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
Respiratory tract infections in sheep are among the important health problems that affect all sheep ages around the world. Nine bacterial isolates obtained from sheep with respiratory tract infections were selected to be used in the current study. The isolates included 3 Staphylococcus aureus, 4 Klebsiella pneumoniae, and 2 Pseudomonas aeruginosa. Following the primers design by the Primer3Plus software tool and optimization of the conventional polymerase chain reaction (PCR), the primers were validated for their use in the multiplex PCR experiments. The MFEprimer program was used to check the suitability of the primer set combinations for multiplex PCR. The MFEprimer software was successful in designing the multiplex-PCR experiments and de
... Show MoreThe number of infections caused by microorganisms is increasing significantly over the last few years. A total of 140 patients admitted to the central teaching hospital of pediatrics from the 1st of Jun 2017 to 31 October 2017. The Clinical samples was processed from culture and sensitivity testing. Antibiotic discs used for gram negative isolates. The most prevalent gram negative isolates included Escherichia coli 63 (45.0 %), Pseudomonas spp. 21 (15.0 %), Klebsiella spp. 19 (13.6 %) predominantly. Escherichia coli were the most prevalent isolates from urine 45 (71.4 %), Klebsiella spp. 11 (57.9 %) and Enterobacter spp. 11 (68.8 %) followed by Escherichia coli 10 (15.9 %) predominant from blood. 68 (48.6 %) of specimens were urine, 47 (33.
... Show MoreDiabetic mellitus is one of the main risk factors of fungal infections because poor glycemic control is associated with a high level of glucose in blood and saliva which could be treated as nutrient to fungi. This study aimed to isolate and identification of pathogenic fungi from diabetic patient. 140 samples were taken from different places of human body from the national center of diabetic patients that related to Mustansiriyah University / college of medicine and Al-yarmuk Hospital in Baghdad. 84 sample (60%) tested positive to fungi and 56 sample (40%) tested negative to fungi. The most frequented fungi isolated have been chosen for molecular identification by PCR (Millerozyma farinosa and Candida orthopsilosis) using specific pri
... Show MoreThe current study was designed to explore the association between the pigments production and biofilm construction in local Pseudomonas aeruginosa isolates. Out of 143 patients suffering from burns, urinary tract infections (UTI), respiratory tract infections and cystic fibrosis obtained from previous study by Mahmood (2015), twenty two isolates (15.38%) were identified from (11) hospitals in Iraq, splitted into three provinces, Baghdad, Al-Anbar and Karbala for the duration of June 2017 to April 2018. Characterization was carried out by using microscopical, morphological and biochemical methods which showed that all these isolates belong to P. aeruginosa. Screening of biofilm production isolates was carried out by usi
... 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 MoreThe present study was carried to evaluate antibacterial activity of Acetone, Alcholic (cold and hot) and Aqueaus(water) extracts of Citrus aurantifoliaseeds,against growth of some bacteria isolated from burns infections(Pseudomonas aeruginosa,Escherichia coli, Klebsiellapneumonia,Staphylococcusaureus)fromKindy HospitalIn Baghdad from March to June 2012.Antibiotic Sensitivity was done for all isolated bacteria used in this study.Results showed variation in antibacterial activity of different extracts against all tested bacteria by well diffusion technique in agar and measuring the diameter of inhibition zone, at concentration 250Mg-ml. Acetone extract had the greatest inhibitory effect followed by hot alcoholci extract, and then cold alcohol
... Show MoreThe effect of local Lactobacillus gasseri filtrate against Pseudomonas aeruginosa infection in mice was studied . 0.25 ml of concentrated filtrate Lactobacillus gasseri was injected in intraperitoneally ( I.P.) 5 days before challenge with 0.2 ml viable P. aeruginosa ( 10 8 cell/ ml). Animals were sacrificed after 12 h. from challenge by cutting the femoral artery . To follow bacterial growth in the peritoneal cavity , its contents were washed out with 5 ml of PBS .The fluid was diluted, 0.1 ml from each dilution and was spread on culture media. The number of colonies in 5 ml of harvested fluid was expressed as Log 10 CFU ,and the percentage of Macrophage in t
... Show MoreMRSA is one of the major pathogens in hospitals and the community, which have the ability to produce biofilm as a virulence factor, the impact of chalcone on biofilm formation, the synergism effect of chalcone and antibiotic in both in vitro and in vivo experiments, the gene expression of virulence genes (srtA, fnbA, fnbB) before and after treatment of it on MRSA biofilm cells in vitro, all these were the prime aims of this study. Chalcone at MBIC (20 μg/ml), significantly reduced the biofilm formation to 21.45% and at sub MBIC (15 μg/ml) to 36.58 %. While, Chalcone at MIC(5 μg/ml) reduced MRSA planktonic cells to 49.61%. Susceptibility of MRSA isolates against eight antibiotics showed that all isolates were sensitive to vancomycin and n
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