 5.8, 4.4, and 3.1 mM, respectively. Growth kinetics and the maximum adsorption capacities were determined under Ni2+ and Pb2+ stress. The latter heavy metals induced potassium efflux and were used as indicator for plasma membrane permeabilization. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

Effects of cadmium stress on growth, morphology, and protein expression in Rhodobacter capsulatus B10

The effects of cadmium stress on growth, morphology, and protein expression were investigated in Rhodobacter capsulatus B10 using two-dimensional polyacrylamide gel electrophoresis and a scanning electron microscope with an energy dispersive X-ray spectrometer. The bacterium grew in the presence of 150 μM CdCl2 and highly induced heat-shock proteins (GroEL and Dnak), S-adenosylmethionine synthetase, ribosomal protein S1, aspartate aminotransferase, and phosphoglycerate kinase. Interestingly, the ribosomal protein S1 was proportionally expressed as the amount of cadmium in the medium, suggesting that S1 may be required for the repair of cadmium-mediated cellular damage. On the other hand, we identified five cadmium-binding proteins: 2-methylcitrate dehydratase, phosphate peripalsmic binding protein, inosine-5′-monophosphate dehydrogenase/guanosine-5′-monophosphate reductase, inositol monophosphatase, and lytic murein transglycosylase. The cadmium-treated cells had a filamentous structure and contained less phosphorous than the untreated cells. We propose that these characteristics of the cadmium-treated cells may be due to the inactivation of the phosphate peripalsmic binding protein and lytic murein transglycosylase by cadmium.

Effect of temperature and soil moisture on the survival and symbiotic effectiveness of Frankia spp.

Abstract Comparison of the effects of temperature on the growth in culture (increase in protein) of Frankia showed that three strains isolated from Casuarina were more tolerant of high temperature (45°C) than a strain from Alnus rubra. Optimal temperatures for growth of the Casuarina strains were in the range 25–30°C. Growth of the Alnus strain was good at 25°C but poor at 37°C. High temperatures (35–40°C) during storage for 7 months of these Frankia strains in sand, inoculated initially with liquid culture or with Frankia incorporated into alginate beads and permitted to dry, resulted in substantial loss of infectivity for the host plant species. Loss in infectivity was greater with an Alnus Frankia strain than strains from Casuarina cunninghamiana, C. equisetifolia and C. junghuniana. Three Frankia strains from C. equisetifolia were incorporated into a sand/perlite mixture with three different moisture regimes (field moisture capacity – wet: watered and maintained at field capacity; watered to field capacity but then allowed to dry – moderately wet; or watered to half field capacity and then permitted to dry – dry) and then stored for 12 weeks at 25°C and 35°C. Assessment by the most probable number (MPN) technique of the infectivity of the sand mixture for nodulation of C. equisetifolia showed significant interactions between Frankia strain, temperature and soil moisture content. The infectivity of Frankia strains ORS020607 and UGL020602q was not affected by incubation in wet sand at 25°C but fell by more than half after 12 weeks in moderate and dry conditions. Changes in infectivity were similar when incubation was at 35°C. By contrast, the infectivity of UGL020603q fell substantially under all moisture conditions and at both temperatures. The data show the importance of screening for tolerance of both temperature and moisture content when selecting strains for preparation of inoculum for use in hot climates.

Biodegradation Ability and Catabolic Genes of Petroleum-Degrading Sphingomonas koreensis Strain ASU-06 Isolated from Egyptian Oily Soil

