One hundred isolates of Pseudomonas aeruginosa were obtained from patients admitted to Baghdad hospitals, Iraq during the period between May 2018 until July 2018. These isolates were distributed as 15 isolates from blood, 25 isolates from urinary tract infections, 10 isolates from sputum, 12 isolates from wounds, 15 isolates from ear infections, 15 isolates from bronchial wash of patients suffering from respiratory tract infections in addition to 8 isolates from cystic fibrosis patients. The isolates were initially identified by culturing on MacConkey agar, blood agar and P. aeruginosa agar then diagnosed by performing some morphological and biochemical tests. The second diagnosis was done by API 20E system followed by Vitek 2 compact system. Antibiotic sensitivity test was carried out towards 15 antibiotics using disc diffusion method (Kirby–Bauer method). The results of sensitivity test showed that P. aeruginosa isolates possessed high resistance towards most antibiotics under study, the most antibiotic resistance was towards Gentamicin 87 (87%), whereas the lowest resistance was towards Imipenem 10 (10%). In this study, two types of methods were used in the detection of biofilm formation: the first one was Congo red agar method and the second one was microtiter plate method. In the first method, results showed that biofilm formed by 57/100 (57%) according to black color production on media, whereas in the second method was 69/100 (69%) produce strong adherence according to OD in ELISA reader. Genotypic detection of many virulence factors related to P. aeruginosa was performed using conventional PCR. These included: gene coded for exoenzyme S (exoS), exoenzyme U (exoU), exotoxin A (toxA), two phospholipases C encoded by (plcH) and (plcN), alginate (algD), (lasB), rpsl, proteaseIV, and Neuraminidase (nan1). The results revealed that the most frequent gene was exoS as it was detected in 87/100 (87%) isolates, whereas the least frequent gene was nan1 as it was detected in only 9/100 (9%). The frequency of detection of other genes were as follows: toxAi in 55/100 (55%); plcH in 45/100 (45%); exoU in 42/100 (42%); plcN in 33/100 (33%); proteaseIV in 31/100 (31%), algD in 29/100 (29%); lasB in 28/100 (28%), and rpsl in 25/100 (25%). Phylogenetic analysis by Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR), ERIC-DNA Fingerprinting revealed the diversity of all isolates in Baghdad by using Dice coefficient and the unweighted pair group method with arthmetic average (group method) of phylogenetic analysis. The percentage level of similarity clearly showed that the isolates examined by species were divided into two distinct cluster numbers, in addition to three single isolates (clone), that clustered at a similarity level of (93%). According to the statistical analysis, it was found that the correlation coefficient of ERIC genotyping method with virulence genes in this study and antibiotics sensitivity test was significant at P < 0.05 (two-tailed), whereas correlation with biofilm was not significant
Pseudomonas aeruginosa is an opportunistic pathogen responsible for serious infections. At least three different exopolysaccharides, alginate, polysaccharide synthesis locus (Psl), and pellicle exopolysaccharide (Pel) make up the biofilm matrix in P. aeruginosa . The effect of temperature on the biofilm formation and gene expression was examined by microtiter plate and real-time quantitative polymerase chain reaction (qRT-PCR). To be able to determine the effect of temperature on biofilm formation and gene expression of P. aeruginosa, 303 clinical and environmental samples were collected. Pseudomonas aeruginosa was isolated from 61 (20.1%) and 48 (15.8%) of the clinical and e
... Show MoreFifteen 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 tec
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In this study, 20
This study is concerned with a survey of seven species belonging to seven genera under two families and two orders found in some different areas of the Tigris River, especially since these areas have not been surveyed for a long time, and an attempt to identify the existing species at the present time after the recent water scarcity of the Tigris and Euphrates rivers and all water bodies interior of Iraq and the impact of this scarcity on the fish diversity found in some areas of the Tigris River in Baghdad.
Peroxidase is a class of oxidation-reduction reaction enzyme that is useful for accelerating many oxidative reactions that protect cells from the harmful effects of free radicals. Peroxidase is found in many common sources like plants, animals and microbes and have extensive uses in numerous industries such as industrial, medical and food processing. In this study, P. aeruginosa was harvested to utilize and study its peroxidases. P. aeruginosa was isolated from a burn patient, and the isolate was verified as P. aeruginosa using staining techniques, biochemical assay, morphological, and a sensitivity test. The gram stain and biochemical test result show rod pink gram-ne
... Show MoreThis work aimed to use conventional PCR to identify Salmonella spp. that were isolated from diarrheal children and healthy and diarrheic dogs based on four virulence genes, hilA, stn, spvR, and marT. Sixteen Salmonella isolates including: 9 isolated from children's diarrhea from three species (S. Typhimurium, S. Enteritidis, S. Typhi) and seven isolated from dogs including (S. Typhimurium, S. Enteritidis, S. Muenchen), were identified primarily by several methods. The PCR products of the 16S rRNA gene were sequenced and examined using BLAST analysis to find differences and similarities between these Iraqi isolates and already-known global strains in order to construct the phylogenetic tree of S.
... Show MoreAntibiotics present the greatest threat to soil and aquatic ecosystems among the different therapeutic groups of medicines (which include prescription drugs and treatments for cancer). The strongest drugs, antibiotics, have been utilized to stop the growth of microorganisms or eradicate them. Using high-performance liquid chromatography technology with fluorescence detection, the amounts of levofloxacin and tetracycline in the wastewater from three hospitals (Medical City, Al-Kindi, and Al-Yarmouk) were determined. Levofloxacin and tetracycline were chosen in this study because they are the most important water pollutants. These antibiotic residues were separated and measured using a gradient elution technique on a reverse-phase C18
... Show MoreAntibiotics present the greatest threat to soil and aquatic ecosystems among the different therapeutic groups of medicines (which include prescription drugs and treatments for cancer). The strongest drugs, antibiotics, have been utilized to stop the growth of microorganisms or eradicate them. Using high-performance liquid chromatography technology with fluorescence detection, the amounts of levofloxacin and tetracycline in the wastewater from three hospitals (Medical City, AlKindi, and Al-Yarmouk) were determined. Levofloxacin and tetracycline were chosen in this study because they are the most important water pollutants. These antibiotic residues were separated and measured using a gradient elution technique on a reverse-phase C18 co
... Show MoreP. aeruginosa is a famous bacterium that causes several diseases and has a high ability to be a multidrug resistant organism that is linked with the formation of biofilm. This study aimed to investigate tssC1 gene role in the resistance of different antibiotics in the presence of biofilm. We constructed biofilm for the isolates under the study and showed the effect of different antibiotics on biofilm formation and maturation. The presence of the gene was detected through achieving PCR reaction. Finally, tssC1 gene variation was determined through sequencing and aligning the sequencing products. The results showed that most of the isolates (80%) formed biofilm that played a role in the resistance of different antibiotics which could
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