The 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 pJET/bshA and pMG36e/bshA respectively. Both recombinants were transferred to E. coli MC1022 by chemical transformation. Obtained recombinants analyzed for expression and sequences. The results were confirmed that production of bile salt hydrolase from recombinant E. coli MC1022 pJET/bshA found to be higher while compared with E. coli MC1022 wild type and recombinant E. coli MC1022 pMG36e/bshA strain. However production of bile salt hydrolase from recombinant E. coli MC1022 pMG36e/bshA found to be higher while compared with E. coli MC1022 wild type. The recombinant plasmid also found to be stable in host organism after a few generations.
This research was aimed to the purification and characterization of cytosine deaminase as a medically important enzyme from locally isolated Escherichia coli; then studying its cytotoxic anticancer effects against colon cancer cell line. Cytosine deaminase was subjected to three purification steps including precipitation with 90% ammonium sulfate saturation, ion exchange chromatography on DEAE-cellulose column, and gel filtration chromatography throughout Sephadex G-200 column. Specific activity of the purified enzyme was increased up to 9 U/mg with 12.85 folds of purification and 30.85% enzyme recovery. Characterization study of purified enzyme revealed that the molecular weight of cytosine deaminase produced by E. coli was about 48 KDa,
... Show MoreReactive arthritis (ReA) has been as joint developing after infection, it belongs to spongylo arthritis (SpA). The etiology of this disease was multi factorial, the combination between genetic and environmental factors for triggering this disease. This study included 75 Iraqi Arab patients and 39 healthy control. Urine samples and blood were collected from each subject. The results showed that Escherichia coli bacteria (E. coli) was isolated from 32% of urine samples. HLA-B*27 allele frequencies was higher in ReA patients infected with E. coli. This lead to suggest that E. coli may be trigger factor in ReA patients with UTI which had HLA-B*27 positive.
A total of 165 clinical sample included Urine, Swab wounds and Burns were collected from Baghdad Governorate. Results showed that rate all isolates of E. coli was 50(30.3%) and rate of urine infection was 46(92%) and rate of swab wounds infection 4(8%). Where was diagnostic based on streaked on MacConkey agar, then single colony was transferred to Eosin Methylene Blue (EMB). Identification some of the biochemical test included: Catalase test, Oxidase test, Indole test, Methyl red, Vogues - Proskauer test and Citrate Utilization test. Then confirmed by the Vitek - 2 Compact System. The ability of E.coli isolate to biofilm formation to be studied it is considered one of the most important factors of virulence and has role in causing injury an
... Show MoreThree hospitals were chosen for the present (Maternity hospital, Raperin hospital and Rhizgari hospital) survey within Erbil city, 36 water samples were collected at regular monthly interval periods beginning at January to December 2012. Microbial analysis was done by selective medium and biochemical tests and the isolated bacteria from those hospitals were Eshcerichia coli, Acinetobacter lowffii, Klebsiella pneumoniae, Moraxilla spp., Salmonella Typhi, Citrbtobacter freundii, Vibrio fluvials, Acinetobacter haemolyticus, Weeksella zoohelcum, Pasteurella multicida, and Pseudomonas aeroginosa. E. coli isolates were subjected to antimicrobial susceptibility testing. In vitro activities of 10 different antibiotics against E. coli isolates we
... Show MoreThis study aimed to improve the microencapsulation technique using a type coating the encapsulation Layer by Layer, which provide the best protection for life Lactobacillus casei in the extrusion method and use the microencapsulation of materials of the protein concentrated by protein 80% and the coating with alginate and chitosan have the results showed the variation in the difference of the binding process encapsulation yield among the types of coating through. by studying of these the effect o stability of the bio probiotic free cell and the three types coated towards three different concentrations from bile salts 0, 0.3, 0.5 and 0.7% when the periods of time different of zero and two and three hours at incubation the recorded
... Show MoreIn present study 74 specimens of urine were collected from patients suffering from urinary tract infections.Fifty (67.56%) isolates were identified as Escherichia coli. 78% of isolates were identified as extendedspectrum beta lactamases (ESBL) producer. Antibiotic susceptibility t est was done and ceftazidime wasselected to complete this study by implying stress at sub-MIC on isolate harbor high number of resistancegenes (N11) and compared with sensitive isolate (S). Only four β-lactamase coding genes were detected;blaTEM, blaPER, blaVIM and blaCTX-M-2 and N11 had blaTEM, blaPER, and blaVIM. It was found that the resistantisolate did not form biofilm when compared with the sensitive one, which formed moderate biofilm. Inaddition, ceftazidi
... Show MoreThe detection for Single Escherichia Coli Bacteria has attracted great interest and in biology and physics applications. A nanostructured porous silicon (PS) is designed for rapid capture and detection of Escherichia coli bacteria inside the micropore. PS has attracted more attention due to its unique properties. Several works are concerning the properties of nanostructured porous silicon. In this study PS is fabricated by an electrochemical anodization process. The surface morphology of PS films has been studied by scanning electron microscope (SEM) and atomic force microscope (AFM). The structure of porous silicon was studied by energy-dispersive X-ray spectroscopy (EDX). Details of experimental methods and results are given and discussed
... Show MoreThe present study aims to detect the distribution of dfrA1 and cat1 antibiotic resistance genes among uropathogenic Escherichia coli (UPEC) in pregnant teen women and determine their susceptibility to common antibiotic uses. We collected urine (116) samples from patients in hospitals in Baghdad, Iraq. Isolation and identification of bacteria (culturing, biochemical test, and genetically by 16S rRNA gene), antibiotic susceptibility tests (eight antibiotics), and detection of the dfrA1 and cat1 resistance genes, and used SPSS program for statistically analyzing the results. The distributed UPEC in patients most than another causative agent in percentage (50%). It was highly resistan
... Show MoreBiofilm formation represents one of the biggest problems facing scientists because of this phenomenon linkage with virulence of bacteria and other clinical environmental problems. In the present study, two clinical isolates,
Escherichia coli, and Staphylococcus aureus were exposed to the non thermal plasma for different intervals of time (1, 2, 4, 8, and 16 min). The biofilm was measured post exposing. It was found that 2 min. exposing to non-thermal plasma reduced the biofilm formation by both clinical isolates significantly. It can be concluded that the ability of S. aureus to form biofilm higher than E. coli and exposing for 2 min to non-thermal plasma sufficient to reduce the biofilm formati