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
Dual-species biofilms of Pseudomonas aeruginosa and Staphylococcus aureus generate difficult-to-treat illnesses. Nutrition stress in biofilms affects physiology, microbial metabolism, and species interactions, impacting bacteria growth and survival. Furthermore, the function of alginate, which is encoded by the algD gene, in the production of biofilms has been established. The present study aimed at investigating the impact of starvation on algD gene expression in single-species biofilm of P. aeruginosa and dual-species biofilms of P. aeruginosa and S. aureus from hospital sewage. A total of six P. aeruginosa and six S. aureus isolates were obtained from the microbiology laboratory at the Department of Biology, College of Science, Universit
... Show MoreThe following study was conducted to investigate the correlation between the expression of three different genes (NOB1, DDX47, CD101( with the occurrence and development of chronic myeloid leukemia (CML) in Iraq. The difference in the expression of these genes between patients and healthy controls was studied. Moreover the correlation of age and gender with CML occurrence and comparing with control was also examined. Results showed significant increases in mean of gene expression level (ΔCt) of patient groups for all genes compared to the corresponding ΔCt means in control group, also the gene expression folding (2-ΔΔCt) reflect significant differences in the expression of these genes and CD101, mRNA showed the highest level in CML pati
... Show MoreThe increasing use of antiseptic compounds creates selective pressure cause emergence of antiseptic resistance among Staphylococcus aureus .Resistance mechanism of antiseptic is driven mainly by multi drug resistant (MDR) efflux protein.Sixty five isolates of S.aureuswere collected from different clinical sources and subjected to 11 antibiotics most of them are recognized by efflux systems as extruded substrates. Range of efflux activity was estimated using cartwheel method. Simultaneous discrimination of antiseptic coding genes (qacA/B, smr and norA)as well as nuc and mecA genes among multidrug resistantS.aureus(MRSA) isolates was preformed using multiplex PCR assay
... Show MoreObjective Using two complementary techniques of virus detection human papillomavirus (HPV)[capture of hybrids (CH) and polymerase chain reaction (PCR)], relate the cytological study and/or cervical biopsy with high-risk HPV (HPV-HR) genotypes presence, as well as relating their viral load (VL). Methods About 272 women, who presented most cell alterations compatible with lesions cervical HPV, which has been detected in all high risk by the CH method and HPV genotype detection by PCR. Results In 22% of the patients it was not detected HPV DNA. Genotype 16 and/or 18 was prevalent and was found in 33% of the 212 women studied, meanwhile, mixed infections were found by several genotypes in 25%. In as for the histological lesions found, in 61 pat
... Show MoreCandida Berkh. (1923) occurs naturally in the body. But it becomes opportunistic fungi, meaning that it infects humans when there is any weakening of the immune system, such as exposure to chemotherapy, diabetes, or organ transplantation. Most species of Candida grow at a temperature between 20-40 °C and have a pH of 3-8. Human pathogens of Candida species include C. albicans, C. glabrata, C. lusitaniae, C. parapsilosis, C. tropicalis and C. utili. C. albicans has many virulence factors that facilitate injury process. Virulence factors are considered as a measure of pathogenicity, and it is in the form of fungal toxins, enzymes, or cell structures that facilitate infection, as well as pathogen resistance in different conditions. This study
... Show MoreUrinary tract infection is a bacterial infection that often affects the bladder and thus the urinary system. E. coli is one of the leading uropathogenic bacteria that cause urinary tract infections. Uropathogenic E. coli is highly effective and successful in causing urinary tract infections through biofilm formation and urothelial cell invasion mechanisms. Other organisms that cause urinary tract infections include members of the Enterobacteriaceae family, streptococci and staphylococci species and perch. In addition, K.penumoniae is another important gram-negative bacterium that causes urinary tract infections. With the PCR technique, unseen bacterial species can be detected using standard clinical microbiology methods. In this study, the
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Detection of virulence gene agglutinin-like sequence (ALS) 1 by using molecular technology from clinical samples (
Pseudomonas aeruginosa gram-negative, bacilli and facultative aerobic, P. aeruginosa cause cystic fibrosis patients, wounds, burns, and immunodeficienct patients, that have many virulence factors such as pyocyanin , cytotoxic ,biofilm formation and motility, Eighty-eight isolates belonging to P. aeruginosa were collected including the 66 clinical isolates obtained from different hospitals in Baghdad and were from different sources and 22 environmental isolates from previous studies of soil near oil fields. Microscopical and cultural characteristics were studied and diagnosed using biochemical tests, VITEC device, their ability to adhere to non-living (Polystyrene), living cell line (A549) and cytotoxicity of bacterial filtrate
... Show MoreStaphylococcus aureus is a common pathogen associated with eye·s
infections. S. aureus is capable of biofilm fonnation, which increases its persistence and boots its levels of antimicrobial resistance . A total of 50
- aureus isolated from eyes <>f patientwith eye's infection : 41( 82%)
isolates were positive - alpha tox in production and 37 (74 %) isolates were posilive - biofilm formation .Where as 32 (64%) isolates were positive - alpha toxin production .and biotilm formation, 11 (22%) Lsolatcs were negative- alpha toxin production and biofilm formation and 7(14%) isolates were showed &nbs
... Show MoreCryptosporidiosis is an intestinal protozoan parasitic disease that infects human and animals, caused by apicomplexan parasite belong to the genusof Cryptosporidium. The current study was done to record the infection rate of cryptosporidiosis in human and cattle, and genotype the clinical isolates of Cryptosporidium in Baghdad Province. A total of 265 stool sample were collected (150 from human and 115 from cattle) during the period from December 2016 to the May 2017. Cryptosporidial infection was detected using modified acid fast stain. DNA of the parasite was extracted from oocysts of positive fecal samples and nested PCR method was used for partial 60 kDa glycoprotein (gp60) gene amplification then sequence analysis for selected samples.
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