Biological Effects of a Carbohydrate-Binding Protein from an Annelid, Perinereis nuntia Against Human and Phytopathogenic Microorganisms

Lectins have a good scope in current clinical microbiology research. In the present study evaluated the antimicrobial activities of a D-galactose binding lectin (PnL) was purified from the annelid, Perinereis nuntia (polychaeta) by affinity chromatography. The molecular mass of the lectin was determined to be 32 kDa as a single polypeptide by SDS-PAGE under both reducing and non-reducing conditions. The hemagglutinating activity of the PnL showed against trypsinized and glutaraldehyde-fixed human erythrocytes was specifically inhibited by D-Gal, GalNAc, Galβ1-4Glc and Galα1-6Glc. PnL was evaluated for in vitro antibacterial screening studies against 11 gram-positive and gram-negative microorganisms. From the screening results, it was revealed that PnL exhibited significant antibacterial activity against gram-positive bacteria. Bacillus megaterium showed the highest growth inhibition by the lectin (250 μg/disc). However, PnL did not inhibit the growth of gram-negative bacteria such as Vibrio cholerae and Pseudomonas sp. PnL was also examined for in vitro antifungal activity against six fungal phytopathogens. PnL (100 μg/mL) inhibited the mycelial growth of Alternaria alternata (24.4%). These results indicate that future findings of lectin applications obtained from annelids may be of importance to life sciences.

Antibacterial Effect of Silver Nanoparticles on Multi Drug Resistant Pseudomonas Aeruginosa

Multidrug resistant organisms have been taunting the medical world for the last few decades. Even with new antibiotics developed, resistant strains have emerged soon after. With the advancement of nanotechnology, we investigated colloidal silver nanoparticles for its antimicrobial activity against Pseudomonas aeruginosa. This organism is a multidrug resistant which contributes to the high morbidity and mortality in immunocompromised patients. Five multidrug resistant strains were used in this study. The antimicrobial effect was studied using the disc diffusion and broth dilution techniques. An inhibition zone of 11 mm was observed with 10 μg dose of the nanoparticles. The nanoparticles exhibited MIC of 50 μg/ml when added at the lag phase and the subinhibitory concentration was measured as 100 μg/ml. The MIC50 value showed to be 15 μg/ml. This study suggests that silver nanoparticles can be further developed as an antimicrobial agent, hence decreasing the burden of the multidrug resistance phenomena.

Improvement of Lipase Catalytic Properties by Immobilization in Hybrid Matrices

Lipases are enzymes particularly amenable for immobilization by entrapment methods, as they can work equally well in aqueous or non-conventional media and long-time stability of enzyme activity and enantioselectivity is needed to elaborate more efficient bioprocesses. The improvement of Pseudomonas fluorescens (Amano AK) lipase characteristics was investigated by optimizing the immobilization procedure in hybrid organic-inorganic matrices using ionic liquids as additives. Ionic liquids containing a more hydrophobic alkyl group in the cationic moiety are beneficial for the activity of immobilized lipase. Silanes with alkyl- or aryl nonhydrolizable groups used as precursors in combination with tetramethoxysilane could generate composites with higher enantioselectivity compared to the native enzyme in acylation reactions of secondary alcohols. The optimal effect on both activity and enantioselectivity was achieved for the composite made from octyltrimethoxysilane and tetramethoxysilane at 1:1 molar ratio (60% increase of total activity following immobilization and enantiomeric ratio of 30). Ionic liquids also demonstrated valuable properties as reaction media for the studied reactions, comparable with the usual organic solvent, hexane.

Effect of Organic Matter and Biofertilizers on Chickpea Quality and Biological Nitrogen Fixation

In order to evaluation the effects of soil organic matter and biofertilizer on chickpea quality and biological nitrogen fixation, field experiments were carried out in 2007 and 2008 growing seasons. In this research the effects of different strategies for soil fertilization were investigated on grain yield and yield component, minerals, organic compounds and cooking time of chickpea. Experimental units were arranged in split-split plots based on randomized complete blocks with three replications. Main plots consisted of (G1): establishing a mixed vegetation of Vicia panunica and Hordeum vulgare and (G2): control, as green manure levels. Also, five strategies for obtaining the base fertilizer requirement including (N1): 20 t.ha-1 farmyard manure; (N2): 10 t.ha-1 compost; (N3): 75 kg.ha-1 triple super phosphate; (N4): 10 t.ha-1 farmyard manure + 5 t.ha-1 compost and (N5): 10 t.ha-1 farmyard manure + 5 t.ha-1 compost + 50 kg.ha-1 triple super phosphate were considered in sub plots. Furthermoree four levels of biofertilizers consisted of (B1): Bacillus lentus + Pseudomonas putida; (B2): Trichoderma harzianum; (B3): Bacillus lentus + Pseudomonas putida + Trichoderma harzianum; and (B4): control (without biofertilizers) were arranged in sub-sub plots. Results showed that integrating biofertilizers (B3) and green manure (G1) produced the highest grain yield. The highest amounts of yield were obtained in G1×N5 interaction. Comparison of all 2-way and 3-way interactions showed that G1N5B3 was determined as the superior treatment. Significant increasing of N, P2O5, K2O, Fe and Mg content in leaves and grains emphasized on superiority of mentioned treatment because each one of these nutrients has an approved role in chlorophyll synthesis and photosynthesis abilities of the crops. The combined application of compost, farmyard manure and chemical phosphorus (N5) in addition to having the highest yield, had the best grain quality due to high protein, starch and total sugar contents, low crude fiber and reduced cooking time.

