Fifteen local isolates of Pseudomonas were obtained from several sources such as soil, water and some high-fat foods (Meat, olives, coconuts, etc.). The ability of isolates to produce lipase was measured by the size of clear zone on Tween 20 solid medium and by measuring the enzymatic activity and specific activity. Isolate M3 (as named in this study) was found to be the most efficient for the production of the lipase with enzymatic activity reached 56.6 U/ml and specific activity of 305.94 U/mg. This isolate was identified through genetic analysis of the 16S rRNA gene. and it was shown that the isolate M3 belongs to Pseudomonas aeruginosa with 99% similarity. The DNA of isolate M3 was extracted and lipase gene was amplified through PCR technique, then purified and cloned into E.coli DH5α cells first using pTG19-T plasmid, and expressed in E.coli Bl21 with expression vector pet-28a. The activity of lipase from transformed E.coli Bl21 was 196.6 U/ml and the specific activity 618.2 U/mg.
Background: Pseudomonas aeruginosa is a devious pathogen with the tendency to prompt many acute and serious chronic diseases. This study aims to detect novel genes (Toxins-Antitoxins II system), especially; higB and higA encoded from P. aeruginosa by PCR technique and the relation between these genes and antibiotic resistance of P. aeruginosa. Methods: This study detected 50 isolates of P. aeruginosa from distinct clinical sources. The most common origin of isolates was (44%) burn swabs, (22%) urine culture, (12%) wound swabs, (14%) sputum, and (8%) ear swabs. The bacteria were isolated using implantation MacConkey agar and blood agar, as well as biochemical tests including oxidase test, catalase test then VITEK-2 System of P. aerug
... Show MoreThe bile salt hydrolase gene (bshA), encoding bile salt hydrolase enzyme (EC 3.5.1.24) from probiotic isolate Lactobacillus acidophilus Ar strain which is responsible for assimilation cholesterol were studied in the present work. About 801 bp in length DNA fragment of Lb. acidophilus Ar strain was amplified by PCR techniques. Two restriction sites (PstI/SacI) were added to each end of that fragment for manipulation of DNA during cloning. Amplified fragment inserted into pJET1.2\blunt end vector and pMG36e vector respectively. pJET1.2\blunt end vector is overexpression plasmid for E. coli MC1022, and pMG36e vector is a shuttle vector which is able to replicate in both E. coli and lactic acid bacteria. The resulted constructs were named as pJ
... Show MoreOwing to high antibacterial resistance of Pseudomonas aeruginosa, it could be considered as the main reason behind the nosocomial infections. P. aeruginosa has a well-known biofilm forming ability. The expression of polysaccharide encoding locus (pelA gene) by P. aeruginosa is essential for this ability. The purpose of the current research was to determine the biofilm formation in P. aeruginosa isolated from clinical samples and to evaluate the role of the selected PelA gene in biofilm formation using PCR method in Iraqi patients. Results revealed that 24 (96%) isolates were found to have the ability to form biofilm that was remarkably related to gentamicin resistance. Moreover, the pelA gene was found in all biofilm-producers. In c
... Show MoreThe current study includes 144 samples were 106 bacterial samples belonging to the clinical sources, 38 bacterial samples belonging to the environmental sources to investigate the presence of bacteria P. aeruginosa. The results of diagnosis clarified that there are 45 bacterial isolates belonging to the bacterium P. aeruginosa The examination of the sensitivity of all bacterial isolates was done for elected 45 isolation towards the 11 antibiotic by spread method on the dishes. The results showed that the resistance ratio toward Cefixim, Cefotaxim, Tetracycline, Amoxicillin, Cloxacillin, Methicillin, Erythromycin and Naldixic acid was 77.7, 73.3, 84.4, 82.2, 80, 77.7, 77.7 and 73.3 respectively, While most isolates were sensitive to all o
... Show MoreThis study ,the samples were collected from "118 patients " suffering from burn wound contaminated with Pseudomonas aeruginosa and 100 health individuals (male and female ) as a control group ,the samples were wound swap and blood sample . Chromatography technique was employed to extract and purify cell wall containing lipopolysaccharide by using P. aeruginosa isolate ATCC 15692,the purification done by addition of ammonuium sulfate, sodium dodecyl sulfat (SDS) anddialysis, gel filtration chromatography by using sepharose-4B. Immunogenicity of LPS component was determined by mice injection under the skin ,then Ab concentration agai
... Show MoreOne of the most important virulence factors in Pseudomonas aeruginosa is biofilm formation, as it works as a barrier for entering antibiotics into the bacterial cell. Different environmental and nutritional conditions were used to optimize biofilm formation using microtitre plate assay by P. aeruginosa. The low nutrient level of the medium represented by tryptic soy broth (TSB) was better in biofilm formation than the high nutrient level of the medium with Luria Broth (LB). The optimized condition for biofilm production at room temperature (25 °C) is better than at host temperature (37 °C). Moreover, the staining with 0.1% crystal violet and reading the biofilm with wavelength 360 are considered essential factors in
... Show MoreOver the past few decades, the health benefits are under threat as many commonly used antibiotics have become less and less effective against certain illnesses not only because many of them produce toxic reactions but also due to the emergence of drug-resistant bacteria. The clinical use of a combination of antibiotic therapy for Pseudomonas aeruginosa infections is probably more effective than monotherapy. The present study aims to estimate the antibacterial and antibiofilm activity of Conocarpus erectus leaves extracts against multi-drug resistant P. aeruginosa isolated from different hospitals in Baghdad city. One hundred fifty different clinical specimens were collected from patients from September 2021 to January 2022. All samples were
... Show MoreThis research was conduct to evaluate the cytotoxic effect of exotoxin A (ETA) produced by Pseudomonas aeruginosa on mice in comparison with (phosphate buffer saline (PBS) as a negative control. The effect of the toxin was measured by employing the cytogenetic analysis which included (the mitotic index (MI), chromosomal aberrations (CAs), micronucleus (MN) and sperm abnormalities) parameters. In order to specify the cytotoxic effect of the toxin, three doses of ETA (125, 250 and 500 ng/ml) were used. Results showed that ETA was found to cause a significant decrease in mitotic index (MI) percentage, while significant increase in micronucleus (MN), chromosomal aberrations (CAs) and sperm abnormalities parameters in compression with control wa
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