Antioxydant and Antibacterial Activity of Alkaloids and Terpenes Extracts from Euphorbia granulata

In order to enhance the knowledge of certain phytochemical Algerian plants that are widely used in traditional medicine and to exploit their therapeutic potential in modern medicine, we have done a specific extraction of terpenes and alkaloids from the leaves of Euphorbia granulata to evaluate the antioxidant and antibacterial activity of this extracts. After the extraction it was found that the terpene extract gave the highest yield 59.72% compared with alkaloids extracts. The disc diffusion method was used to determine the antibacterial activity against different bacterial strains: Escherichia coli (ATCC25922), Pseudomonas aeruginosa (ATCC27853) and Staphylococcus aureus (ATCC25923). All extracts have shown inhibition of growth bacteria. The different zones of inhibition have varied from (7 -10 mm) according to the concentrations of extract used. Testing the antiradical activity on DPPH-TLC plates indicated the presence of substances that have potent anti-free radical. As against, the BC-TLC revealed that only terpenes extract which was reacted positively. These results can validate the importance of Euphorbia granulata in traditional medicine.

Therapeutic Product Preparation Bioprocess Modeling

An immunomodulator bioproduct is prepared in a batch bioprocess with a modified bacterium Pseudomonas aeruginosa. The bioprocess is performed in 100 L Bioengineering bioreactor with 42 L cultivation medium made of peptone, meat extract and sodium chloride. The optimal bioprocess parameters were determined: temperature – 37 0C, agitation speed - 300 rpm, aeration rate – 40 L/min, pressure – 0.5 bar, Dow Corning Antifoam M-max. 4 % of the medium volume, duration - 6 hours. This kind of bioprocesses are appreciated as difficult to control because their dynamic behavior is highly nonlinear and time varying. The aim of the paper is to present (by comparison) different models based on experimental data. The analysis criteria were modeling error and convergence rate. The estimated values and the modeling analysis were done by using the Table Curve 2D. The preliminary conclusions indicate Andrews-s model with a maximum specific growth rate of the bacterium in the range of 0.8 h-1.

Prevention of Biofilm Formation in Urinary Catheter by Coating Enzymes/ Gentamycin/ EDTA

Urinary Tract Infections (UTI) account for an estimated 25-40% nosocomial infection, out of which 90% are associated with urinary catheter, called Catheter associated urinary tract infection (CAUTI). The microbial populations within CAUTI frequently develop as biofilms. In the present study, microbial contamination of indwelling urinary catheters was investigated. Biofilm forming ability of the isolates was determined by tissue culture plate method. Prevention of biofilm formation in the urinary catheter by Pseudomonas aeruginosa was also determined by coating the catheter with some enzymes, gentamycin and EDTA. It was found that 64% of the urinary catheters get contaminated during the course of catheterization. Of the total 6 isolates, biofilm formation was seen in 100% Pseudomonas aeruginosa and E. coli, 90% in Enterococci, 80% in Klebsiella and 66% in S. aureus. It was noted that the biofilm production by Pseudomonas was prolonged by 7 days in amylase, 8 days in protease, 6 days in lysozyme, 7days in gentamycin and 5 days in EDTA treated catheter.

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.

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.