Polycyclic aromatic hydrocarbons (PAHs) are serious pollutants and health hazards. In this study, 15 PAHs-degrading bacteria were isolated from Egyptian oily soil. Among them, one Gram-negative strain (ASU-06) was selected and biodegradation ability and initial catabolic genes of petroleum compounds were investigated. Comparison of 16S rRNA gene sequence of strain ASU-06 to published sequences in GenBank database as well as phylogenetic analysis identified ASU-06 as Sphingomonas koreensis. Strain ASU-06 degraded 100, 99, 98, and 92.7% of 100 mg/L naphthalene, phenanthrene, anthracene, and pyrene within 15 days, respectively. When these PAHs present in a mixed form, the enhancement phenomenon appeared, particularly in the degradation of pyrene, whereas the degradation rate was 98.6% within the period. This is the first report showing the degradation of different PAHs by this species. PCR experiments with specific primers for catabolic genes alkB, alkB1, nahAc, C12O, and C23O suggested that ASU-06 might possess genes for aliphatic and PAHs degradation, while PAH-RHDαGP gene was not detected. Production of biosurfactants and increasing cell-surface hydrophobicity were investigated. GC/MS analysis of intermediate metabolites of studied PAHs concluded that this strain utilized these compounds via two main pathways, and phthalate was the major constant product that appeared in each day of the degradation period.

Antimicrobial activity of latex silver nanoparticles using Calotropis procera

ABSTRACT Objective To synthesize silver nanoparticles (AgNPs) by green methods using serum latex of Calotropis procera at 80 °C and evaluate them against bacteria, dermatophytes and phytopathogenic fungi comparing with the activity of untreated latex. Methods The synthesis of AgNPs was performed by mixing 3% latex serum extract with the same volume of silver nitrate (2 mmol/L) solution in round flask and heating in water bath at 80 °C. Characterization of silver particles were determined using UV-vis spectrophotometer, transmission electron microscopy (TEM), X-ray diffraction and Fourier transform infrared spectroscopy. The antimicrobial activity of the green synthesized AgNPs was determined against bacteria, dermatophytes and phytopathogenic fungi and compared to the crude untreated latex by agar-well diffusion methods. Results Biosynthesis of latex silver nanoparticles was successfully obtained by green method. The formation of AgNPs has been confirmed by UV-vis, TEM microscopy, X-ray diffraction and Fourier transform infrared spectroscopy. TEM analysis showed that synthesized AgNPs are highly stable spherical shaped particles, well dispersed with a diameter ranged from 4 nm up to 25 nm and an average size of 12.33 nm. AgNPs showed strong antibacterial activity against Gram-negative bacteria ( Escherichia coli , Pseudomonas aeruginosa and Serratia sp.) and antifungal activity against Trichophyton rubrum , Candida albicans and Aspergillus terreus . Conclusions It can be concluded that serum latex of Calotropis procera was found to display strong potential for the synthesis of AgNPs as antimicrobial agents through rapid reduction of silver ions (Ag + to Ag 0 ). The green synthesized AgNPs were found to show higher antimicrobial efficacy than crude latex.

Utilization of aromatic compounds by phototrophic purple nonsulfur bacteria

Biodegradation of aromatic compounds byRhodopseudomonas blastica andRhodospirillum rubrum appears to be lacking in the literature. The above species grew phototrophically (illuminated anaerobic conditions) on a variety of organic compounds. They were found to degrade benzoate, benzyl alcohol, 4-hydroxy-3,5-dimethoxybenzoate (Syringate) and 4-hydroxy-3-methoxybenzoate (vanillate). The ability of the above species to photocatabolize aromatic compounds indicates that these organisms may be ecologically significant as scavengers of aromatic derivatives in illuminated anaerobic habitats in nature.

Antioxidant and antiapoptotic activities of Calotropis procera latex on Catfish (Clarias gariepinus) exposed to toxic 4-nonylphenol.

Calotropis procera L. is known as medicinal plant. The Phytochemical analyzes of its latex revealed that it possessed antioxidants, namely terpenes, phenolic compounds and cardenolides, flavonoids and saponins, while tannins, alkaloids and resin were absent in moderate to high concentration. In the present study, the role of latex of Calotropis procera as antioxidant and antiapoptotic was reported. To carry out this aim, fishes were exposed to 100 µg l(-1) 4-nonylphenol as chemical pollutant. The enzymes, superoxidase dismutase, catalase, acetlycholinstrase (AchE), glutathione s-transferase, cortisol, G6PDH) and apoptotic cells increased significantly (p<0.05) accompanied by irregular disturbance of (Na(+), K(+)) ions in the presence of 4-nonylphenol. On the other hand, these enzymes, ions, and apoptotic cells decreased normally and significantly (p<0.05) in the presence of latex. Total phenol content, total capacity antioxidant, reducing power decrease significantly (p<0.05) in the presence of 4-nonylphenol and increase normally in the presence of latex. Latex was used for the first time to protect catfish after 4-nonylphenol exposure. Our study confirms that crude latex of Calotropis procera possessed antioxidant and antiapoptotic activities against the toxicity of 4-Nonylphenol.

