Burkholderia cepacia and Pseudomonas aeruginosa were isolated from (100) samples collected from patients suffering from burns, wound, sputum, ear swab, curate, operation room, blood, cystic fibrosis, urine and identified using different morphological and biochemical tests as well as using API 20E identification system. • Ability for production of different enzymes was tested and was found that all isolates were alkaline protease, gelatinase and siderphore producer. However, B. cepacia shows higher production than P. aeruginosa. It was found that (78%) and (87.23%) of isolates were able to produce urease and lipase respectively. Complete blood hemolysis (b- hemolysis) occur with most isolates with some variations. • Growth characteristics of both bacterial isolates were studied, and found on log phase the cell number reach about (8×105) cells/ml for B. cepacia, while P. aeruginosa the cell number reach about (4.5×106) cells/ml. • All isolates were subjected to the sensitivity test against thirteen different antibiotic disks. Results show that most of the isolates were resistance to large number of antibiotics.Amikacin and ceftazidime were the most effective antibiotics against the isolates. • Depending on the antibiotic sensitivity results, six isolates were selected for determining the minimal inhibitory concentration (MIC) of some antibiotics. Results showed that bacterial isolates were able to grow at high concentration (1042 mg/ml) of piperacillin for P. aeruginosa and (600 mg/ml) for B. cepacia, and moderate concentration (512, 16mg/ml) of ceftazidime for P. aeruginosa and B. cepacia, with slight resistance for ciprofloxacin (8, 0.5mg/ml). • Inhibitory activity of Lactobacillus acidophilus was determined on solid medium for six bacterial isolates in two incubation periods. It was found that the diameter of zone of growth inhibition is ranged between (10-20mm) after 24h of incubation. While in the liquid media it was found that L. acidophilus filtrate (non-concentrated) have no effect on both isolates, therefore four folds concentrated filtrates of LAB was used and tested at three incubation periods. It was found that three-fold and four- fold concentrated filtrates exhibit the highest inhibitory effects at 24 and 48h of incubation as the diameter of the three-fold between (18-20mm) and at the four-fold it reach to (23mm) inhibitory zone. • MIC of LAB concentrated filtrate was estimated, results showed that (50%) concentration was effective to inhibit bacterial growth. MICs were determined for the fourfold concentrated filtrate of LAB against adhesion property of isolates (R17 and R1). Results showed that filtrates were able to lower adhesion to the epithelial cells as indicated by lowerness in average of adherence from (55-70) bacteria/cell before treatment to (10-14) bacteria/cell after treatment.• Productivity of selected isolates for alginate was determined; such that an amount of 106mg/ml and 115mg /ml of alginate were estimated for isolate R44 and R17 which belong to B. cepacia. • Genetic analysis of adhesive factors in both bacterial isolates was done using isolation of plasmid and chromosomal DNA, plasmid curing and transformation. • Plasmid profile for B. cepacia (R17, R18, R23, R28, R44) and P. aeruginosa (R1, R2, R3, R7, R14, R16, R27) were done; it was found that only two small plasmid bands were characterized on the gel for all isolates. • Isolation of chromosomal DNA from B. cepacia (R17, R23, R28, R44) and P. aeruginosa (R7, R14, R16) isolates was successful and clean chromosomal bands on the agarose gelwere characterized for all isolates. • Curing of plasmids was made, using two curing agents ethidium bromide and SDS; all concentrations were found uneffective in curing B. cepacia plasmid DNA. However (1200 mg/ml) and (0.05g/ml) of ethidium bromide and SDS were found to been effective concentrations for carrying out the curing on P. aeruginosa isolates. • Transformation of plasmid DNA and chromosomal DNA isolated from B. cepacia and P. aeruginosa used as donors to negative strain E. coli were done. No transformants were obtained carrying plasmid DNA. However, number of transformants were obtained using chromosomal DNA with a frequencies of (5.6×103) and (2.1×102) for P. aeruginosa and B. cepacia respectively. • The purity of DNA was determined for B. cepacia (R17, R23, R28, R44) and P. aeruginosa (R7, R14, R16) isolates; the range was (1.6982 and 1.8787mg/ml) for R7 and R16 respectively, DNA concentration was also determined the lowest value was found (5.6707mg/ml) for isolate R7 and the highest value was (14.885mg/ml) for R16 while protein concentration was found (32.818mg/ml) as the highest reading for isolate R28 and (21.299mg/ml) as the lowest value for isolate R7. • Molecular analysis was also performed using Random Amplified Polymorphic DNAPolymerase Chain Reaction (RAPD-PCR) assay; ten different primers chosen randomly were used to analyze bacterial DNA. The first four primers (OPC-5, OPD-18, OPE-13, and OPF-19) showed no amplification results; while the other six primers (OPA-18, OPF-6, OPF-13, OPF-16, OPN-16, and OPO-16) varies in amplification and production of bands. OPA-18 primer shows ten bands with molecular weights ranged (0.4-1.65Kbp) for isolates (R16, R17 and R23). While OPF-6 primer, 18 bands were amplified with molecular weights ranged (0.05-2.45Kbp), was detected among B. cepacia isolates (R44, R28, and R23). Using OPF-13 primer 47, bands were amplified with molecular weights ranged between (0.1- 15Kbp). While using OPF-16 primer only four bands were developed with molecular weights (5, 8, 10, 15) Kbp for isolate R28; and using OPN-16 primer only one band (0.4 Kbp) was detected for R14 (P. aeruginosa) isolates. OPO-16 primer gave18 bands with molecular weights ranged (0.4-2Kbp) for (R44, R28, R23, R17, and R16) isolates. • Efficiency and discriminatory power for primers were calculated and it was found that highest primer efficiency and discriminatory power value were found as (47.9%) and (54.5%) for OPF-13 primer respectively. • Our strategy was to elute specific bands from the gel of (RAPD-PCR) products do be transformed in E. coli HB101. As long as no monomorphic bands were obtained with these primers, no transformation experiment was done.