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
Rice is a major staple food for more than two thirds of the world population. Pathogenesis-related proteins-10 (PR10) have a range of 154 to 163 amino acid with molecular weight ~ 17 kDa. They are acidic and generally intracellular and cytosolic proteins accumulate in plants in response to biotic and abiotic stresses. In the present study, a PR10 gene and its corresponding protein were characterized in O. sativa, O. barthii, O. glaberrima, O. glumipatula, O. meridionalis, O. nivara, O. rufipogon and O. punctata. The results revealed a narrow range of variation at both DNA and protein levels in all examined species except O. glumipatula. The latter showed a relatively
... Show MoreUropathogenic Escherichia coli is the main cause of urinary tract infections, the ability of this bacteria to cause urinary tract infections is related to a variety of virulence factors that enhance colonization and evade the immune response, one of these virulence factors is cytotoxic necrotizing factor 1 toxin which converts the glutamine residue to glutamic acid to activated GTPase Rho family. The study was meant to find out the prevalence rate of the cnf1 gene in Uropathogenic Escherichia coli isolated from Iraqi patients. Conventional laboratory methods were used for primary bacterial identification and molecular methods were used to confirm bacterial identity and gene detection. Escherichia coli was identified in 89/165 (53.93%) of th
... Show MoreThis study aimed at isolating uropathogenic Escherichia coli from urinary tract infections (UTIs) of human and cattle to examine the molecular diversity and phylogenetic relationship of the isolates. A total of 100 urine samples were collected from UTIs of human and cattle. The isolates identification was done using routine diagnostic methods and confirmed by Vitek2. Antimicrobial susceptibility was tested against 10 antimicrobials. Random amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) was applied to identify the genetic diversity among E. coli isolates from human and animal origin by using five different octamer primers. The gelJ software for the phylogenetic analysis created Dendrograms. Out of 50 human urine samples, E.
... Show MoreTen isolates of Klebsiella pneumoniae, seven isolates of Pseudomonas aeruginosa and nine isolates of Staphylococcus aureus, were obtained from 100 urine samples collected from Baghdad hospitals. All isolates were identified biochemically and confirmed by using VITEK 2 and were then tested for their susceptibility towards 6 antibiotics and for phenolic extracts of Thymus vulgaris and Cinnamomum cassia. All bacteria were greatly affected by T. vulgaris, especially K. pneumoniae. Viable count was performed, it was noted that the number of bacterial cells reduced from 1×108 CFU to 1.2× 103, 2×105 and 1.8×106CFU of K. pneumoniae, P. aeruginosa and S. aureus respectively. While C. cassiahad a slight effect on them. K. pneumoniae isola
... Show MoreBackground: The microbial production of substances that have the potency to suppress the growth of other microorganisms is probably one of the prevalent defense strategy developed in nature, microorganisms produce a variable bunch of microbial defense systems, which include antibiotics, metabolic by-products, lytic agents, bacteriocins and others. Objective: The purpose of the present study was to isolate and identify Enterococcus faecium isolates then detecting its ability of carrying the gene responsible for enterocin production in this species. Materials and methods: Out of 50 samples from different sources (food and clinical sources) were collected for the Enterococcus faecium isolation, and the isolated bacteria Enterococ
... Show MoreSwarming is one of the most important virulence factors used by bacteria to invade new sites. This study aimed to test the effects of gentamicin on swarming motility of Pseudomonas aeruginosa, both phenotypically and molecularly. The present results revealed that 11/25 isolates had gentamicin MIC of 1024 µg/ml. However, gentamicin at sub-minimal inhibitory concentration significantly (P< 0.05) reduced the diameter of swarming in all P. aeruginosa isolates. Noticeably the mean and median swarming diameter before treatment with gentamicin 5.557 and 5.816 cm respectively had significantly (P < 0.001) reduced to 0.871 and 0.766 cm respectively. At the molecular level, amrZ (a global regulator of multiple genes) and
... Show MoreThe optimum cultural conditions for garamicidin production by local isolate B.brevis were studied.Best result was obtained when the isolate B.brevis was grown on media composed of 1%glucose as carbon source,1% ammonium chloride as a nitrogen source ,0.5% Dipotassium hydrogen orthophosphate as a phosphate source and after 48 hours of incubation at 30C .Garamicidin has been extracted and purified through acid precipition and then extracted by organic solvent (ether& acetone ).Using HPLC the garamicidin antibiotic showed three types A,B and C garamicidin .
Reishi Mushroom, Ganoderma, is considered one of important wood-decaying medicinal mushrooms. This study aimed to identify three samples of this genus in Mosul city in February and April 2019. Three species of Ganoderma were collected from three various trees including Eucalyptus, Morus, and Olea (olive) in Mosul City, Northern Iraq. Their identifications and their DNA sequences were genetically identified by using PCR techniques according to detect nuclear ribosomal internal transcribed spacer (ITS) regions. Results exhibited the finding of Ganoderma resinaceum, Ganoderma applanatum, and Ganoderma sp. This study is first attempt to identify Reishi Mushroom by molecular methods in Iraq. Thus, the current study is considered new good d
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