Keratinophilic and other fungi isolated from combine harvester wheat and sorghum dusts and from the atmosphere of winnow sites in Egypt.

Three methods were used for the isolation of fungi in the present investigation: the dilution-plate method, hair-baiting technique with horse hairs and the “exposed plate” method. Sabourauds glucose agar at 28 °C was also used as isolation medium. 102 species and 2 species varieties belonging to 36 genera were collected from combine harvester wheat and sorghum dusts and from the atmosphere of hay or winnow sites.Chrysosporium was represented by 6 species:C. asperatum, C. indicum, C. keratinophilum, C. merdarium, C. pannorum andC. tropicum. Several filamentous fungi tolerating high levels of cycloheximide and other moulds were frequently encountered in the two types of dusts and in the two atmospheres, such as members ofAcremonium, Alternaria, Aspergillus, Cladosporium, Emericella, Eupenicillium, Fusarium, Paecilomyces, Penicillium, Piedraia, Rhizopus, Scopulariopsis and others.

Study of enhancement and inhibition phenomena and genes relating to degradation of petroleum polycyclic aromatic hydrocarbons in isolated bacteria

Polycyclic aromatic hydrocarbons (PAHs) are xenobiotic compounds, which being degraded by chemical, physical or biological methods. The latter is the safest and the cheapest one. Two bacterial strains ASU-01 and ASU-016 were isolated from different Egyptian petroleum contaminated sites. They were genetically identified based on the analysis of the nucleotide sequences of the 16S ribosomal PNA gene and the phylogenetic tree as Enterobacter hormaechei and Pseudomonas pseudoalcaligenes respectively. When pyrene as high molecular weight (HMW)-PAH was added as a sole carbon source, both strains could degrade it with efficiency 77.7 and 83.7% within 15 days of incubation, respectively. However, when it was mixed with low molecular weight (LMW)-PAHs, two opposite phenomena appeared. The first one was enhancement, which occurred with ASU-01. This strain shifted pyrene efficiency to 98.5%. The second phenomenon was inhibition occurred with ASU-016 which completely retarded pyrene degradation. Naphthalene dioxygenase (nahAc), and catechol dioxygenases (C12O and C23O) genes were detected in the two strains based on PCR. The detected genes were confirmed by determining the different specific activities of their translated protein (enzymes) on different PAHs. The maximum values of biosurfacatant production activity and cell-surface and percentage of cell-surface hydrophobicity (CSH) were detected during the exponential phase. These latter factors increased the bioavailability and consequently, the assimilation of PAHs.

Cytotoxic cardenolides from the latex of Calotropis procera

Three new cardenolides (3, 9 and 10), along with eight known ones, were isolated from the latex of Calotropis procera. The structural determination was accomplished by the 1D- and 2D-NMR spectra as well as HRESIMS analysis. The growth inhibitory activity of the latex and its sub-fractions as well as isolated compounds was evaluated against human A549 and Hela cell lines. The results exhibited that latex had strong growth inhibitory activity with IC50s of (3.37 μM, A-549) and (6.45 μM, Hela). Among the four extracts (hexane, chloroform, ethyl acetate and aqueous), chloroform extract displayed the highest potential cytotoxic activity, with IC50s of (0.985 μM, A-549) and (1.471 μM, Hela). All the isolated compounds displayed various degrees of cytotoxic activity and the highest activity was observed by calactin (1) with IC50s values of (0.036 μM, A-549) and (0.083 μM, Hela). None of these isolated compounds exhibited good antimicrobial activity evaluated by determination of their MICs using the broth microdilution method against various infectious pathogens. The structure-activity relationships for cytotoxic activity were also discussed.