Microbiological and Physicochemical Studies of Wetland Soils in Eket, Nigeria

The microbiological and physicochemical characteristics of wetland soils in Eket Local Government Area were studied between May 2001 and June 2003. Total heterotrophic bacterial counts (THBC), total fungal counts (TFC), and total actinomycetes counts (TAC) were determined from soil samples taken from four locations at two depths in the wet and dry seasons. Microbial isolates were characterized and identified. Particle size and chemical parameters were also determined using standard methods. THBC ranged from 5.2 (+0.17) x106 to 1.7 (+0.18) x107 cfu/g and from 2.4 (+0.02) x106 to 1.4 (+0.04) x107cfu/g in the wet and dry seasons, respectively. TFC ranged from 1.8 (+0.03) x106 to 6.6 (+ 0.18) x106 cfu/g and from 1.0 (+0.04) x106 to 4.2 (+ 0.01) x106 cfu/g in the wet and dry seasons, respectively .TAC ranged from 1.2 (+0.53) x106 to 6.0 (+0.05) x106 cfu/g and from 0.6 (+0.01) x106 to 3.2 (+ 0.12) x106 cfu/g in the wet and dry season, respectively. Acinetobacter, Alcaligenes, Arthrobacter, Bacillus, Beijerinckja, Enterobacter, Micrococcus, Flavobacterium, Serratia, Enterococcus, and Pseudomonas species were predominant bacteria while Aspergillus, Fusarium, Mucor, Penicillium, and Rhizopus were the dominant fungal genera isolated. Streptomyces and Norcadia were the actinomycetes genera isolated. The particle size analysis showed high sand fraction but low silt and clay. The pH and % organic matter were generally acidic and low, respectively at all locations. Calcium dominated the exchangeable bases with low electrical conductivity and micronutrients. These results provide the baseline data of Eket wetland soils for its management for sustainable agriculture.

Molecular Identification of ESBL Genesbla GES-1, blaVEB-1, blaCTX-M blaOXA-1, blaOXA-4,blaOXA-10 and blaPER-1 in Pseudomonas aeruginosa Strains Isolated from Burn Patientsby PCR, RFLP and Sequencing Techniques

Fourty one strains of ESBL producing P.aeruginosa which were previously isolated from burn patients in Kerman University general hospital, Iran were subjected to PCR, RFLP and sequencing in order to determine the type of extended spectrum β- lactamases (ESBL), the restriction digestion pattern and possibility of mutation among detected genes. DNA extraction was carried out by phenol chloroform method. PCR for detection of bla genes was performed using specific primer for each gene. Restriction Fragment Length Polymorphism (RFLP) for ESBL genes was carried out using EcoRI, NheI, PVUII, EcoRV, DdeI, and PstI restriction enzymes. The PCR products were subjected to direct sequencing of both the strands for identification of the ESBL genes.The blaCTX-M, blaVEB-1, blaPER-1, blaGES-1, blaOXA-1, blaOXA-4 and blaOXA-10 genes were detected in the (n=1) 2.43%, (n=41)100%, (n=28) 68.3%, (n=10) 24.4%, (n=29) 70.7%, (n=7)17.1% and (n=38) 92.7% of the ESBL producing isolates respectively. The RFLP analysis showed that each ESBL gene has identical pattern of digestion among the isolated strains. Sequencing of the ESBL genes confirmed the genuinety of PCR products and revealed no mutation in the restriction sites of the above genes. From results of the present investigation it can be concluded that blaVEB-1 and blaCTX-M were the most and the least frequently isolated ESBL genes among the P.aeruginosa strains isolated from burn patients. The RFLP and sequencing analysis revealed that same clone of the bla genes were indeed existed among the antibiotic resistant strains.